1.

INTRODUCTION

OBESITY :

Obesity is a chronic, multifactorial metabolic disorder characterized by the excessive

accumulation of body fat to an extent that adversely affects an individual’s health and functional
capacity. Clinically, obesity is often assessed using anthropometric indicators such as Body Mass
Index (BMI), where a BMI of 30 kg/m² or greater is categorized as obesity, reflecting a quantitative
measure of adiposity that correlates with health risk. However, reliance solely on BMI may
overlook critical aspects of fat distribution and composition, necessitating the use of
supplementary measures such as waist circumference and body fat percentage in both clinical and
research settings [1].

Over the past several decades, obesity has emerged as one of the most serious global public
health challenges of the 21st century, with prevalence rising at an alarming rate in both developed
and developing nations. This upsurge has been observed across all age groups, including children,
adolescents, and adults, underscoring the pervasive nature of the epidemic. The rapid increase in
obesity rates reflects profound shifts in lifestyles and environments that favor energy intake over
energy expenditure. In many populations, transitions toward diets high in energy-dense processed
foods, added sugars, and saturated fats, combined with reductions in physical activity due to
mechanization, urbanization, and sedentary occupations, have created environments that facilitate
excessive weight gain.

Importantly, the etiology of obesity extends beyond simple caloric imbalance and
encompasses a complex interplay of genetic, behavioural, environmental, socioeconomic, and
psychological factors. Genetic predisposition influences individual variability in appetite
regulation, metabolic efficiency, adipogenesis, and energy utilization, contributing to susceptibility
to weight gain. Behavioural factors, including dietary habits, eating patterns, and levels of physical
activity, interact with environmental determinants such as food availability, cultural norms, and
built environments that may limit opportunities for active lifestyles. Socioeconomic status further
shapes exposure to obesogenic influences, with disadvantaged populations often experiencing
higher risks due to limited access to healthy foods and safe recreational spaces. In parallel,
psychological factors, including stress, emotional distress, and sleep disturbances, influence
energy balance through effects on appetite, food choices, and metabolic processes [2].

The health consequences attributed to obesity are profound and wide-ranging. Obesity is
strongly associated with an increased risk of major non-communicable diseases (NCDs), including

1
cardiovascular disorders (such as coronary artery disease and stroke), type 2 diabetes mellitus,
hypertension, dyslipidaemia, and various malignancies, particularly cancers of the breast, colon,
and endometrium. The presence of obesity also exacerbates musculoskeletal disorders, contributes
to respiratory dysfunction, and diminishes overall quality of life. In addition to physical health
impacts, obesity is associated with significant psychosocial burden, including reduced self-esteem,
depression, social stigma, and impaired social functioning, which collectively amplify its burden
on individuals and communities.

Given the extensive health, social, and economic impacts of obesity, it constitutes a major
burden on healthcare systems worldwide. The chronicity and complexity of the condition
necessitate comprehensive strategies that integrate prevention, early detection, effective control,
and multidisciplinary management. Prevention efforts must address upstream determinants by
promoting healthy nutrition, increasing physical activity, and creating supportive environments
through public health policies and community-based interventions. Early diagnosis and routine
screening facilitate timely identification of individuals at risk, enabling intervention before the
onset of complications. Furthermore, effective control and management require individualized,
evidence-based approaches that encompass lifestyle modification, behavioural support,
pharmacotherapy when appropriate, and surgical interventions for selected cases of severe obesity.
Such integrative strategies are critical to mitigating the long-term health consequences and
economic burdens associated with the global obesity epidemic [3].

Fig. No 1: Obese

2
CAUSES OF OBESITY :

The etiology of obesity is complex and multifactorial, resulting from a sustained imbalance
between energy intake and energy expenditure. At its core, obesity develops when caloric
consumption consistently exceeds the body’s metabolic requirements, leading to excessive fat
accumulation. However, this imbalance is influenced by a wide range of interrelated biological,
behavioural, environmental, and socioeconomic determinants rather than a single causative
factor [1].

One of the primary contributors to obesity is unhealthy dietary behaviour. Modern dietary
patterns are increasingly characterized by high consumption of energy-dense, nutrient-poor foods
that are rich in saturated fats, added sugars, and refined carbohydrates. The widespread availability
of fast foods, sugar-sweetened beverages, and processed meals has significantly increased average
caloric intake while reducing dietary quality. Irregular meal patterns, large portion sizes, and
frequent snacking further exacerbate excessive energy consumption, thereby promoting weight
gain over time.

Fig. No 2: COMMON CAUSE OF OBESITY

3
 Physical inactivity is another major determinant of obesity. Advances in technology,
mechanization of labour, and increased reliance on motorized transportation have substantially
reduced daily energy expenditure. Sedentary behaviours, including prolonged screen time
associated with television viewing, computer use, and mobile devices, have become increasingly
prevalent across all age groups. Insufficient participation in regular physical activity diminishes
caloric utilization and contributes to the development of positive energy balance, thereby
increasing the risk of obesity [2].

Genetic predisposition plays a significant role in determining individual susceptibility to

obesity. Numerous studies have demonstrated that genetic factors influence appetite regulation,
satiety signalling, metabolic rate, fat distribution, and energy storage. Individuals with a family
history of obesity are more likely to develop the condition, particularly when exposed to
obesogenic environments. While genetic factors alone do not directly cause obesity, they interact
with environmental and behavioural influences to increase vulnerability to excessive weight gain.

Hormonal and metabolic abnormalities also contribute to the pathogenesis of obesity.

Endocrine disorders such as hypothyroidism, Cushing’s syndrome, polycystic ovary syndrome
(PCOS), and insulin resistance can disrupt normal metabolic processes, leading to reduced energy
expenditure and increased fat accumulation. Dysregulation of appetite-related hormones,
including leptin, ghrelin, and insulin, further impairs hunger control and satiety, promoting
overeating and weight gain [3].

Psychological and behavioural factors are increasingly recognized as important

contributors to obesity. Chronic stress, anxiety, and depression can influence eating behaviour,
often leading to emotional or stress-induced overeating. Sleep disturbances and insufficient sleep
duration have also been associated with hormonal changes that increase appetite and reduce energy
expenditure. These psychological factors not only contribute to the onset of obesity but also
complicate weight management efforts.

Environmental and socioeconomic factors play a critical role in shaping obesity risk at the
population level. Urbanization and modernization have created environments that promote
sedentary lifestyles and unhealthy dietary choices. Limited access to affordable, nutritious foods,
particularly in low-income communities, contributes to reliance on inexpensive, calorie-dense
alternatives. Additionally, the lack of safe and accessible spaces for physical activity reduces
opportunities for exercise, particularly among children and vulnerable populations [4]. These
broader social determinants interact with individual behaviours to perpetuate obesity across
generations [5].
4
DIAGNOSIS OF OBESITY:

The diagnosis of obesity is primarily based on anthropometric measurements. Body Mass

Index (BMI), calculated as weight in kilograms divided by height in meters squared (kg/m²), is the
most widely used screening tool. According to the World Health Organization (WHO), a BMI of
25.0–29.9 kg/m² is classified as overweight, while a BMI of 30.0 kg/m² or above indicates obesity
[6-8].

However, BMI does not distinguish between fat mass and lean body mass; therefore,
additional measures are often used for a more accurate assessment. These include waist
circumference, waist-to-hip ratio, and body fat percentage, which help assess central or abdominal
obesity. Advanced diagnostic techniques such as bioelectrical impedance analysis (BIA), dual-
energy X-ray absorptiometry (DEXA), and imaging methods may be employed in research and
clinical settings. Laboratory investigations are also conducted to identify obesity-related
comorbidities, including dyslipidaemia, hyperglycaemia, and hormonal abnormalities [6-10].

CONTROL AND MANAGEMENT OF OBESITY :

The control and management of obesity require a long-term, comprehensive, and

multidisciplinary approach that addresses the complex biological, behavioural, and environmental
factors underlying the condition. Given the chronic and relapsing nature of obesity, effective
management extends beyond short-term weight reduction and focuses on sustainable lifestyle
modification, behavioural change, and prevention of associated comorbidities. Interventions are
most effective when individualized and supported by coordinated efforts among healthcare
professionals, including physicians, dietitians, psychologists, and public health practitioners.

Dietary management constitutes a cornerstone of obesity control. It involves the adoption

of balanced, calorie-restricted dietary patterns designed to create a negative energy balance while
ensuring adequate nutrient intake. Diets emphasizing fruits, vegetables, whole grains, legumes,
lean protein sources, and healthy fats are recommended, alongside the reduction of saturated fats,
added sugars, refined carbohydrates, and ultra-processed foods. Portion control, mindful eating
practices, and regular meal timing further support weight reduction and maintenance. Long-term
adherence to healthy dietary habits is critical, as extreme or restrictive diets often result in weight
regain and metabolic disturbances.

Regular physical activity plays a pivotal role in obesity management by increasing energy
expenditure, preserving lean body mass, and improving metabolic and cardiovascular health.
Aerobic exercises such as walking, cycling, and swimming, combined with resistance training, are
5
recommended to enhance fat loss and improve muscle strength. Consistent engagement in physical
activity not only supports weight reduction but also improves insulin sensitivity, lipid profiles, and
overall physical and mental wellbeing. Reducing sedentary behaviours is equally important, as
prolonged inactivity contributes significantly to positive energy balance.

