Sida rhomboidea. Roxb leaf extract down-regulates expression of PPARγ2 and leptin genes in high fat diet fed C57BL/6J Mice and retards in vitro 3T3L1 pre-adipocyte differentiation

Abstract

Sida rhomboidea. Roxb leaf extract (SRLE) is being used by the populace of North-East India to alleviate symptoms of diabetes and obesity. We have previously reported its hypolipidemic and anti-diabetic properties. In this study, we report the effect of SRLE on (i) in vivo modulation of genes controlling high fat diet (HFD) induced obesity and (ii) in vitro 3T3L1 pre-adipocyte differentiation and leptin release. Supplementation with SRLE significantly prevented HFD induced increment in bodyweight, plasma lipids and leptin, visceral adiposity and adipocyte hypertrophy. Also, SRLE supplementation reduced food intake, down regulated PPARγ2, SREBP1c, FAS and LEP expressions and up-regulated CPT-1 in epididymal adipose tissue compared to obese mice. In vitro adipogenesis of 3T3L1 pre-adipocytes was significantly retarded in the presence of SRLE extract. Also decreased triglyceride accumulation, leptin release and glyceraldehyde-3-Phosphate dehydrogenase activity along with higher glycerol release without significant alteration of viability of 3T3L1 pre-adipocytes, was recorded. Our findings suggest that prevention of HFD induced visceral adiposity is primarily by down regulation of PPARγ2 and leptin gene expression coupled with attenuation of food intake in C57BL/6J mice. SRLE induced prevention of pre-adipocytes differentiation, and leptin release further substantiated these findings and scientifically validates the potential application of SRLE as a therapeutic agent against obesity.

Keywords: 3T3L1 cells; PPARγ2; Sida rhomboidea. Roxb; leptin; obesity.

Figure 1

Effect of S. rhomboidea. Roxb leaf extract feeding on (A) body weight gain and (B) food intake. Results are expressed as mean ± S.E.M., n = 6. Where, time points not sharing common letter indicate significant differences (p < 0.05).

Figure 2

Effect of S. rhomboidea. Roxb leaf extract feeding on morphological and anatomic evaluation of visceral adiposity in Lean (A and D), OB (B and E) and OB + SRLE (C and F) groups and abdominal, epididymal and perirenal fat pad weights. Results are expressed as mean ± S.E.M., n = 6. Where, timepoints not sharing common letter indicate significant differences (p < 0.05).

Figure 3

Effect of S.rhomboidea.Roxb leaf extract feeding on quantitative RT-PCR analysis of PPARγ2, SREBP1c, CPT-1 FAS and LEP mRNA expression. Results are expressed as mean ± S.E.M., n = 6. Where, timepoints not sharing common letter indicate significant differences (p < 0.05).

Figure 4

Effect of S.rhomboidea.Roxb leaf extract on cell viability. Results are expressed as mean ± S.E.M., n = 3. Where, * P < 0.05, ** P < 0.01 and P < 0.001 and ^ns non significant compared to 0 μg/mL SRLE.

Figure 5

Photomicrograph of Oil Red O stained differentiating 3T3L1 cells (A) untreated, (B) treated with 10 μg/mL SRLE, (C) treated with 20 μg/mL SRLE, (D) treated with 50 μg/mL SRLE, (E) treated with 100 μg/mL SRLE and (F) treated with 200 μg/mL SRLE and qualitative (whole well image of Oil Red O stained adipocytes) and quantitative (% adipogenesis) evaluation of adipocyte differentiation (G and H). Results are expressed as mean ± S.E.M., n = 3. Where, * P < 0.05, ** P < 0.01 and P < 0.001 and ^ns non significant compared to 0 μg/mL SRLE.

Figure 6

Effect of S. rhomboidea. Roxb leaf extract on in vitro (A) leptin release, (B) triglyceride accumulation, (C) glycerol release, and (D) G3PDH activity during 3T3L1 pre-adipocyte differentiation were evaluated as described in materials and methods. Results are expressed as mean ± S.E.M., n = 3. Where, * P < 0.05, ** P < 0.01 and P < 0.001 and ^ns non significant compared to 0 μg/mL SRLE.