A herb mixture to ameliorate non-alcoholic fatty liver in rats fed a high-fat diet

Abstract

This study was performed to investigate the effects of an herb extract mixture (HM) in ameliorating non-alcoholic fatty liver disease (NAFLD). The HM contained equal amounts of 70% ethanol extracts from Zingiber officinale, Centella asiatica, and Boehmeria nivea. In vitro, the HM significantly inhibited lipid accumulation in oleic acid-stimulated HepG2 cells. We further evaluated the anti-NAFLD activities of the HM in vivo in an animal model. Rats were fed two different amounts of the HM (50 and 200 mg/kg body weight) along with a high-fat diet for 6 weeks. HM supplementation reduced liver weight; epididymal, peri-renal, and intra-abdominal fat content; and serum triglyceride, total cholesterol, and low-density lipoprotein cholesterol levels as well as increased high-density lipoprotein cholesterol levels in a dose-dependent manner. Histological evaluation of liver specimens further demonstrated that administration of HM significantly prevented hepatic lipid accumulation and subsequent development of hepatic steatosis. These findings suggest that HM can be used as an alternative nutraceutical for ameliorating NAFLD.

Keywords: Boehmeria nivea; Centella asiatica; Combinatorial herb extract; Non-alcoholic fatty liver disease; Zingiber officinale.

Fig. 1

Effect of the herb extract mixture (HM) on lipid accumulation in HepG2 cells. The cells were treated with individual herbs (Zingiber officinale [Z], Centella asiatica [C], Boehmeria nivea [B]; 50 and 100 μg/ml) or their combination (HM; 50 and 100 μg/ml) in the presence of oleic acid (OA; 0.6 mM) for 24 h, and stained with Oil Red O. (A) Lipid droplets observed by microscopy. (B) Quantitative assessment of the percentage of lipid accumulation based on Oil Red O staining. Data are expressed as the mean ± SD. ^###p < 0.001 compared to the OA group.

Fig. 2

Effect of combinatorial herb mixture (HM) on the weights of the liver, epididymal fat, peri-renal fat, and intra-abdominal fat tissues in rats. (A) Body weight and (B) feed intake. (C) Liver weight (D) epididymal fat weight (E) peri-renal fat weight and (F) intra-abdominal fat weight. Data are expressed as the mean ± SD (n = 8). ^##p < 0.01; ^###p < 0.001 compared to the high-fat diet (HFD) group.

Fig. 3

Effect of a combinatorial herb mixture (HM) on (A) serum triglyceride (B) total cholesterol (C) high-density lipoprotein (HDL)-cholesterol and (D) low-density lipoprotein (LDL)-cholesterol levels in rats. Data are expressed as mean ± SD (n = 8). ^#p < 0.05; ^###p < 0.001 compared to the high-fat diet (HFD) group.

Fig. 4

Effect of a combinatorial herb extracts mixture (HM) on histological changes in high-fat diet (HFD)-induced non-alcoholic fatty liver disease (NAFLD). (A) Liver and (B) epididymal white adipose tissue (eWAT) stained with hematoxylin and eosin and observed microscopically at 100 × and 400 × magnification. (C) Adipocyte area in the eWAT measured using ImageJ software. ND: normal diet; HFD: high-fat diet; HM 50: HFD +50 mg/kg herb mixture; HM 200: HFD +200 mg/kg herb mixture. Data are expressed as the mean ± SD (n = 8). ^###p < 0.001 compared with the HFD group.

Fig. 5

The high-performance liquid chromatography (HPLC) chromatogram of standard solution and three-herb mixture extracts. (A) HPLC chromatograms of standard solution. (B) HPLC chromatograms of three-herb mixture extracts.