Changes in body weight and leptin: intervention of gambir extract in obese model rats

Background: The increasing prevalence of obesity is a serious health problem that requires alternative therapy. Gambir contains catechins with anti-obesity properties that can be used as an alternative therapy. Objectives: This study aims to analyze the eﬀect of gambier extract intervention on body weight and leptin in obese rat models. Methods: This research was laboratory research with a pre-post-test design. The research subjects were male Wistar rats with a total of 36 rats. Rats were divided into 6 groups, namely KN (healthy rats), K-(obese rats), KP (obese rats given Orlistat 10.8 mg/200gBB/day), P1 (obese rats given gambir extract 20 mg/200gBW/day


INTRODUCTION
Conclusions: Gambir extract intervention at 80mg/200gBW/day in obese rats can decrease weight gain and reduce leptin levels.

KEYWORDS: body weight; catechin; gambir; leptin; obesity
Article info: Article submitted on June11, 2023 Articles revised on July 11, 2023 Articles received on August 19, 2023 Obesity is a metabolic disorder characterized by increased adipose tissue mass due to a positive energy balance (1).
Obesity is a serious public health problem 2 with a Body Mass Index >30 kg/m (2).
According to the World Obesity Federation, as much as 38% of the world's population over five suffers from obesity and is expected to reach 51% in 2035.Obesity is estimated to affect nearly half of adults (49%) in America, 39% in Europe, and 11 % in Southeast Asia in 2035 (3).The prevalence of obesity in Indonesia in the age group over 18 years has increased by 7% from 2013 to 2018 (4).In 2025, 7.5 out of 10 Indonesians are expected to be obese, or 10% of male and 14,8% female of the population (5).
Obesity is a major public health problem in developed and developing countries (6).Individuals with overweight and obese nutritional status have death rates of 47.48 and 66.67 per 1000 people per year respectively (7).Recent studies found that overweight obesity contributed to 5.5% of total deaths (8).Obesity also contributes to cardiovascular disease, metabolic syndrome, type 2 diabetes mellitus, cancer, and various reproductive disorders (6).
Obesity is caused by an imbalance in incoming and outgoing energy (9).Energy i m b a l a n c e w i l l d i s r u p t t h e e n e r g y homeostasis system (10).In the case of obesity, the energy homeostasis system is disrupted, which creates excess body weight (11).The brain regulates Body weight in response to external signals to control food intake (12).In the long term, this mechanism leads to the development of obesity (13).
Research shows a close relationship between leptin levels, adiposity, and body fat percentage (14).Leptin levels indicate how much energy is stored in fat and calorie intake (15).Excessive leptin will lose normal functions such as appetite suppression and weight loss.Leptin incompetence will lead to a lack of clinical use of leptin resulting in leptin resistance which is the beginning of the development of obesity (16).
Obesity can be prevented or controlled in various ways, such as food, exercise, and medication.However, anti-obesity drugs such as Orlistat and Sibutramine have been associated with various negative side effects, including high blood pressure, constipation, dry mouth, headaches, heart attacks, and t r o u b l e s l e e p i n g ( 6 ) .I n a d d i t i o n , pharmacological therapy in the long term requires a lot of money, so an alternative pharmacological therapy that is safe, effective, and cheaper is needed, namely the Changes in body weight and leptin: intervention of gambir extract in obese model rats 133 use of phytochemical compounds (17).
Gambir is one of the typical Indonesian plants that the community has widely used as a spice and medicine (18).Gambir extract is produced from the plant's dried leaves and young twigs and is used as an astringent in Asia (19).Gambir is beneficial in various health problems, such as anti-hyperlipidemia, antiaging, reducing plaque halitosis, and oxidative stress (20).Besides that, it can also be used as a mixture of medicinal and food ingredients (21).
Gambir contains 97% catechins (23).Recent studies have shown that catechin bioactive compounds have antioxidant and anti-obesity properties (24,25).Intervention studies conducted in the obese group showed that the c o n t e n t o f c a t e c h i n s i n b e v e r a g e s significantly reduced individual weight in the intervention group by 8-10% (26).In addition, catechins can reduce leptin levels and directly reduce body weight through increased energy expenditure and fat oxidation (27).
Research on gambir extract as a treatment for obesity in reducing body weight and improving leptin has never been done in vivo.This study aims to determine the effect of gambir extract intervention on body weight and leptin in obese rat models.