Behavioural interventions are essential for sustaining lifestyle changes and preventing
weight regain. Techniques such as self-monitoring of food intake and physical activity, goal
setting, problem-solving, and stress management enhance self-regulation and accountability.
Cognitive-behavioural therapy (CBT) has demonstrated effectiveness in addressing maladaptive
eating behaviours, emotional eating, and psychological barriers to weight control. Improving sleep
quality and addressing psychosocial stressors are also integral components of comprehensive
obesity management, as they influence appetite regulation and metabolic function.

Fig. No 3: RISK FACTORS FOR OBESITY

At the population level, public health measures play a crucial role in the control of obesity.
Community-based interventions, school and workplace wellness programs, and nutrition
education initiatives promote healthy behaviours and increase awareness of obesity-related risks.
Policy interventions aimed at improving food environments—such as regulating food marketing,
improving food labelling, and increasing access to affordable, nutritious foods—are essential for

6
creating supportive environments that facilitate healthy choices. Urban planning strategies that
promote active transportation and access to recreational spaces further contribute to population-
level obesity prevention and control [10,11].

TREATMENT OF OBESITY :

The treatment of obesity is individualized and depends on the severity of the condition, the
presence of obesity-related comorbidities, and patient-specific factors such as age, metabolic
status, psychological health, and motivation. Given that obesity is a chronic and relapsing disease,
treatment strategies aim not only at achieving weight reduction but also at maintaining long-term
weight loss and reducing associated health risks. A stepwise and comprehensive approach is
generally recommended, beginning with lifestyle modification and progressing to pharmacological
and surgical interventions when necessary.

Lifestyle interventions constitute the first-line treatment for obesity and form the
foundation of all weight management strategies. These interventions focus on dietary modification,
increased physical activity, and behavioural therapy to achieve a sustained negative energy
balance. Dietary interventions typically involve calorie restriction, improved diet quality, portion
control, and adherence to nutritionally balanced eating patterns. Regular physical activity is
encouraged to enhance energy expenditure, preserve lean body mass, and improve metabolic
health. Behavioural therapy supports these efforts by addressing maladaptive eating behaviours,
enhancing self-monitoring, goal setting, and coping strategies, thereby improving adherence to
lifestyle changes and reducing the likelihood of weight regain [12].

When lifestyle interventions alone fail to produce adequate or sustained weight loss,
pharmacological therapy may be considered as an adjunct treatment. Anti-obesity medications are
generally prescribed for individuals with a BMI ≥ 30 kg/m² or ≥ 27 kg/m² in the presence of
obesity-related comorbidities. These medications act through various mechanisms, including
appetite suppression, enhancement of satiety, reduction of fat absorption, or modulation of central
nervous system pathways involved in energy regulation. Pharmacotherapy has been shown to
produce modest but clinically meaningful weight loss and improvements in metabolic parameters
when combined with lifestyle modification. However, long-term use requires careful monitoring
due to potential side effects and variability in individual response [13].

In cases of severe obesity, particularly when conservative measures are ineffective,

bariatric surgery may be considered as a treatment option. Surgical intervention is generally
recommended for individuals with a BMI ≥ 40 kg/m² or ≥ 35 kg/m² with significant obesity-related

7
comorbidities such as type 2 diabetes mellitus, hypertension, or obstructive sleep apnoea. Common
bariatric procedures include Roux-en-Y gastric bypass, sleeve gastrectomy, and adjustable gastric
banding. These procedures promote weight loss through restrictive and/or malabsorptive
mechanisms and have been shown to result in substantial and sustained reductions in body weight.

Beyond weight reduction, bariatric surgery has demonstrated significant benefits in

improving or resolving obesity-related comorbidities, enhancing quality of life, and reducing long-
term mortality. However, surgical treatment requires careful patient selection, comprehensive
preoperative evaluation, and lifelong postoperative follow-up. Long-term success depends on
adherence to dietary recommendations, regular physical activity, nutritional supplementation, and
ongoing medical supervision to prevent complications and nutritional deficiencies [14,15].

Fig. No 4: Body Mass Index

8
 ➢ TYPES OF OBESITY :

Obesity can be classified into different types based on the distribution

of fat in the body and the age of onset.

• Generalized obesity - Excess fat is distributed throughout the body.

• Abdominal obesity - Excessive fat accumulates around the abdomen,
(visceral fat) Childhood obesity- Obesity that develops during
childhood [OR] Adolescence
• Adult obesity – Obesity that develops in adulthood

➢ BMI calculation :

The most commonly used diagnostic tool for obesity is the body
mass index [BMI] which is calculated by dividing a person’s weight in
kilogram by their height in meters square.

BMI = weight (kg) / (height (m))^2.

NORMAL WEIGHT -BMI greater than 18 to 24.9 kg\m2.

OVER WEIGHT - BMI greater than 25 to 29.9kg\m2.

OBESITY - BMI greater than 30 kg\m2.
OBESITY CLASS 1 - BMI of 30 to 34.9kg\m2.
OBESITY CLASS 2 - BMI of 35 to 39.9kg\m2.
OBESITY CLASS 3 – BMI greater than 40 kg\m2
(or)>35kg\m2 the presence of
comorbidities. (severe obesity)

Fig. No 5: BMI INDEX

9
ETIOLOGY AND RISK FACTORS :
• BEHAVIOUR :

• Physical inactivity
• Diet takeaway
• Cessation of smoking
• Eating patterns
a) Over eating
b) Night eating

c) Binge eating

• PATHOLOGICAL CONDITION :

• Hypothalamic obesity
• Decrease growth hormone
• Hypogonadism
• Hypothyroidism
• Polycystic ovarian syndrome
• Crushing syndrome (Female)

• MEDICATIONS :

• Glucocorticoids
• Antipsychotics
• Antidepressants
• Beta blockers
• Antidiabetics Insulin
• Sulphonylureas

COMPLICATIONS OF OBESITY :
• Stroke
• Stigma of obesity
• Obstructive sleep Apnoga
• Obesity hypoventilation syndrome
• Gastro Esophageal Reflux Disease (GERD)
• Heart disease & Hypertension
• Hyperlipidaemia

10
 • Fatty liver disease
• Diabetes
• Urinary incontinence

1. ADIPOCYTES :

Adipocytes or fat cells are specialized cells primarily responsible for storing energy in
the form of triacylglycerols (TAGS). It grows by increased cell number and size during in
utero life, childhood, and adolescence; after that, the number of adipocytes remains stable if
weight remains stable. In humans, adipose tissue appears between the 14th and 24th week of
gestation, Adipose tissue depots are highly dynamic and plastic organs. Changes resulting from
weight gain or loss

Fig. No 6: ADIPOST DIFFERENTIATION

The uptake of fatty acids, their conversion into TAGS & storage & the subsequent
release of fatty acids in energy is needed.
➢ ADIPOSE TISSUE :
• It is a connective tissue, but it’s also an interactive organ in your
endocrine system.
• That’s right, we’re talking about body fat.
• Adipose tissue communicates through hormone signals with other
organ throughout your body as well as with your central nervous
system to regulate your metabolism.

11
 ➢ TYPES OF ADIPOSE TISSUE :
✓ BROWN FAT CELL
✓ BEIGE FAT CELL
✓ WHITE FAT CELL

➢ DISTRIBUTION OF ADIPOSE TISSUE :

1. SUBCUTANEOUS ADIPOSE TISSUE:
This is the fat that lives between skin and muscles.
2. VISCERAL ADIPOSE TISSUE :
This is the fat that surrounds the organ in abdominal cavity.

OTHER LOCATION :

• Bone marrow
• Breast tissue
• Between muscles
• Around your heart
• Eye sockets
• Palms of your hand & soles of your feet

The Brown adipose tissue is mostly present in infancy and diminishes with age.
It’s found in upper back above clavicles and around vertebrae.

➢ ADIPOSE TISSUE TRANSCRIPTION FACTORS :

Adipose Tissue Abbreviation Agonists promote :

Transcription Factors
Peroxisome proliferator -activated PPARγ Increased adiposity, improved insulin sensitivity
receptor gamma (heterodimer with
RXR)
Peroxisome proliferator - activated PPARα Decreased serum triglycerides, LDL
receptor alpha cholesterol, increased HDL cholesterol,
protection against diet-induced obesity

12
 Peroxisome proliferator - activated PPARβ/δ Decreased LDL cholesterol, insulin, increased
receptor beta/delta HDL cholesterol, protection against diet-induced
obesity

Retinoid X receptor RXR Improved insulin sensitivity, reduced

(heterodimer with PPARγ) hyperglycemia, hypertriglyceridemia,
hyperinsulinemia, reduced weight gain and
food intake
Liver Transcription Factors
Liver X receptor LXR Decreased visceral adiposity, increased
subcutaneous adiposity, increased cholesterol,
increased plasma and hepatic triglycerides
Pregnane X receptor PXR Enhanced hepatic lipid accumulation, fatty
liver disease, increased hyperglycemia
Constitutive and rostane receptor CAR Improved insulin sensitivity, glucose and lipid
metabolism, protection against diet-induced
obesity, decreased glyuose levels
Farnesoid X receptor FXR Increased adiposity, improved insulin resistance.

