INTRODUCTION:

Obesity is defined by abnormal accumulation of body fat in the subcutaneous and visceral areas, posing significant health risks¹. Its tight relationship to chronic disorders including Type II diabetes and cardiovascular diseases has resulted in an alarming rise in prevalence over the past few decades, placing a significant strain on global healthcare systems. Genetic, environmental, and behavioural variables interact in a complicated way to cause obesity. The main causes of this expanding problem are sedentary lifestyles and high-calorie consumption. While available, the current pharmacological medications for obesity and hyperlipidemia have lessened the burden, underscoring the need for more efficacious interventions^2,3.

Studies using rat models have demonstrated that high-fat diets significantly alter lipid profiles and induce insulin resistance, contributing to metabolic dysfunction. For example, obesity induced by high-fat diets in rats has been associated with increased serum fatty acids, leading to lipotoxicity in various organs⁴. This underscores the intricate connection between dietary fat, lipid metabolism, and the development of obesity-related metabolic disorders⁵. Ficus benjamina, popularly recognized as weeping fig, is a tropical plant from the Moraceae family, valued for both ornamental and medicinal uses. Native to Southeast Asia and Australia, it contains bioactive compounds such as flavonoids, alkaloids, polyphenols, triterpenoids, and tannins^6,7. These constituents contribute to its pharmacological properties, including anti-inflammatory, antioxidant, antidiabetic, and lipid-lowering effects. Notably, Ficus benjamina has shown potential in managing metabolic disorders like obesity by improving lipid profiles and reducing oxidative stress ^8,9.

The current study investigates the hydro ethanolic extract of Ficus benjamina for its potential ameliorative effect on obesity markers in the rats subjected to a high-fat diet. This research aims to evaluate efficacy of Ficus benjamina as a therapeutic agent for managing obesity and associated metabolic disorders.

MATERIALS AND METHODOLOGY:

Acquisition and extraction of botanical specimens:

Leaves of Ficus benjamina were collected from the Botanical Garden, Sarangpur, Chandigarh, and authenticated by NISCAIR, New Delhi (Ref: NISCAIR/RHMD/consult/2021/3828-29-1). A total of 1.4 kg of powdered leaves was extracted using 80:20 ethanol-water. The extract was concentrated under reduced pressure at 50–60°C using a rotary evaporator, yielding 26.34g of crude extract, labeled as F1 and stored appropriately¹⁰.

Fractionation of Hydroalcoholic Fraction (F1):

The hydroalcoholic extract (F1) of Ficus benjamina was fractionated using sequential solvent extraction with solvents of increasing polarity: n-hexane (F2), ethyl acetate (F3), acetone (F4), and ethanol (F5). F1 was dissolved in ethanol and partitioned using a separating funnel. Each fraction was concentrated and subjected to phytochemical screening to identify secondary metabolites using standard protocols¹¹.

Determination of Phytoconstituents by using HPTLC:

HPTLC (High-Performance Thin-Layer Chromatography) was utilized to quantify phytoconstituents in the hydroalcoholic extract (F1) of Ficus benjamina. Silica Gel 60 F254 plates were utilized for chromatographic separation during the analysis, after pre-activation of the plates using a Linomat-V applicator under nitrogen flow. Plates were prewashed with methanol and activated at 60°C. Sample bands (6 mm) were applied 8 mm from the base at 150nL/s. Chromatographic development used a linear ascending method in a twin chamber saturated with mobile phases: ethyl acetate: toluene: formic acid (3.5:5:0.5) for rutin and toluene: chloroform: methanol (7:2.5:0.5) for quercetin. Densitometric scanning was conducted at 254 nm (quercetin) and 366 nm (rutin) with a CAMAG TLC scanner III^12,13.

Assay for the Inhibitory Effect on Pancreatic Lipase Inhibition:

Lipase activity was assessed through the detection of p-nitrophenol release from p-nitrophenyl palmitate at 405 nm using a microplate reader. Porcine pancreatic lipase (1mg/mL) and plant extracts were prepared in Tris-HCl buffer (pH 8.5), with PNPP diluted in acetonitrile: ethanol (1:2)¹⁴. The reaction mixture (200µL) contained 0.1mg/mL lipase, 0.167mM PNPP, and varying concentrations (5–200µg/mL) of Ficus benjamina extract and its fractions (F1–F5), with orlistat as a positive control. Reactions were incubated at 30°C, and lipase inhibition was measured in triplicate. IC₅₀ values were calculated to determine the extract concentration required for 50% inhibition^15,16,17.