RESULTS AND DISCUSSION
Obesity induction was carried out after adaptation for 7 days.The results showed that all rats fed the HFHFr diet for 4 weeks were obese.An indicator of successful obesity in rats is when 3 the Lee index is >300 g/cm .(28).The P2 group had a higher Lee index because the initial body weight was already high.Lee's index was calculated by measuring body weight and naso-anal length.This study's results align with previous studies which explained that a high-fat diet intervention succeeded in inducing obesity in rats as measured by the Lee index (29).
Giving a high-fat diet to rats will increase body weight and produce a significant increase in adiposity compared to the normal diet group (30).The underlying mechanism is a positive energy balance that results in overweight and obesity.A high-fat diet increases hypo-thalamic gene expression in increasing calorie intake so that adiposity occurs as a physiological response of the body (31).

Body Weight
The body weight of the rats was weighed before and after being given the intervention.
Gambir extract intervention in the treatment group was given after obesity was achieved.
Based on Table 2 above, it is known that there is a significant difference in the average body weight between groups of rats after the gambir extract intervention with a significance value of 0.000.The P3 and K+ groups had the lowest average body weight compared to the obesityinduced treatment group, where there was a decreased weight gain.This study is in line with the previous study which stated that giving EGCG inhibited weight gain (32).
Similar studies also show that the catechins in green tea prevented weight gain in rats on a high-fat diet (3335).
The treatment group with a dose of 80 m g / 2 0 0 g B W / d a y ( P 3 ) i n t h i s s t u d y experienced the greatest reduction in body weight gain compared to the other two doses.
The results of this study are in line with previous research which conducted two repeated studies where the first study showed results that catechins did not reduce body weight.On the contrary, after increasing the dose in the second repeat study, the results showed significant weight loss (36) and fatty acid synthesis and increases catabolism, leading to weight loss (40).

Leptin
Leptin is a hormone whose level is directly proportional to adiposity.Leptin levels were measured after 4 weeks of intervention.
Table 3 shows that all treatment groups intervening with gambir extract had lower leptin levels than the K-group.The P3 group had the lowest average leptin levels compared to the obesity-induced treatment group.Based on the One-way ANOVA test showed that there was a significant difference in the average leptin levels between groups of rats with a significance value of 0.001.This study's results align with several previous studies as shown in the following Table 4.

Result The study's distinction
The Anti-Obesity Potential of Green Tea: The Effect on Leptin and Adiponectin (27) Epigallocatechin Gallate (EGCG) rich green tea consumption reduces leptin levels and directly reduces obesity.
The intervention used is gambir which is rich in catechins.
Anti-obesity and antidiabetic effects of Yerba Mate (Ilex paraguariensis) in C57BL/6 J mice fed a high-fat diet (41) Polyphenol yerba mate herbal tea reduces leptin levels in mice fed a high -fat diet.
The intervention used is gambir, rich in catechins, and the subject is not specifically in obese rats.(27).

Green Tea
Leptin levels in the blood reflect energy stores and fat mass (44).
Obese individuals have high levels of circulating leptin which is then exacerbated by a high-fat diet (45).Leptin levels correlate with adiposity in that adipocytes secrete leptin into the bloodstream with fat mass (46).Catechins will decrease body fat mass which then triggers a decrease in adiposity (47).It will impact leptin secretion by adipose tissue, resulting in low leptin levels in the rats that were intervened with catechin-rich gambir extract.