SYSTEMIC HORMONE RECEPTORS AND THEIR ROLE IN OBESITY

AND OBESOGEN ACTION :

Hormone receptor Abbreviation Agonists promote :

Insulin receptor IR Increased adiposity
Estrogen receptors ER (α, β) Increased adiposity
Androgen receptor AR Decreased adiposity, lipid accumulation
Glucocorticoid receptor GR Increased adiposity
Thyroid hormone receptors TR (α, β) α- increased adiposity, increased cholesterol,
triglycerides β- reduction in cholesterol,
triglycerides, and adiposity

13
2. HORMONE :

➢ FLUENCE OF HORMONES IN OBESITY :

✓ Obesity involves a complex interplay of several hormones that

regulate appetite, metabolism and fat storage.
✓ Key hormones associated with obesity include.

• Leptin
• Adiponectin
• Ghrelin
• Insulin
• Growth hormone
• Sex hormone (estrogen, testosterone)

2.1. LEPTIN

A hormone produced by fat cells that signals to the brain to reduce appetite and
increase energy expenditure. In obesity, leptin resistance can occur, Where the brain
doesn’t respond as expected to leptin’s signals leading to continued hunger and reduced
energy expenditure.

➢ MECHANISM OF ACTION OF LEPTIN

Leptin receptors – leptin interacts with six type of receptors [o b - Ra- o b – R f, or

Lep Ra – Lep Rf ] which in turn are encoded by single gene LEPR. ob -rb is the
receptor isoform that can signal intracellularly and is present in hypothalamic nuclei
once leptin is bound to the ob-rb recptor. ob - rb: Regulating energy balance and body
weight by bind to the hormone leptin.
Stat 3 (Protein secondary messenger) – signal transducer & activator transcription 3,
role cell signaling and gene expression.

14
 Leptin bind to ob – rb receptor

It activate stat 3

Undergoes phosphorylation reaction

Travel to the nucleus

Modify the gene expression

Regulation of endocannabinoids

Increase appetite

2.2. ADIPONECTIN

➢ Adiponectin is another protein hormone a polypeptide consisting of 244

amino acids produced and secreted by adipose tissue cells (adipocytes)
➢ Adiponectin in humans is encoded by the ADIPOQ gene

➢ MECHANISM OF ACTION OF ADIPONECTIN

Adiponectin though produced by tissue cell (adipocytes)

It exerts its effects through brain

This is similar to the action of leptin, but the two hormones perform
complementary action and can have additive effect

15
2.3. GHRELIN

Ghrelin is a peptide hormone primarily produced in the stomach and sometimes

referred as the hunger hormone because it stimulates appetite and feeding behavior. It also
plays a role in energy homeostasis carbohydrate metabolism and growth hormone release
and produced by stomach, brain, small intestine, pancreas.

REGULATION OF GHRELIN LEVEL IN OBESITY

High level : The higher levels of ghrelin, there will be more the hungrier
Lower level : The the lower level, there will be the more full feeling

Weight lowering ghrelin can be beneficial for weight reduction

➢ FACTORS AFFECTING REGULATION OF GHRELIN HORMONE:

▪ Sleep

▪ Stress

▪ Exercise

MACHINSM OF ACTION OF GHRELIN

Ghrelin bind to GHS - R1a (Growth Hormone Secretagogue Receptors)

hypothalamus within the actuate

Cascade of Intracellular events

• G-PROTEIN
• Calcium
• Mitogen activated kinase
• Beta-arrest in recruitment

MACHINSM OF ACTION OF GHRELIN

➢ Intracellular events & Ghrelin

• PROTEIN ACTIVATION

Ghrelin binds to GHS-R1a, a G-protein coupled receptor, this binding

activates G-proteins, specially (Gaq/11) which are involved in intracellular signaling.
16
 • CALCIUM SIGNALING

The activation of Gaq /11 leads to stimulation of phospholipase C (PLC)

which hydrolyzes phospholitidylinositoe 4,5 phosphate (PIP2) into inositol triphosphate
(IP3) and diacylglycerol (DAG) IP3 then triggers the release of calcium ions (Ca2+)
from intracellular stores leading to changes in intracellular stores calcium levels.

MAKP PATHWAY ACTIVATION :

Ghrelin signaling can also activate the mitogen activated protein kinase
(MAPK) which plays a role in cell growth, proliferation, and differentiation.

BETA - ARRESTIN RECRUITMET :

Ghrelin signaling can also recruit beta-arrestin which is involved in

receptor internalization and desensitization, as well as other signaling cascades.

✓ Intracellular events influence the following physiological activities

• Neuronal activity
• Hormone secretion
• Affecting appetite
• Energy balance
• Physiological function

➢ ROLE OF GHRELIN HORMONE IN OBESITY

Increased level of ghrelin cause the obesity. Where the brain becomes less
responsive to ghrelin ( appetite-stimulating affects) potentially leading to food intake
and weight gain.

➢ MECHANISM OF ACTOIN OF GHRELIN IN OBESITY :

➢ Where the brain becomes less responsive to ghrelin’s appetite –

stimulating affects potentially leading to increase food intake and
weight gain
➢ Increased ghrelin stimulates the brain that stimulates growth
hormone secretion and food intake(appetite)
➢ It slows the metabolism and decrease the body’s ability to burn fat
and produce weight gain.

17
 Fig. No 7: MECHANISM OF ACTOIN OF GHRELIN

INSULIN :

Insulin is a polypeptide hormone consisting two peptide chain that are connected by
disulfide bonds. Insulin is secreted by β- cell of the pancreas. Insulin secretion is
regulated by blood sugar level certain aminoacids, other hormones and autonomic
mediators

INSULIN RESISTANCE AND OBESITY :

Insulin resistance is defined as the decreased tissue response to insulin-mediated cellular

actions. It has been proposed that insulin resistance results from reduced glucose transport,
particularly impairment of GLUT4, which may be due to increased reactive oxygen species (ROS)
from fatty acid oxidation via an unknown mechanism

18
19
ESTROGEN RELATED OBESITY :
• Promotes subcutaneous fat storage (hips,thighs,buttocks,- gynoid
pattern).
• Inhibits visceral fat accumulation (deep belly fat around organs).

MECHANISM OF ACTION OF ESTROGEN :

Estrogen binds to estrogen receptor (eRa ,eRβ) in fat tissue

Increases Lipoprotein lipase (LPL) activity in subcutaneous fat

Reduces the lipolysis (fat breakdown) in these areas.

Improves insulin sensitivity, which helps regulates glucose and

fat metabolism.
AFTER MENOPAUSE :
Oestrogen levels drop – shift in fat distribution to abdominal / visceral fat.
Associated with increased risk of metabolic syndrome, type 2 diabetes and
cardio vascular disease.

ESTROGEN IN MENOPAUSE RELATED IN OBESITY :

Menopause related obesity is linked to declining estrogen level

Leads to Shift in distribution and slows metabolism

Contributing to weight gain ( particularly around abdomen) visceral fat .

20
 Fig. No 8: ATURNING POINT IN METABOLIC HEALTH

PANCREATIC LIPASE :

Pancreatic lipase is a powerful lipolytic enzyme. It digests triglycerides into

monoglycerides and fatty acids. Activity of pancreatic lipase is accelerated in the
presence of bile. Optimum PH required for activity of this enzyme is 7 to 9.

FACTORS FOR DIGESTION OF FAT BY PANCREATIC LIPASE :

• Bile salts, which are responsible for the emulsification of fat, prior to their
digestion.
• Colipase, which is a coenzyme necessary for the pancreatic lipase to digest the
dietary lipids.
➢ About 80% of the fat is digested by pancreatic lipase.

➢ Deficiency (or) absence of this enzyme leads to excretion of undigested

fat in feces.

21
MECHANISM OF ACTOIN OF PANCREATIC LIPASE :
Enzyme Pancreatic lipase
Activator Alkaline medium
Acts on Triglycerides
End product Monoglycerides and fatty acids

PANCREATIC LIPASE CONTRIBUTES TO OBESITY :

❖ BREAKING DOWN DIETAEY FAT :
Pancreatic lipase converts the triglycerides into fatty acids and glycerol
facilitating fat absorption in the small intestine.