EXPERIMENTAL ANIMALS:

The study employed male Wistar rats (7–8 weeks old), which are acquired from the animal facility of Chandigarh College of Pharmacy, Landran. Male Wistar rats were housed in polypropylene cages (two per cage) under consistent conditions, including a 12-hour light/dark cycle and under a controlled temperature of 25±2°C. All procedures were conducted in accordance with CPCSEA guidelines and approved by the IAEC (Approval No. IAEC/CCP/Feb 2023/13). Obesity was induced using a high-fat diet (HFD), formulated with slight modifications from the composition reported by Srinivasan et al¹⁸.

Experimental design and treatment:

This study involved nine groups of six rats each to assess the effects against obesity for the plant Ficus benjamina in High Fat Diet induced obesity. Group I served as the normal control, while Group II received only an HFD. Group III was treated by using orlistat (30 mg/kg, i.p.) alongside HFD. Groups IV–IX received hydroethanolic extract (F1) and ethyl acetate fraction (F3) of F. benjamina at 100, 200, and 400mg/kg doses orally with HFD¹⁹. Anthropometric Parameters including BMI, body weight, and Lee index were measured at weeks 4 and 8. Blood samples were collected for biochemical analysis, and adipose tissues were excised, weighed, and examined histologically. Remaining tissues were analyzed for antioxidant (TBARS, SOD, GSH) and liver enzyme (AST, ALT, ALP) levels using standard assay kits²⁰.

IN-SILICO STUDIES: Ligand and Protein Preparation:

Molecular docking was conducted to assess interactions between Ficus benjamina phytoconstituents and selected target receptors. Ligands were prepared using ChemDraw and converted to SDF format. Docking was performed using AutoDock Vina via PyRx 0.8, with additional analysis through ChemOffice 2016 and Discovery Studio 2020^21-23. Simulations were executed on a DELL workstation with Intel Core i5-12400 CPU ,Ubuntu 22.04 LTS, 8 GB RAM, and an 4 GB Nvidia GeForce RTX 3050 GPU²⁴. The 3D structure of pancreatic lipase (1LPB) was retrieved from Protein Data Bank and then prepared using the Discovery Studio by removing ligands, water, and heteroatoms, followed by hydrogen addition and charge optimization²⁵. Docking was performed in PyRx using AutoDock Vina, with orlistat as the reference. Phytochemicals from Ficus benjamina were converted to PDBQT format and evaluated for binding affinity to assess their pharmacological potential.

RESULTS:

Phyto-chemical evaluation and HPTLC profiling of Leaves extract of F. benjamina:

Phytochemical evaluation of Ficus benjamina leaf extract (F1) confirmed the occurrence of tannins, alkaloids, flavonoids, saponins, and phenolics. HPTLC analysis identified six components, with dominant peaks at Rf values 0.30 and 0.73, corresponding to rutin and quercetin, showing peak areas of 52.22% at 254 nm and 59.28% at 366nm, respectively.

Figure 1-2: Chromatogram of the standards Rutin and Quercetin

Effect of Ficus benjamina extracts on pancreatic lipase inhibition assay:

The anti-obesity potential of Ficus benjamina hydroalcoholic extract and its fractions was evaluated via pancreatic lipase inhibition. Both, crude extract and the ethyl acetate fraction showed dose-dependent inhibition. IC₅₀ values were 74.81µg/mL for extract and 65.48µg/mL for ethyl acetate fraction, compared to 19.13µg/mL for orlistat.

Figure 3. Pancreatic lipase enzyme Inhibition by extract and its different fractions. Orlistat was utilized as positive control *p<0.05.