CONCLUSION AND RECOMMENDATIONS
Gambir rich in catechins has the potential as an alternative therapy for obesity.
type of research is a laboratory experiment with a pre-post test design for calculating body weight and a post-test for checking leptin levels.This research was conducted in the Center for Food and Nutrition Studies Laboratory, Gadjah Mada University.This research was conducted in April-May 2023.The sample for this study used male Wistar rats aged 8-12 weeks with a body weight of 150-200 grams.The sample size of 36 rats was calculated using the Sample Size Calculation in Animal Studies formula.Rats were divided into 6 groups using simple random sampling.The rat group consisted of KN (healthy rats), K-(obese rats), KP (obese rats given Orlistat 10.8 mg/200gBW/day), P1 (obese rats given gambir extract 20 mg/200gBW/day), P2 (Obese rats given gambir extract 40 mg/200gBW/day), and P3 (obese rats given gambir extract 80 mg/200gBW/day).The ingredients used are G-Fit brand gambir extract obtained from CV Uncaria Herbal Indonesia, standard feed, and High Fat High Fructose (HFHFr) feed.Leptin levels were measured using the ELISA method using the Rat Leptin ELISA Kit.Adaptation was carried out for 7 days while obesity induction and intervention were carried out for 4 weeks each.Obesity modeling was done by administering HFHFr.Rats are categorized as obese if the Lee index is >300 g/cm3.Data on body weight and leptin levels were then analyzed using SPSS For Windows version 23.0 software.The different group test was conducted using One-way ANOVA and Kruskal Wallis.This study was approved by the Research Ethics Commission of the Faculty of Medicine, Universitas Sebelas Maret with number 67/UNS.27.06.11/KEP/EC/2023.
Gambir significantly decreased body weight gain and reduced leptin levels.Gambir extract intervention at a dose of 80 mg/200 gBW/day for 4 weeks had an average body weight and leptin levels lower than the K+ group and even close to the KN group.The authors suggest assessing leptin levels before and after the intervention.high fat diet.BMC Complement Altern Med.2017 Feb;17(1):122.7. Xu H, Cupples LA, Stokes A, Liu CT.Association of Obesity With Mortality Over 24 Years of Weight History: Findings From the Framingham Heart Study.JAMA Netw Open.2018 Nov 2;1(7):e184587e184587.8. Tobias DK, Hu FB.The association between BMI and mortality: implications for obesity prevention.Lancet Diabetes Endocrinol.2018 Dec 1;6(12):9167.9. Galán JPM, Ontibón-Echeverri CM, Costa MC, Batista-Duharte A, Batista VG, Mesa V, et al.Enzymatic synthesis of capric acid-rich structured lipids and their effects on mice with high-fat dieti n d u c e d o b e s i t y.F o o d R e s I n t .2021;148:110602.10.Huai P, Liu J, Ye X, Li W-Q.Association of Central Obesity With All Cause and Cause-Specific Mortality in US Adults: A P r o s p e c t i v e C o h o r t S t u d y.F r o n t C a r d i o v a s c M e d [ I n t e r n e t ] . 2 0 2 2 ; 9 ( 8 1 6 1 4 4 ) .Av a i l a b l e f r o m : https://www.frontiersin.org/articles/10.33 89/fcvm.2022.81614411.Gómez CJ, Ena J, Arévalo Lorido JC, Seguí Ripoll JM, Carrasco-Sánchez FJ, Gómez-Huelgas R, et al.Obesity is a chronic disease.Positioning statement of the Diabetes, Obesity and Nutrition Workgroup of the Spanish Society of I n t e r n a l M e d i c i n e ( S E M I ) f o r a n approach centred on individuals with o b e s i t y .R e v C l i n E s p . 2 0 2 1 Nov;221(9):50916.12. Hall KD, Farooqi IS, Friedman JM, Klein S, Loos R, Mangelsdorf DJ, et al.The energy balance model of obesity: beyond calories in, calories out.Am J Clin Nutr [Internet].2022;115(5):124354.Available from: https://academic.oup.com/ajcn/article/115/5/1243/652216 6 13.Ayogu RNB, Oshomegie H, Udenta EA.Energy intake, expenditure and balance, and factors associated with energy balance of young adults (2039 years): a r e t r o s p e c t i v e c r o s s -s e c t i o n a l community-based cohort study.BMC Nutr [Internet].2022;8(1):142.Available from: https://doi.org/10.1186/s40795-

Table 1
shows that

Table 1 . Different averages of the lee index after induction
Changes in body weight and leptin: intervention of gambir extract in obese model rats 135