❖ INCREASE FAT ABSORPTION :

Efficient fat digestion and absorption can lead to increased intake,

Potentially contributing to weight gain and obesity.
❖ ENHANCING LIPID STORAGE :
Absorbed fatty acids can be stored in adipose tissue promoting fat
accumulation and obesity.
HIGH LEVEL :
High pancreatic lipase activity alone may not to be sole cause of obesity but it
can be a contributing factor.
LOW LEVEL:

Low levels of pancreatic lipase would likely lead to

1. Impaired fat digestion.
2. Reduced fat absorption.
ENDOCRINE DISTURBTING CHEMICALS (EDCS)

• The endocrine system is a network of glands of organs that produce store

and secrete hormones when functioning normally the endocrine system
works with others system to regulate bodys healthy development of
function throughout life.
• Endocrine disrupting chemicals are sub in the environment ( air ,soil ,or
water supply) food source personal care product and manufactured product
that interfere with the normal function of bodys endocrine system since
EDCS come from many different sources .When expose in the air as well
as the food and water.
22
 • EDCS also can enter the body through the skin by transfer from the body
through the skin and by transfer from mother to fetus (across the placenta)
or mother to infant ( via breast feeding) if a women has EDCS in her body
• They have been linked to human health issue related it alteration in sperm
quality and fertility abmormalities in sex organs endometrosis early
puberty alterd nervous system function immune function certain cancer
,respiratory problems ,metabolic issue ,diabetes, obesity cardiovascular
problem,growth neurological and learning disabilities and more.
• Eg: Bisphenol A (BPA), phthalates, Pestisides and Pollutant such as diorin
and polychlorinated biphenyls(PCBS)
ENDOCRINE DISTRUBTING CHEMICALS IN OBESITY :
EDCS can contribute to obesity by interfering with hormone function
particularly those involved in appetite regulations metabolism and fat storage.
EDCS can mimic block or alter the production and singnaling of hormones like leptin,
insulin, sex hormones leading to metabolic distruption and increase fat accumulation
Common EDCS implicated in obesity include bisphenol A ( BPA),phthalates
certain pesticides
❖ BISPHENOL (BPA) :
➢ In most cases BPA tend to accumulate in different tissue such as the
placenta, lungs kidney and liver . In these respective tissue beta
glucuronidase
enzyme exist in very high concentration

➢ As such this enzyme will deconjugate BPA and make it to exist

freely in its active forms.
➢ Since BPA naturally possesses lipophilic affinity it will bind to fats
in a high affinity further leading to its bioaccumulation and
promoting adipogenesis.
➢ Bisphenol A, a chemical found in many plastic of resins, has been
identified as a potential contribute to ability and related metabolic
disorders.
➢ It acts as endocrine disruptor, meaning it can interfere with body’s
hormone system including those that regulate metabolism and fat
storage.

23
 ❖ PHTHALATE :

➢ Phthalate are chemical found in plastic of personal care products

that can interfere with hormone regulation potentially contributing
to obesity.
➢ Commonly found in plastic have been linked to an increase risk of
obesity particularly in children and adults.
MECHANISM OF ACTION PHTHALATE :
❖ Phthalate exposure may contribute to weight gain by increase the number
and size of fat cells and disrupting body’s natural hormone balance,
including estrogen and progesterone.
❖ Phthalate are most widely used in various product:

✓ Food packaging: can leach from plastic packaging into food

✓ Personal care product: found in cosmetic, lotion and other personal

care item.

✓ Medical device: PH used in some medical devices.

✓ Building materials: present in plastic used in construction.

PESTICIDE :
Some pesticide as chlorpyrifos, pyrethroids and neonicotinoids may promote
obesity and other anthropometric changes by altering lipids and glucose
metabolism, modify genes or alter hormone levels such as leptin.

NEUROENDOCRINOLOGY OF OBESITY :

➢ Homeostatic mechanisms :
• Homeostatic feeding is defined as that which is required to meet
physiological/survival needs and is based on increasing the motivation to eat
when energy stores are depleted. In contrast, hedonic feeding is driven by
sensory perception or pleasure. The hedonic drive may override homeostatic
regulation even in the face of abundant energy stores via increased desire to
consume highly palatable foods.
• Regulation of energy balance, and hence weight status, relies on integrating
peripheral hunger and satiety signals by the central nervous system (CNS)
controlling feeding behavior and activity . Feeding behavior is often
dichotomized as homeostatic or hedonic

24
 ➢ Key CNS regions involved in the homeostatic pathway include the neuroendocrine
hypothalamus and the nucleus of the solitary tract (NST) in the lower brainstem.
There are two distinct populations of neurons within the arcuate nucleus (ARC) of
the hypothalamus. One population expresses the anorexigenic neuropeptide
precursor pro- opiomelanocortin (POMC), while the other population expresses the
orexigenic neuropeptides neuropeptide Y (NPY) and agouti-related protein (AgRP).
These neurons, in turn, project to other hypothalamic nuclei, including the
paraventricular nucleus (PVN), which integrate emotional and stress responses as
well as exert physiological control over metabolism via the release of thyrotropin-
releasing hormone (TRH) and corticotropin-releasing hormone (CRH) .
Serotonergic neurons from the dorsal raphe nucleus (DRN), which receive afferent
input from the spinal column and many parts of the brainstem, project to the ARC,
NTS and paraventricular nucleus (PVN). Depleting CNS serotonin results in
hyperphagia and obesity, while elevated CNS serotonin resulted in anorexia and
decreased energy intake insulin receptor and the leptin receptor recruit the low-
abundance-message insulin receptor substrate-2. Lack of available insulin receptor
substrate 2 for the leptin receptor due to hyperinsulinemia could result in defective
leptin signal transduction. Alternatively, insulin induction of suppressor of cytokine
signaling-3 could inactivate the leptin receptor through alterations in tyrosine
phosphorylation

HETEROGENEOUS NATURE OF CONTROL OF WEIGHT AND ADIPOSITY :

Genetics of obesity :
While genes certainly play a role in obesity pathogenesis, genetics alone cannot explain
the increase in obesity rates over the last 40 years, as genetics have not changed significantly
over that short time frame. Thus, the focus must turn to environmental factors as the cause
of the current obesity pandemic.

Environment and obesity :

Environmental factors that promote obesity include a host of alterations, including
nutrition, overeating and binge eating disorders/food addiction, time of eating, physical
inactivity, drugs, viruses, and the gut microbiome. Various obesogens exert their actions
within one or more of these pathways, and their actions will be discussed in the review
following this one. We recognize that there are other environmental factors involved in
obesity, such as the built environment, social disparities, and economic policies
25
 Nutrition and obesity :
Increased caloric intake correlates with weight gain but does not explain the mechanism
for the increased calorie intake. In addition, obesity has multifactorial origins and, nutrition is
more complicated than just counting calories. Indeed, all calories are not equivalent. An
attractive alternative hypothesis is that hyperinsulinemia is the primary factor driving energy
storage and weight gain. Indeed, the typical Western diet promotes obesity by increasing
calorically dense (high fat and high sugar) diets, which promote insulin secretion, insulin
resistance, and resultant weight gain. Furthermore, hyperinsulinemia interferes with leptin
signal transduction to increase food intake

Glycemic index vs. glycemic load :

A primary goal of improving metabolic health is to lower insulin levels. One way to do
so is to eat foods that produce a reduced postprandial glucose excursion (an indirect proxy for
insulin), such as with amylose. Higher glucose spikes upon eating are associated with more
inflammation and higher mortality . Some propose that a low-GI diet will keep blood glucose
down and help weight loss . However, the real reason that GI works is that it reduces insulin
spikes. Indeed, the glycemic load of carbohydrates consumed is a more important predictor
of weight gain than the actual number of calories.

Exercise and obesity :

Management of obesity is generally related to interventions in terms of lifestyle, mainly
through changes in diet directed at caloric restriction and adherence to exercise programs

Viruses and obesity :

Infection with seven different viruses and a scrapie agent has been associated with obesity
in animals and humans

PATHOPHYSIOLOGIC MECHANISMS THAT PROMOTE OBESITY :

➢ Insulin resistance and obesity

➢ Hepatic insulin resistance
➢ Brain insulin resistance
➢ Inflammation and obesity
➢ Gut microbiome and obesity
➢ Circadian rhythms and obesity

26
 2. LITERATURE REVIEW

➢ Yarmohammadi et al. (2021) reported that Azadirachta indica (neem) shows significant
protective effects against metabolic syndrome due to its anti-diabetic, anti-hyperlipidemic, anti-
obesity, and anti-hypertensive properties. These effects are mainly attributed to bioactive
compounds such as nimbidin, nimbolide, and flavonoids. Neem improves glucose metabolism
by enhancing insulin sensitivity and inhibiting α-glucosidase, while also regulating lipid levels
by reducing cholesterol and triglycerides. Additionally, its antioxidant and anti-inflammatory
activities help reduce oxidative stress and associated complications.

➢ Patil et al. (2024) investigated the inhibitory potential of Psidium guajava leaf extracts on
digestive enzymes involved in obesity. The study reported that guava leaves exhibit significant
inhibition of key enzymes such as α-amylase and pancreatic lipase, which are responsible for
carbohydrate and lipid digestion. This inhibition reduces nutrient absorption and may help in
controlling body weight and metabolic disorders and guava leaf extracts inhibit α-amylase, α-
glucosidase, and lipase, thereby reducing fat accumulation and postprandial glucose levels.

➢ Armstrong (2022) describes obesity as a complex, chronic disease influenced by genetic,

environmental, behavioral, and metabolic factors rather than merely a lifestyle issue. The
chapter highlights that obesity is associated with serious comorbidities such as diabetes,
cardiovascular diseases, and metabolic disorders. It emphasizes that effective management
requires a multidisciplinary approach, including dietary modification, physical activity,
behavioral therapy, pharmacological treatment, and in severe cases, bariatric surgery. It further
stresses that lifestyle interventions alone are often insufficient, and long-term management
strategies combining clinical and public health approaches are essential for sustainable weight
control

➢ Mukherjee et al. (2013) reported that several Indian medicinal plants exhibit a dual inhibitory
effect on digestive enzymes, particularly intestinal α-glucosidase and pancreatic lipase, which
play key roles in carbohydrate and lipid digestion. This combined inhibition is considered
highly effective for obesity management, as it reduces both glucose and fat absorption.