In-vivo Anti-Obesity Activities:

Change in Body weight, Body mass index and Lee index:

Compared to the normal control group, HFD-fed rats showed a statistically significant (p<0.05) rise in body weight, Lee index, and body mass index. Orlistat effectively reduced these parameters. Similarly, treatment with Ficus benjamina fractions F1 (Hydroalcoholic extract of leaves of F. benjamina) and F3 (Ethyl fraction of leaves of Ficus benjamina.) (100, 200, and 400 mg/kg) induced dose-dependent reduction (p<0.05) in body weight, Lee index, and BMI, comparable to orlistat. (Table 1)

Table 1. Effect of F1 F3 on body weight, BMI and Lee Index

Variables	Initial Body weight(g)	Final Body weight(g)	BMI (g/cm²)	*Lee Index(g^1/3/cm1000)**
NC	227.5 ± 3.1	281.8 ± 2.7	1.35 ± 0.06	281.82 ± 4.7
HFD	228.3 ± 4.3	405.7 ± 9.3^a	1.91 ± 0.08^a	360.50 ± 12.3^a
HFD+ Orlistat	229.1 ± 3.8	289.5 ± 7.6^b	1.34 ± 0.06^b	285.20 ± 8.5^b
F1+Orlistat(Low)	228.3 ± 4.2	393.5 ± 15.9	1.82 ± 0.09	352.40 ± 12.2
F1+Orlistat(Medium)	231.6 ± 3.4	340.8 ± 9.19^b	1.67 ± 0.08^b	329.20 ± 8.79^b
F1+Orlistat(High)	233.3 ± 5.2	314.5 ± 8.95^b	1.50 ± 0.09^b	301.30 ± 7.19^b
F3+Orlistat(Low)	228.3 ± 4.6	388.3 ± 5.7	1.88 ± 0.10	352.1 ± 11.6
F3+Orlistat(Medium)	235.8 ± 6.6	322.8 ± 10.2^b	1.65 ± 0.09^b	324.30 ± 9.8^b
F3+Orlistat(High)	232.5 ± 3.9	302.8 ± 12.3^b	1.46 ± 0.08^b	293.60 ± 10.7^b

Findings are expressed as Mean±SD;^a=p<0.01 vs NC; ^b=p<0.05 vs HFD control: Normal Control; NC, High Fat Diet; HFD

Table 2: Effect of F1 and F3 on the serum glucose and serum lipid profile

Variables	NC	HFD	HFD+Orlistat	HFD +F1 (Low)	HFD +F1 (Medium)	HFD +F1 (High)	HFD +F3 (Low)	HFD +F3 (Medium)	HFD +F3 (High)
GLU (mg/dl)	89.17 ± 3.08	170.38 ± 11.4^a	98.5 ± 3.6^b	166.2 ± 10.3	137.8 ± 5.2^b	105.2 ± 3.6^b	162.3 ± 7.8	135.3 ± 6.5^b	102.20 ± 3.8^b
TC (mg/dl)	89.17 ± 4.03	170.30 ± 13.26^a	98.50 ± 5.03^b	166.20 ± 8.21	137.86 ± 5.25^b	105.20 ± 6.10^b	162.30 ± 9.67	135.30 ± 6.72^b	102.20 ± 5.8^b
TG (mg/dl)	69.71 ± 2.90	165.33 ± 15.21^a	78.50 ± 4.89^b	126.83 ± 9.23	112.30 ± 5.50^b	99.30 ± 5.12^b	122.67 ± 5.02	104.50 ± 4.54^b	94.50 ± 5.95^b
HDL (mg/dl)	47.33 ± 3.76	15.20 ± 1.24^a	43.40 ± 3.29^b	20.50 ± 2.12	28.89 ± 1.25^b	36.83 ± 2.76^b	22.50 ± 1.16	29.67 ± 2.07^b	38.33 ± 3.14^b
VLDL (mg/dl)	13.83 ± 1.01	33.07 ± 1.9^a	15.70 ± 0.90^b	25.37 ± 1.10	22.46 ± 1.03^b	19.86 ± 0.87^b	24.53 ± 1.32	20.90 ± 0.93^b	18.90 ± 1.08^b
LDL (mg/dl)	28.01 ± 2.14	122.03 ± 8.08^a	39.40 ± 3.12^b	120.83 ± 5.22	86.45 ± 5.76^b	48.51 ± 3.05^b	115.27 ± 6.98	84.73 ± 4.11^b	44.97 ± 8.28^b

Findings are expressed as Mean±SD;^a=p<0.01 vs NC; ^b=p<0.05 vs HFD control: Normal Contol; NC, High Fat Diet; HFD, F1 Hydroalcoholic extract of leaves of Ficus benjamina, F3 Ethyl fraction of leaves of the Ficus benjamina; GLU; Serum glucose, TC; Total Cholesterol, TG; Triglyceride, HDL; High density lipoproteins, VLDL; Very low density lipoproteins, LDL; Low density lipoproteins.

