Sipyimigwanjung-tang, a traditional herbal medication, alleviates weight gain in a high-fat diet-induced obese mice model

Abstract

Obesity leads to the development of metabolic syndrome and comorbidities. Overweight and obesity continue to be a relentless global issue. Sipyimigwanjung-tang (SGT), a traditional herbal medication, was first mentioned in Dongui Sasang Shinpyun and has been used to treat edema, meteorism, and jaundice, which are common findings associated with obesity. The main physiological feature of obesity is expanded adipose tissue, which causes several impairments in liver metabolism. Therefore, this study aimed to investigate the anti-obesity effects of SGT in the epididymal white adipose tissue (eWAT) and livers of high-fat diet (HFD)-induced obese mice. SGT significantly blocked HFD-induced weight gain in C57BL/6N mice. In addition, SGT effectively reduced the increased weight and adipocyte size in eWAT of HFD-induced obese C57BL/6 N mice. Moreover, SGT significantly decreased the elevated gene expression of Peroxisome proliferator-activated receptor γ, CCAAT/enhancer-binding protein α, and Sterol regulatory element-binding protein 1 in the eWAT of HFD-induced obese mice. Furthermore, SGT significantly decreased lipid accumulation in the livers of HFD-induced obese mice and differentiated 3T3-L1 adipocytes. Hence, the present study provides substantial evidence that SGT has potential therapeutic effects on obesity.

Keywords: Adipogenesis; High-fat diet; Obesity; Sipyimigwanjung-tang; White adipose tissue.

Graphical abstract

Fig. 1

Analysis of ingredients identified from SGT aqueous extract. HPLC chromatograms of (A) hesperidin, (B) naringin, (C) 6-gingerol, (D) honokiol, (E) magnolol, and (F) SGT.

Fig. 2

Effect of SGT on lipid accumulation in 3T3-L1 adipocytes. (A) 3T3-L1 preadipocytes were treated with various concentrations of SGT for 48 h and their viability was estimated using the MTT assay. The values are represented as mean ± S. D. *P < 0.05 and **P < 0.01 vs. untreated cells; significance was determined using one-way ANOVA followed by Dunnett's post hoc test. (B) 3T3-L1 preadipocytes were stimulated with a differentiation medium in the presence or absence of the indicated SGT concentrations for 8 days and subjected to Oil Red O staining. Stained cells were visualized under a microscope at 100× magnification. Non-adjusted images are provided as supplementary materials. (C) Oil Red O was dissolved in isopropanol and absorbance was measured at 510 nm. The values are represented as the mean ± S. D. ^##P < 0.01 vs. non-treated cells; **P < 0.01, differentiated cells; significance was determined using one-way ANOVA followed by Dunnett's post hoc test. (For interpretation of the references to colour in this figure legend, the reader is referred to the Web version of this article.)

Fig. 3

Effect of SGT on body weight gain, serum TC and TG levels in HFD-induced obese mice. (A) Mouse body weight was measured weekly for 11 weeks. (B) Body weight gain was calculated. (C) The food intake was measured weekly and calculated. (D) Serum total cholesterol and (E) triglyceride were detected. The values are represented as mean ± S. D. ^#P < 0.05, ^##P < 0.01, and ^###P < 0.001 vs. CON; *P < 0.05, **P < 0.01, and ***P < 0.001 vs. HFD group; significances were determined using two-way ANOVA followed by a Bonferroni post hoc test, and one-way ANOVA followed by a Dunnett's post hoc test.

Fig. 4

Effect of SGT on adipogenesis in eWAT of HFD-induced obese mice. At the end of the experimental period, the weights of (A) eWAT and (B) eWAT index were measured. The eWAT index is a measure of eWAT weight based on the body weight of the mice. (C) Hematoxylin/eosin staining images are shown at a magnification of 100. × (D) The average diameter of adipocytes in eWAT of each group. qRT-PCR was performed to determine mRNA levels of (E) Pparγ, (F) C/ebpα, and (G) Srebp1. The values are represented as mean ± S. D. ^#P < 0.05, ^##P < 0.01, and ^###P < 0.001 vs. CON; *P < 0.05 and **P < 0.01 vs. HFD group; significance was determined using one-way ANOVA followed by Dunnett's post hoc test.

Fig. 5

Effect of SGT on adipogenesis in liver tissue of HFD-induced obese mice. (A) At the end of the experimental period, the mouse liver was dissected for macroscopic analysis and subjected to hematoxylin/eosin staining. Images are shown at the magnification of 100×. (B) Western blot was performed to determine the protein level of PPARγ, C/EBPα, and SREBP1. The uncropped gel blots are provided as supplementary materials. qRT-PCR was performed to determine the mRNA level of (C) Pparγ, (D) C/ebpα, and (E) Srebp1. The values are represented as mean ± S. D. ^###P < 0.001 vs. control; **P < 0.01 and ***P < 0.001 vs. HFD group; significances were determined using one-way ANOVA followed by a Dunnett's post hoc test.