27
➢ Bhatt et al. (2017) reviewed various anti-obesity approaches and highlighted that obesity results
--primarily from an imbalance between energy intake and expenditure. The study emphasized
that conventional pharmacological treatments, though effective, often produce adverse side
effects, thereby necessitating safer alternatives such as herbal and natural products. The authors
also discussed mechanisms like appetite suppression, inhibition of fat absorption, and
enhancement of metabolism as key strategies in anti-obesity treatment.

➢ Choi et al. (2021) investigated the effects of Psidium guajava leaf extract on adipocyte
differentiation and insulin sensitivity using 3T3-L1 cell lines. The study demonstrated that the
extract significantly inhibited adipogenesis, as evidenced by a marked reduction in lipid
accumulation within adipocytes. This effect was primarily attributed to the downregulation of
key adipogenic transcription factors such as PPARγ, C/EBPα, and SREBP-1c, which play a
crucial role in the formation and maturation of fat cells.

➢ Mashitah et al. (2024) conducted a systematic review to evaluate the anti-obesity potential of
Cymbopogon citratus using evidence from in vitro, in vivo, and limited human studies. A total
of 18 studies (2003–2023) were analyzed. The findings indicate that different parts of the plant
(leaves, stalks, roots, and whole plant) exhibit anti-obesity activity due to the presence of
polyphenols, essential oils (citral), and dietary fiber. The study highlights multiple mechanisms
responsible for its anti-obesity effects, including inhibition of digestive enzymes, appetite
suppression, modulation of lipid metabolism, inhibition of adipogenesis, and increased energy
expenditure. Polyphenol-rich extracts primarily act by regulating lipid metabolism and
reducing fat accumulation, while essential oils enhance metabolic rate and reduce adipocyte
formation. Additionally, dietary fiber contributes by limiting fat absorption and improving lipid
profiles.

➢ Alhassan et al. (2025) evaluated the anti-obesity and cardioprotective effects of combined
polyphenols from Psidium guajava and Citrus limon leaves in a fructose-induced rat model.
The study reported significant improvement in metabolic parameters, including reduced lipid
accumulation and improved biochemical markers. The mechanism of action involves regulation
of lipid metabolism, oxidative stress reduction, and anti-inflammatory activity. The extract also
protected against fructose-induced cardiac injury by enhancing antioxidant defense systems.
Advanced analysis (LC-MS/MS) confirmed the presence of bioactive polyphenols responsible
for these effects.
28
➢ González-Molina et al. (2017) evaluated the anti-obesity potential of citrus-based bioactive
compounds, especially from lemon and related fruits. The study reported that citrus fruits are
rich in polyphenols, flavonoids (hesperidin, eriocitrin), and dietary fiber, which play a
significant role in metabolic regulation. The anti-obesity effects were attributed to mechanisms
such as enhanced lipid metabolism, increased β-oxidation, inhibition of adipogenesis, and
reduction of fat accumulation. Additionally, citrus compounds exhibited strong antioxidant and
anti-inflammatory properties, helping to reduce obesity-associated complications.

➢ Overdevest et al. (2018) investigated the effects of citrus flavonoid supplementation on exercise
performance in trained athletes using a randomized, double-blind study design. The study
demonstrated that daily intake of citrus flavonoid extract significantly improved power output
and exercise efficiency compared to placebo. The mechanism underlying this improvement is
attributed to the antioxidant properties of flavonoids and enhanced nitric oxide (NO)
production, which improve mitochondrial efficiency and oxygen utilization during exercise.
Additionally, supplementation reduced the oxygen consumption-to-power ratio, indicating
improved metabolic efficiency without affecting VO₂ max.

➢ Maji et al. (2020) reviewed the pharmacological importance of Azadirachta indica (neem)
leaves in the management of diabetes and obesity. The study highlights that neem leaves are
rich in bioactive compounds such as flavonoids, terpenoids, tannins, saponins, alkaloids, and
sterols, which contribute to their therapeutic effects. The anti-obesity activity of neem is mainly
attributed to its ability to inhibit pancreatic lipase and α-glucosidase enzymes, thereby reducing
fat absorption and carbohydrate digestion. Additionally, neem extracts were shown to improve
lipid profiles by reducing cholesterol, triglycerides, and LDL levels, indicating its role in
controlling obesity-related metabolic disorders.

➢ Patra et al. (2015) reviewed the role of various medicinal plants in the management of obesity
and highlighted the increasing global prevalence of obesity as a major health concern. The study
emphasized that obesity is primarily associated with disturbances in lipid metabolism, including
lipogenesis and lipolysis, and involves enzymes such as fatty acid synthase and lipoprotein
lipase. The review discussed that many synthetic anti-obesity drugs have limitations due to
adverse effects, leading to increased interest in herbal alternatives. Medicinal plants containing
polyphenols, flavonoids, alkaloids, and saponins were identified as effective in reducing body
weight and improving metabolic parameters
29
 ➢ BharatVed Research Team et al. (2021) described the therapeutic applications and dosage
regimens of various Ayurvedic formulations designed for chronic diseases affecting
neurological, renal, hormonal, and metabolic systems. The study highlights that these
formulations are composed of multiple herbal phytoconstituents with neuroprotective,
antioxidant, adaptogenic, and rejuvenating properties.

➢ BharatVed Research Team et al. (2021) described the therapeutic applications and dosage
regimens of various Ayurvedic formulations designed for chronic diseases affecting
neurological, renal, hormonal, and metabolic systems. The study highlights that these
formulations are composed of multiple herbal phytoconstituents with neuroprotective,
antioxidant, adaptogenic, and rejuvenating properties
.
➢ Saad et al. (2023) reviewed the anti-obesity potential of wild edible plants commonly included
in the Mediterranean diet and their bioactive compounds. The study highlights that obesity is
linked to energy imbalance and contributes to several chronic diseases such as diabetes,
cardiovascular disorders, and hypertension. The review emphasizes that plant-derived bioactive
compounds, including polyphenols, flavonoids, and dietary fibers, play a significant role in
obesity management. These compounds exert their effects through multiple mechanisms such
as appetite regulation, inhibition of pancreatic lipase, modulation of adipogenesis, enhancement
of thermogenesis, and stimulation of lipid metabolism and fat oxidation.

30
 3. AIM AND OBJECTIVES OF THE PRESENT STUDY

AIM :

The aim of the present study is to evaluate the in vitro Anti- obesity activity of
polyherbal leaves extracts from medicinal plants.

• Literature review of herbs for Anti -Obesity activity

• Collection of plants
• Authentication of plants
• Extraction of leaves
• Pharmacological activity
• Methodology (In vitro)
• Data analysis and interpretation

31
 4. PLAN OF WORK :

1. COLLECTION, PREPARATION OF PLANTS

2. AUTHENTICATION OF PLANT PLANTS
3. EXTRACTION OF PLANT MATERIAL
4. PHARMACOLOGICAL EVALUATION OF COMBINED LEAVE EXTRACTS OF SELECTED
PLANTS

ANTI- ANTI-OBESITY STUDIES

IN-VITRO ACTIVITY

• LIPASE-INHIBITORY ACTIVITY
• ACETYLCHOLINESTERASE INHIBITORY ACTIVITY
5. STATISTICAL ANALYSIS
6. RESULTS & DISCUSSION

32
 5. PLANTS PROFILE

DESCRIPTION
Neem (Azadirachta indica) is a highly valued evergreen tree belonging to the mahogany family
Meliaceae. Native to the Indian subcontinent, it thrives in tropical and semi-tropical climates and is
now cultivated in many parts of the world due to its exceptional medicinal and agricultural importance.
In traditional systems of medicine such as Ayurveda, Siddha, and Unani, neem is revered as a “sarva
roga nivarini” — a remedy for all diseases.

Fig. No 9 : Azadirachta indica

BOTANICAL CHARACTERISTICS

• Height: Neem trees typically grow 15–20 meters, but can reach up to 35 meters under
favorable conditions
• Leaves: The leaves are pinnate, bright green, and bitter, composed of multiple leaflets. They
are harvested for medicinal and cosmetic uses.
• Flowers: Small, white, and fragrant, arranged in drooping clusters. They bloom in spring
and are used in some traditional dishes and tonics.
• Fruit: A smooth, green drupe that turns yellow upon ripening, containing a seed from which
neem oil is extracted.
• Bark: Rough and grayish, with potent therapeutic compounds.

33
CHEMICAL COMPOSITION

Neem contains over 140 biologically active compounds. Key components include:
• Azadirachtin: A natural insect repellent, widely used in organic agriculture.
• Nimbin & Nimbidin: Known for anti-inflammatory and antimicrobial actions.
• Gedunin: Exhibits antifungal and antimalarial properties.
• Limonoids & Flavonoids: Support immune function and antioxidant activity.

MEDICINAL PROPERTIES

Neems known for a wide range of therapeutic effects:

• Antibacterial & Antiviral: Helps treat infections, skin ailments, and wounds.
• Antifungal: Effective against fungal infections such as ringworm.
• Anti-inflammatory: Reduces swelling, joint pain, and irritation.
• Immunomodulatory: Enhances the body's natural defenses.
• Detoxifying: Used in blood-purifying formulations.

TRADITIONAL USES IN HEALTHCARE

• Skin Care: Used to treat acne, eczema, psoriasis, and itching.