Table 3: Effect of F1 and F3 on different fat depot and serum liver enzymes

Parameters	MES	RET	EPI	TF	AST	ALT	ALP
NC	2.91 ± 0.05	2.96 ± 0.08	2.64 ± 0.06	8.51 ± 1.3	28.98 ± 1.65	21.67 ± 1.67	70.23 ± 4.38
HFD	9.30 ± 0.96^a	11.45 ± 1.26^a	16.21 ± 1.1	36.96 ± 3.8^a	70.56 ± 5.08	67.50 ± 3.91^a	142.76 ± 9.29^a
HFD+Orlistat	3.70 ± 0.7^b	4.20 ± 0.6^b	2.98 ± 0.5	10.88 ± 1.2^b	32.56 ± 2.2	27.80 ± 1.9^b	77.67 ± 3.8^b
HFD +F1(Low)	9.0 ± 0.9	10.90 ± 1.21	15.27 ± 2.35	35.17 ± 5.3	68.90 ± 3.5	63.20 ± 5.4	136.54 ± 9.32
HFD +F1 (Medium)	7.10 ± 0.8^b	8.86 ± 0.5^b	11.13 ± 1.04	27.08 ± 4.3^b	52.70 ± 3.87	49.67 ± 3.3^b	108.40 ± 5.3^b
HFD +F1 (High)	5.10 ± 0.4^b	6.20 ± 0.6^b	8.56 ± 1.12	19.86 ± 3.2^b	39.50 ± 3.12	32.65 ± 2.4^b	91.43 ± 6.8^b
HFD +F3(Low)	8.90 ± 0.9	10.46 ± 1.05	14.78 ± 1.7	34.14 ± 4.4	66.89 ± 4.4	60.76 ± 3.1	132.78 ± 8.01
HFD +F3 (Medium)	6.90 ± 0.8^b	8.56 ± 0.9^b	10.77 ± 0.8	26.23 ± 3.9^b	50.77 ± 2.6	43.87 ± 2.9^b	100.45 ± 4.1^b
HFD +F3 (High)	4.60 ± 0.5^b	5.67 ± 0.3^b	6.43 ± 0.7	16.70 ± 2.5^b	36.70 ± 1.9	28.40 ± 2.1^b	85.98 ± 6.6^b

Findings are expressed as Mean±SD;^a=p<0.01 vs NC; ^b=p<0.05 vs HFD control. NC: Normal Contol: HFD; High Fat Diet: F1: Hydroalcoholic extract of leaves of Ficus benjamina; F3: Ethyl fraction of leaves of Ficus benjamina; Mesenteric: MES; Retroperitoneal: RET; Epididymal: EPI; Total Fat: TF

Changes in the serum biochemical markers:

A High-Fat Diet (HFD) significantly (p<0.05) elevated various markers like TC, TG (etc) levels while reducing the HDL levels compared to normal controls. Orlistat, used as a reference, effectively normalized these parameters. Treatment with Ficus benjamina extracts (F1 and F3) at 100, 200, and 400 mg/kg/day has resulted dose-dependent improvement in lipid profile and glucose levels, showing comparable efficacy to orlistat. (Table 2)

Changes observed in the weight of different fat depots:

Rats fed with High Fat Diet, has shown a significant increase in retroperitoneal, epididymal, and mesenteric fat pad weights. Treatment with Ficus benjamina fractions F1 and F3 (100, 200, and 400 mg/kg/day) significantly (p<0.05) reduced fat pad weights with increasing dose as compared to the HFD control group (Table 3)

Effect on plasma liver function:

HFD-fed rats exhibited a significant (p<0.05) increase in the liver enzymes: AST, ALT, and ALP in contrast to the normal control group. Orlistat significantly improved these markers. Similarly, treatment with Ficus benjamina fractions F1 and F3 (100, 200, and 400 mg/kg/day) led to a dose-dependent, significant (p<0.05) reduction of liver enzyme levels, indicating improved hepatic function (Table 3).