• Oral Health: Neem twigs are used as natural toothbrushes; extracts fight plaque and bacteria.
• Digestive Aid: Leaves and flowers are used in tonics to improve digestion.
• Diabetes Support: Traditionally used to help regulate blood sugar levels.
• Fever & Infection: Bark decoctions are used for reducing fever and fighting infections

PHYTOCONSTITUENTS PRESENT IN NEEM:

Category Examples in Neem Leaves

Limonoids (Triterpenoids) Azadirachtin, nimbolide, salannin, azadiradione,
gedunin
Flavonoids Quercetin, kaempferol
Alkaloids Various nitrogen compounds identified in extracts
Tannins Polyphenolic astringents
Saponins Foam-forming steroidal glycosides
Phenolics & Glycosides Plant antioxidants and attached sugar compounds

34
Phytochemical Constituents with % Content (Neem Leaf Extracts)

1. Phytochemicals in Ethanolic Leaf Extract (Quantified)

From a quantitative phytochemical screening of neem leaf extracts

Phytochemical Approx. % (Ethanolic extract)

Alkaloids ~1.38 %
Flavonoids ~0.73 %
Saponins ~0.42 %
Terpenes ~0.25 %
Tannins ~0.23 %
Steroids ~0.20 %
Phenols ~0.18 %
Cardiac Glycosides ~0.12 %
Anthraquinones ~0.03 %
Reducing Sugars ~0.12 %

Compound Group Potential Anti-obesity / Evidence Strength

Metabolic Role
Limonoids (e.g., May improve Moderate (mechanistic)
nimbolide) inflammation &
metabolism indirectly
Flavonoids (e.g., Known in other contexts Moderate (general literature)
quercetin) to assist fat oxidation &
insulin sensitivity
Enzyme inhibitors Can reduce fat/glucose Limited neem data but promising
(lipase/α- digestion
glucosidase)
Alkaloids / Minor antioxidant or Weak evidence specific to neem
Saponins / Tannins metabolic roles

35
LEMON DESCRIPTION :

• Kingdom: Plantae – the plant kingdom

• Clade: Angiosperms – flowering plants
• Clade: Eudicots – plants with two seed leaves and a specific pollen structure
• Clade: Rosids
• Order: Sapindales – an order that includes many aromatic plants and trees
• Family: Rutaceae – the rue or citrus family, which includes lemons, oranges, and other citrus
fruits
• Genus: Citrus – the genus of true citrus fruits
• Species: Citrus limon – the species name for the lemon

So, lemon leaves belong to the Rutaceae family and are characteristic of the Citrus genus, sharing traits
like aromatic oils and glossy foliage with other citrus plant.

Fig. No 10: Citrus limon

PHYTOCONSTITUENTS :

The chemical composition of lemon leaves is rich and complex, which is why they are valued in
traditional medicine, culinary uses, and aromatherapy. Here’s a detailed breakdown:

36
1. Essential Oils

Lemon leaves are aromatic due to volatile oils, which include:

• Citral – contributes to the lemony aroma
• Limonene – a monoterpene with antimicrobial properties
• Geraniol – has antioxidant and antimicrobial effects
• Linalool – offers a floral scent and calming effects

2. Phenolic Compounds

These compounds act as antioxidants and include:

• Flavonoids such as hesperidin, rutin, and quercetin
• Phenolic acids like caffeic acid and ferulic acid

3. Alkaloids

Some lemon leaves contain small amounts of alkaloids, which can have physiological effects, including
mild antimicrobial properties.

4. Tannins

Tannins contribute to astringency and have antioxidant and antimicrobial properties.

5. Vitamins and Minerals

Lemon leaves may contain trace amounts of:

• Vitamin C
• Calcium, potassium, and magnesium

6. Other Compounds

• Saponins – may contribute to anti-inflammatory and cholesterol-lowering effects

• Coumarins – have mild anticoagulant and antioxidant properties

Lemon Leaf Essential Oil – Major Chemical Components (% by GC-MS)

Compound Approx. Type / Notes

%
Limonene ~31.5% Major monoterpene hydrocarbon
Sabinene ~15.9% Monoterpene hydrocarbon
Citronellal ~11.6% Oxygenated monoterpene (aldehyde)

37
 Linalool ~4.6% Monoterpene alcohol
Neral (Z-citral) ~4.5% Oxygenated monoterpene (aldehyde)
Geranial (E-citral) ~4.5% Oxygenated monoterpene (aldehyde)
(E)-β-Ocimene ~3.9% Monoterpene hydrocarbon
Myrcene ~2.9% Monoterpene hydrocarbon
Citronellol ~2.3% Monoterpene alcohol
β-Caryophyllene ~1.7% Sesquiterpene hydrocarbon
Terpinen-4-ol ~1.4% Monoterpene alcohol
Geraniol ~1.3% Monoterpene alcohol
α-Pinene ~1.2% Monoterpene hydrocarbon
Other Minor Compounds ~1–3% Includes γ-terpinene, sylvestrene, and trace
total sesquiterpenes

1. Flavonoids

Flavonoids are the major bioactive compounds in lemon leaves with anti-obesity potential. Key
examples:

• Hesperidin – Reduces lipid accumulation, improves insulin sensitivity, and regulates

adipogenesis.
• Naringin – Suppresses fat formation, improves lipid metabolism, and exhibits antioxidant
activity.
• Rutin – Reduces oxidative stress associated with obesity and helps modulate fat metabolism.

2. Phenolic Acids

Phenolic acids in lemon leaves also contribute to anti-obesity activity:

• Caffeic acid – Inhibits adipocyte differentiation and fat accumulation.
• Ferulic acid – Reduces lipid levels and improves glucose metabolism.

3. Essential Oils / Terpenes

Volatile compounds in lemon leaves may indirectly support weight management:

• Limonene – Has anti-inflammatory and lipid-lowering effects, can reduce body fat in
experimental models.
• Citral and Geranial – May enhance energy expenditure and fat metabolism.

38
4. Alkaloids and Saponins

• Saponins – Reduce absorption of dietary fats and cholesterol in the intestines, aiding in weight
management.
• Alkaloids – Some alkaloids can stimulate metabolism and reduce fat accumulation.

Phytoconstituent Anti-obesity Mechanism

Hesperidin (Flavonoid) Inhibits fat cell formation, improves lipid metabolism
Naringin (Flavonoid) Reduces adipogenesis, regulates glucose/lipid balance
Rutin (Flavonoid) Antioxidant, modulates fat metabolism
Caffeic acid (Phenolic acid) Inhibits adipocyte differentiation
Ferulic acid (Phenolic acid) Improves lipid and glucose metabolism
Limonene (Terpene) Lipid-lowering, anti-inflammatory, promotes fat metabolism
Citral / Geranial (Terpenes) Enhances energy expenditure, fat oxidation
Saponins Reduce fat absorption in intestines

GUAVA DESCRIPTION :
The botanical classification of guava (Psidium guajava) is as follows:

• Kingdom: Plantae – the plant kingdom

• Clade: Angiosperms – flowering plants
• Clade: Eudicots – plants with two seed leaves
• Clade: Rosids
• Order: Myrtales – an order that includes aromatic plants like eucalyptus and clove
• Family: Myrtaceae – the myrtle family, which includes aromatic trees and shrubs
• Genus: Psidium – the guava genus
• Species: Psidium guajava – common guava

Fig. No 11: Psidium guajava

39
 Part Major Phytochemicals Function
Leaves Flavonoids: quercetin, Antioxidant, anti-
kaempferol, myricetin, inflammatory
rutin
Leaves Phenolic acids: gallic acid, Antioxidant, antimicrobial
ellagic acid, caffeic acid
Leaves Tannins: catechins, Astringent, antimicrobial
ellagitannins
Leaves Terpenes: caryophyllene, Antimicrobial, anti-
limonene, eucalyptol inflammatory
Fruit Vitamins: C, A, folate Immunity, antioxidant
Fruit Flavonoids & phenolics Antioxidant, anti-
inflammatory
Fruit Carotenoids: lycopene Antioxidant,
cardioprotective
Fruit Organic acids: citric, malic Flavor, digestion

1. Flavonoids

Flavonoids are the most important anti-obesity compounds in lemon leaves. Key examples:
Flavonoid Anti-obesity Mechanism
Hesperidin Reduces adipogenesis (fat cell
formation), improves lipid metabolism,
and regulates insulin sensitivity.
Naringin Suppresses fat accumulation, modulates
lipid metabolism, and has antioxidant
activity.
Rutin Reduces oxidative stress linked to
obesity and modulates adipocyte
differentiation.

40
2. Phenolic Acids

Phenolic acids in lemon leaves contribute to anti-obesity activity :

Compound Mechanism
Caffeic acid Inhibits adipocyte
differentiation and lipid
accumulation.
Ferulic acid Reduces triglyceride levels and
improves glucose metabolism.

3. Terpenes / Essential Oil Components

Volatile compounds in lemon leaves may also help manage obesity indirectly:

Compound Mechanism
Limonene Reduces body fat and has anti-
inflammatory effects.
Citral / Geranial Promote energy expenditure and
lipid metabolism.

4. Saponins

• Saponins reduce fat absorption in the intestines and help in weight management.

Phytoconstituent Anti-obesity Effect / Mechanism

Hesperidin (Flavonoid) Inhibits fat cell formation, improves

lipid metabolism

Naringin (Flavonoid) Suppresses adipogenesis, regulates

glucose/lipid balance

Rutin (Flavonoid) Antioxidant, reduces oxidative stress

in obesity

41
Caffeic acid (Phenolic) Inhibits adipocyte differentiation

Ferulic acid (Phenolic) Reduces triglycerides, improves

glucose metabolism

Limonene (Terpene) Promotes fat metabolism, anti-

inflammatory

Citral / Geranial (Terpenes) Increase energy expenditure and lipid

oxidation

Saponins Reduce fat absorption in intestines

42
 6. MATERIALS AND METHODS

COLLECTION, PREPARATION AND AUTHENTICATION OF PLANT MATERIALS

The whole plants of the selected herbs was collected in selected area around Tiruvallur district.
The plants were authenticated by Siddha Central Research Institute, Anna Hospital campus,
Arumbakkam, Chennai, Tamil Nadu. The voucher specimens of the plants are maintained at the
herbarium. Morphological and molecular methods will be employed to authenticate the plant.

1. Preparation of Leaf Powder from Neem, Lemon, and Guava Leaves

The process begins with the collection of fresh leaves from neem, lemon, and guava
plants. Only healthy, mature, and disease-free leaves are selected to ensure the quality of the
final product. Immediately after collection, the leaves are transported to the processing area in
clean containers to avoid contamination.

i. Cleaning and Sorting

The collected leaves are thoroughly washed using clean water to remove dust,
dirt, insects, and other foreign materials. In some cases, a mild disinfectant solution
may be used to eliminate microbial contaminants. After washing, excess surface water
is drained, and the leaves are manually sorted to remove damaged, discolored, or
infected leaves. The leaves are then separated from stems and other unwanted plant
parts. Approximately 3 kg of cleaned leaves are taken for further processing.
ii. Drying Process
The cleaned leaves are spread evenly in a thin layer on drying trays to ensure
proper air circulation and uniform drying. Drying is a crucial step as it reduces moisture
content and prevents microbial growth while preserving the bioactive compounds.
iii. Natural Drying

The drying process is continued until the leaves reach a constant weight, indicating
that most of the moisture has been removed. Initially, fresh leaves contain about 70%
moisture, which is reduced to approximately 4–5%, making them suitable for long-term
storage and processing.

43
 iv. Grinding and Size Reduction
Once dried, the leaves become brittle and are ready for size reduction. They are fed
into a pulverizer or grinding machine, where they are converted into fine powder. Care
is taken to avoid overheating during grinding, as excessive heat can degrade sensitive
bioactive compounds.
v. Sieving

The powdered material is passed through a sieve or mesh screen to obtain a uniform
particle size. This step ensures consistency in texture and improves the quality of the final
product, making it suitable for further applications such as herbal formulations or
analytical studies.

2. Weighing, Proportioning, and Blending of Polyherbal Formulation

Accurate weighing and proper blending of powdered ingredients are essential steps in
the preparation of a polyherbal formulation, as they directly influence the uniformity, quality,
and therapeutic efficacy of the final product.

i. Weighing and Proportioning

Each of the prepared plant leaf powders, namely neem (Azadirachta indica), lemon
(Citrus limon), and guava (Psidium guajava), was accurately weighed using a calibrated digital
analytical balance. Prior to weighing, the balance was standardized to eliminate any zero error,
ensuring precision in measurement. Clean and dry weighing boats were used to prevent
contamination and loss of material.

The quantities of individual powders were determined based on the formulation design.
In the present study, the powders were mixed in a ratio of 1:1:1, wherein equal quantities of
each plant material were taken. For example, 10 g of neem leaf powder, 10 g of lemon leaf
powder, and 10 g of guava leaf powder were weighed separately. The weighed samples were
properly labeled to avoid any mix-up during subsequent processing steps.

ii. Blending and Mixing

The accurately weighed powders were transferred into a clean, dry, and inert mixing
container made of stainless steel. Care was taken to ensure that the container was free from
moisture, dust, and any residual substances from previous operations.

44
 Blending of the powders was carried out to achieve a homogeneous mixture using
appropriate mixing techniques. Depending on the scale of preparation, both mechanical and
manual methods were employed.

3. Methodology:
i. Soxhlet extraction of plant powder:

The 10 g of Guava powder, 10 g of Lemon powder, and 10 g of Neem powder

samples were subjected to Soxhlet extraction (Borosil) using Distilled Water as solvent.
The powdered samples were made a 30 g thimble using handmade filter paper. The plant
sample filled thimble was carefully placed inside the extractor chamber and poured with
selected solvent ethanol or Distilled Water as 1:10 ratio. The reservoir round bottom flask
was heated to 55-60°C in a heating mantle. At least 20 refluxes were run to get good quality
plant solvent extract. The resultant solvent extract was condensed using a rotary evaporator
(Buchi, Bangalore, India) under reduced temperature in vacuum condition. The resultant
extract was collected in a container for further analysis and storage under -20°C.

Fig. No 12: SOXHLET APPARATUS

45
Lipase- Inhibitory activity :

Lipase inhibitory activity of samples was determined using a modified assay method
[Etoundi et al., 2010]. Briefly, a suspension containing 1% (v/v) triolein and 1% (v/v) Tween
40 in 0.1 M phosphate buffer (pH 8) was prepared and emulsified. Assay was then initiated by
adding 1600 μl of the triolein emulsion to 200 μl of porcine pancreatic lipase (0.5 gm pancreatin
in 15ml of 0.1 M phosphate buffer at pH 8.0) and 200μl of sample at different concentrations.
The test tubes containing reaction mixture were incubated at 37°C for 30 min and then the
absorbance was recorded at 450 nm and designated as T30. The % inhibition was calculated
using the following formula.

A450 Control - A450 Extract

% inhibition = -------------------------------------- × 100

A450 Control

Determination of Acetylcholinesterase Inhibitory Activity

• The test sample extract was screened for their inhibitory activity against AChE enzyme using
a modified Ellman’s method.

• Add 200 µl of plant extract of different concentrations 25 µg/ml, 50 µg/ml, 100 µg/ml,
250µg/ml, and 500µg/ml in a test tubes respectively with 500𝜇L of DTNB solution, 100 𝜇L
of enzyme (AChE, 2U/mL), 1000𝜇L 50 mM Tris–HCl buffer.

• For control tube, add everything except plant extract. Instead 200 µl of Dis. water was added.

• After mixing, the tubes were incubated for 15 min (37ºC) and then the absorbance was
measured at 412 nm and the readings were used as blank.

• The enzymatic reaction was initiated by the addition of 150 µl of ATCI.

• Total mixtures were incubated for a further 5 minutes at 28˚C. or Then the hydrolysis of
acetylthiocholine was monitored by reading the absorbance every 5 min for 20 min.

• The percentage inhibition was calculated as follows: %Inhibition= (E-S)/ E*100

• Where; E is the activity of the enzyme without extract and S is the activity of enzyme with
the extract. IC50 value could be calculated from the % inhibition values of different
concentrations of each plant extract.

46
 7. RESULTS AND DISCUSSION

[Link] Yield

[Link]. Sample (Grams) Solvent ml Obtained (gram) Yield %

1 30 Distilled water- 300 2.959 9.86

[Link]- Inhibitory activity

Concentration Absorbance Average Inhibition

(µg/mL) I II III (%)
Control 1.396 1.389 1.324 1.370 0.000

31.25 1.206 1.211 1.207 1.208 11.813

62.5 1.166 1.161 1.161 1.162 15.130

125 1.105 1.102 1.104 1.104 19.433

250 1.085 1.091 1.090 1.088 20.548

500 0.790 0.798 0.750 0.779 43.122

Orlistat- 200 0.253 0.253 0.254 0.253 81.504

Interpretation:

i. The test extract demonstrated a clear concentration-dependent inhibition of lipase activity over
the range of 31.25–500 µg/mL. The percentage inhibition increased from 11.81% at 31.25
µg/mL to 43.12% at 500 µg/mL, indicating a moderate and dose-responsive inhibitory effect.

ii. Notably, a substantial increase in inhibition at higher concentration (500 µg/mL) suggests
improved efficacy at elevated doses, though the extract did not achieve 50% inhibition within
the tested range.

47
iii. In comparison, the standard drug Orlistat exhibited 81.50% inhibition at 200 µg/mL,
confirming the robustness of the assay and demonstrating significantly higher potency than the
test extract.

iv. The calculated IC₅₀ value (639.35 µg/mL) indicates moderate inhibitory activity, as the extract
requires relatively higher concentrations to achieve 50% inhibition compared to potent lipase
inhibitors.