HISTOPATHOLOGY:

Adipose tissue in the normal group showed well-organized architecture with normal adipocytes and minimal fat accumulation. HFD-fed rats displayed significant adipocyte hypertrophy and fat deposition. Treatment with orlistat reduced adipocyte size and fat accumulation. Similarly, Ficus benjamina extracts F1 and F3 (100, 200, 400 mg/kg/day) significantly (p<0.05) improved adipose morphology and reduced fat deposition, comparable to the standard drug.

Figure 4. Histopathology studies.

A- histopathological sample of rat tissue of normal group which shows normal cell structure. B- histological sample of rat tissue, treated with high fat diet (HFD) which shows significant cellular degeneration and adipose tissue deposition. C- histological sample of rat tissue treated with standard drug orlistat (30mg/kg), which shows decreased of cellular degeneration and adipose tissue deposition. D- show Low dose (100mg/kg)E- shows treatment with medium dose (200mg/kg) and F- shows treatment with high dose (400mg/kg).

COMPUTATIONAL STUDIES:

Docking scores, including hydrogen and hydrophobic interactions with corresponding bond lengths between target proteins and selected phytoconstituents, is -5.7,-7.7,-9.4 and -8.7kcal mol^-1 for caffeic acid, quercetin, ruin and stigmasterol respectively. Complex formation scores were calculated using AutoDock Vina in PyRx and visualized with Discovery Studio.

DISCUSSION:

This study explored the anti-obesity potential of Ficus benjamina leaf extract using in vitro, in vivo, and in silico approaches. The hydroalcoholic extract (F1) and its ethyl acetate fraction (F3) were assessed for pancreatic lipase inhibition and their impact on lipid metabolism in HFD-induced obese rats. The phytochemical evaluation confirmed alkaloids, flavonoids, phenolics, tannins, and saponins in the crude extract, with HPTLC identifying rutin and quercetin as major constituents^26,27.

Both F1 and F3 showed dose dependent lipase inhibitory action, with IC₅₀ values of 74.81 and 65.48µg/mL, respectively, compared to 19.13µg/mL for orlistat. F1 and F3 significantly (p<0.05) reduced the Body weight, Body Mass Index, Lee index, and fat depot weights in increasing dose-dependent manner, comparable to orlistat. They also improved lipid profiles and normalized liver enzymes (AST, ALT, ALP), indicating hepatoprotective effects. Histological analysis confirmed reduced adipocyte size and fat accumulation in treated groups.

Molecular docking revealed strong binding affinities of quercetin and rutin to obesity-related targets, supporting their role in the observed pharmacological activity. These findings highlight Ficus benjamina as a promising candidate for managing obesity and related metabolic disorders.

CONCLUSION:

The hydroethanolic extract of Ficus benjamina (HELEFB) demonstrated promising anti-obesity potential in HFD -induced obese rats. The extract significantly improved overall metabolic health, including weight management, lipid regulation, and antioxidant defence. Its therapeutic effects on reducing oxidative stress and enhancing liver function further highlight its potential as a natural remedy for combating obesity and associated metabolic disorders. These findings suggest that Ficus benjamina could be an effective, plant-based intervention for obesity management.

ACKNOWLEDGMENT:

Author would like to acknowledge Chandigarh University, Gharuan and Chandigarh College of Pharmacy, Landran, Mohali.

CONFLICT OF INTEREST:

Author declares no conflicts of interest.

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Received on 06.04.2025 Revised on 09.08.2025

Accepted on 15.11.2025 Published on 03.04.2026

Available online from April 06, 2026

Research J. Pharmacy and Technology. 2026;19(4):1625-1630.

DOI: 10.52711/0974-360X.2026.00232

This work is licensed under a Creative Commons Attribution-NonCommercial-ShareAlike 4.0 International License. Creative Commons License.