[Link] of Acetylcholinesterase Inhibitory Activity

Before adding substrate

Concentration (µg/ml) OD-1 OD-2 OD-3 Average

25µg/ml 0.287 0.275 0.266 0.276

50µg/ml 0.292 0.279 0.266 0.279
100µg/ml 0.283 0.279 0.277 0.280
250µg/ml 0.280 0.276 0.265 0.273
500 µg/ml 0.273 0.271 0.281 0.275
Control 0.289 0.273 0.278 0.280

Absorbance after 5 min of adding substrate

Concentration (µg/ml) OD-1 OD-2 OD-3 Average
25µg/ml 1.523 1.520 1.514 1.519
50µg/ml 1.315 1.319 1.315 1.316
100µg/ml 1.101 1.103 1.100 1.101
250µg/ml 1.062 1.089 1.029 1.060
500 µg/ml 1.022 1.019 1.029 1.023
Control 0.525 0.545 0.549 0.540

Absorbance after 10 min of adding substrate

25µg/ml 1.517 1.512 1.509 1.513
50µg/ml 1.261 1.261 1.261 1.261
100µg/ml 1.034 1.039 1.008 1.027
250µg/ml 1.026 1.017 0.986 1.010

48
500 µg/ml 1.006 1.017 0.986 1.003
control 0.626 0.668 0.643 0.646

Absorbance after 15 min of adding substrate

25µg/ml 1.478 1.477 1.477 1.478

50µg/ml 1.207 1.207 1.207 1.207

100µg/ml 0.966 0.976 0.970 0.970
250µg/ml 0.966 0.963 0.964 0.964
500 µg/ml 0.936 0.934 0.941 0.937
control 0.692 0.692 0.692 0.692

Absorbance after 20 min of adding substrate

25µg/ml 1.307 1.305 1.306 1.306
50µg/ml 1.156 1.165 1.154 1.158
100µg/ml 0.942 0.950 0.964 0.952
250µg/ml 0.935 0.932 0.932 0.933
500 µg/ml 0.906 0.903 0.904 0.904
control 0.762 0.762 0.786 0.770

Consolidated results

Concentration %
(µg/ml) inhibition

25µg/ml 8.756

50µg/ml 28.509

100µg/ml 31.654

250µg/ml 42.446

500 µg/ml 42.801

49
Fig. No 13: DILUTION FOR LIPASE INHIBITION

50
 Fig. No 14: PERCENTAGE INHIBITION OF ACETYLCHOLINESTERASE

Fig. No 15: LIPASE

DISCUSSION
51
• The inhibitory activity of the test extract against Acetylcholinesterase was evaluated at
different concentrations (25–500 µg/mL). The percentage inhibition increased with
increasing concentration of the extract, indicating a concentration-dependent inhibitory
effect.

• At the lowest tested concentration (25 µg/mL), the extract exhibited 8.76% inhibition,
suggesting minimal inhibition of enzyme activity. As the concentration increased to 50
µg/mL, the inhibition increased significantly to 28.51%, indicating moderate enzyme
inhibition.

• Further increases in concentration produced a gradual rise in inhibitory activity, with

31.65% inhibition at 100 µg/mL and 42.45% inhibition at 250 µg/mL. At the highest
tested concentration (500 µg/mL), the extract showed 42.80% inhibition, which is only
slightly higher than the inhibition observed at 250 µg/mL.

• This pattern suggests that the inhibitory activity begins to plateau at higher
concentrations, indicating that maximum inhibition may be approaching within the
tested range.

• The IC₅₀ value of 503.664 µg/mL was estimated by extrapolation of the concentration–
response curve, as the maximum inhibition observed within the tested concentration
range (500 µg/mL) was 42.8%, which is below the 50% inhibition threshold.

52
 8. CONCLUSION

The result of the present study contributes the evaluation of combined leave extract of
Azadirachta indica, Citrus limon and Psidium guajava for Anti-Obesity effects by in-vitro methods

The test extract exhibits a dose-dependent and moderate lipase inhibitory activity, with
appreciable inhibition observed at higher concentrations. However, its efficacy remains lower than the
standard, indicating limited but notable anti-lipase potential under the present study conditions.

The extract demonstrates the potential acetylcholinesterase inhibitory activity, but higher
concentrations or further purification may be required to achieve stronger inhibitory effects.

From the result, It is indicated that combined extracts of Azadirachta indica, Citrus limon and
Psidium guajava possess the Anti-Obesity effect and safer alternate for treatment of obesity.

In future further investigation might an insight to identify and characterization of exact active
phytoconstituent responsible for Anti-Obesity by net work pharmacology and to elucidate the
mechanism of action which is responsible for the observed significant activity an inhibition lipase and
Acetylcholinesterase enzyme in compare to standard drugs (Orlistat)

53
 REFERENCE

1. Haslam D.W., James W.P. "Obesity". Lancet. 2005; 366: 1197-209.

2. Kushner R.F., Bessesen D.H. Treatment of the obese patient. Endocrinolog
2009 158: 916929.

3. Gupta 5.K, Khandelwal M., Pancholi 5.5, Oupta M.K., Khinchi M.P.
Obesity- Challenges and treatment: a review, LJRAP. 2010; 1(2): 350-357

4. Adams J.P., Murphy P.G. "Obesity in anaesthesia and intensive care". Br J

Anaesth 2000: 8591-108.

5. Imaz 1., Martines-Cervell C Garcia-Alvarez EE Sendra-Gutierres J.M.,

Gonaalea Enriques J. "Safety and effectiveness of the intragastric balloon
for obesity. A meta-analysis, Obes Surg 2008, 18: 841-6.

6. Shah Y.R., Sen D.J., Patel R.N., Patel J.S., Patel A.D., Prajapati P.M.
Liposuction: A Remedy From Obesity. Int. J. Drug Dev. Res. 2011; 3(1): 14-
30.

7. Grill H., Ginsberg A., Seeley R., Kaplan J. Brainstem application of

melanocortin receptor ligands produces long-lasting effects on feeding and
body weight. J Neurosci., 1998; 18: 10128-35.

8. Pusku P., Stahl T. Health in all policies-the Finnish initiative background,

principles and curren33333t issues. Annu Res Public health, 2010; 31: 315-
328.

9. Woods S., Latter K., McKay L., Porte D. Chronic intracerebraventricular

infusion of insulin reduces food intake and body weight of ba boons. Nature.
1979, 282: 503-5

10. Zhang Y., Proenca R., Mallei M., Barune M., Leupold L, Friedman J.M.
Positional cloning of the mouse obese gene and its human homologue.
Nature. 1994, 372: 425-32

11. Diament AL., Fisler J.S., Warden C.H. Obesity alleles in mice and humans.
Obesity Reviews 2003: 4: 249-255

54
12. Sweeting H.N. Measurement and definitions of obesity in childhood and
adolescence: A field guide for the uninitiated. Nutr J.2007, 6:32
13. Gray D.S., Fujinka K. Use of relative weight and Body Mass Index for the
determination of adiposity. J Clin Epidemiol 1991; 44:545-50
14. Flegal K.M., Ogden CL, Wei R., Kocamarski R.L, Johnson C.L. "Prevalence
of overweight in US children: comparison of US growth charts from the
Centers for Disease Control and Prevention with other reference values for
body mass index. Am. J. Clin. Nutr 2001: 73:1086-
15. Basión D.G., Wilcox H., Figlewica D.P., Dorsa D.M. Insulin and insulin-like
growth factors in the CNS Trends Neurosci. 1988:11:107-11
16. Sturm R. "Increases in morbid obesity in the USA" Public Health 2005; 121:
4926
17. World Health Organization. (2025). Obesity and overweight. Retrieved from
[Link]
18. Centers for Disease Control and Prevention. (2025). Adult Obesity Causes
& Consequences. Retrieved from
[Link]
19. Ng, M., Fleming, T., Robinson, M., et al. (2014). Global, regional, and national prevalence of
overweight and obesity in children and adults during 1980–2013: a systematic analysis. The
Lancet, 384(9945), 766–781.
20. Hill, J. O., Wyatt, H. R., & Peters, J. C. (2012). Energy balance and obesity. Circulation,
126(1), 126–132.
21. Bray, G. A., Kim, K. K., & Wilding, J. P. H. (2017). Obesity: A chronic relapsing progressive
disease process. Obesity Reviews, 18(7), 715–723.
22. World Health Organization. (2000). Obesity: Preventing and managing the global epidemic
(WHO Technical Report Series No. 894). World Health Organization.
23. National Institutes of Health. (1998). Clinical guidelines on the identification, evaluation, and
treatment of overweight and obesity in adults. NIH Publication No. 98-4083.
24. Kyle, U. G., Bosaeus, I., De Lorenzo, A. D., et al. (2004). Bioelectrical impedance analysis—
part I: review of principles and methods. Clinical Nutrition, 23(5), 1226–1243.
25. Rothman, K. J. (2008). BMI-related errors in the measurement of obesity. International
Journal of Obesity, 32(S3), S56–S59.
26. Jensen, M. D., Ryan, D. H., Apovian, C. M., et al. (2014). 2013 AHA/ACC/TOS guideline for
the management of overweight and obesity in adults. Circulation, 129(25), S102–S138.

55
27. Knowler, W. C., Barrett-Connor, E., Fowler, S. E., et al. (2002). Reduction in the incidence of
type 2 diabetes with lifestyle intervention. New England Journal of Medicine, 346(6),
393–403.
28. Wing, R. R., & helan, S. (2005). Long-term weight loss maintenance. American Journal of
29. Clinical Nutrition, 82(1), 222S–225S.
30. Wilding, J. P. H., Batterham, R. L., Calanna, S., et al. (2021). Once-weekly semaglutide in
adults with overweight or obesity. New England Journal of Medicine, 384(11), 989–1002.
31. Buchwald, H., Avidor, Y., Braunwald, E., et al. (2004). Bariatric surgery: A systematic
review and meta-analysis. JAMA, 292(14), 1724–1737.

56