Effect of Trifolium pratense extract on methionine-choline-deficient diet-induced steatohepatitis in C57BL/6 mice.
OBJECTIVE
The potential of Trifolium pratense (red clover) extract in the prevention of lipid disorder has attracted increasing attention in recent years. In this study, the aim was to determine whether and how red clover extract affected the development of murine diet-induced non-alcoholic steatohepatitis.


METHODS
Non-alcoholic steatohepatitis was induced in C57BL/6 mice by feeding mice with a methionine-choline-deficient (MCD) diet. Hematoxylin and eosin staining was used for histological analyses. Real-time PCR was used to analyze the mRNA expression levels.


RESULTS
Hepatic steatosis and necroinflammation was observed in MCD diet-fed mice, and this diet-induced steatosis was significantly attenuated, whereas liver inflammation was not significantly attenuated, by red clover extract treatment. Consistent with the results of H&E staining, the MCD diet-induced increase of liver triglycerides and cholesterol levels were significantly reduced by red clover extract treatment. However, with the improvement in hepatic steatosis, mRNA levels of acetyl CoA oxidase, carnitine palmitoyl transferase-1, and liver fatty acid-binding protein, three genes regulated by peroxisome proliferator-activated receptor (PPAR) α, were unaffected.


CONCLUSIONS
Red clover extract alleviated MCD diet-induced hepatic steatosis, but did not ameliorate liver inflammation in C57BL/6 mice, and the improvement in hepatic steatosis was not through activating PPARα.
School of Life Sciences, Longyan University, Longyan 364000, China; Fujian Key Laboratory of Preventive Veterinary Medicine and Biotechnology, Longyan 364000, China.
Tong
T
Chen
School of Life Sciences, Longyan University, Longyan 364000, China.
Fo-Jin
FJ
Zhong
School of Life Sciences, Longyan University, Longyan 364000, China.
Ya-Min
YM
Hong
School of Life Sciences, Longyan University, Longyan 364000, China.
Wei-Jiao
WJ
Su
School of Life Sciences, Longyan University, Longyan 364000, China.
Li-Li
LL
Zhuang
School of Life Sciences, Longyan University, Longyan 364000, China; Fujian Key Laboratory of Preventive Veterinary Medicine and Biotechnology, Longyan 364000, China. Electronic address: qlongxin@tom.com.
Long-Xin
LX
Qiu
D000818:Animals; D002784:Cholesterol / Q000378:metabolism; D002796:Choline Deficiency / Q000150:complications*; D004032:Diet / Q000009:adverse effects; D004195:Disease Models, Animal; D007249:Inflammation / Q000188:drug therapy / Q000378:metabolism; D008099:Liver / Q000187:drug effects* / Q000378:metabolism; D008297:Male; D008715:Methionine / Q000172:deficiency*; D051379:Mice; D008810:Mice, Inbred C57BL; D065626:Non-alcoholic Fatty Liver Disease / Q000188:drug therapy* / Q000209:etiology / Q000378:metabolism; D047495:PPAR gamma / Q000378:metabolism; D008517:Phytotherapy; D010936:Plant Extracts / Q000494:pharmacology / Q000627:therapeutic use*; D012333:RNA, Messenger / Q000378:metabolism; D029921:Trifolium; D014280:Triglycerides / Q000378:metabolism
D047495:PPAR gamma; D010936:Plant Extracts; D012333:RNA, Messenger; D014280:Triglycerides; D002784:Cholesterol; D008715:Methionine
Hepatic steatosis; Liver inflammation; Peroxisome proliferator-activated receptor α; Steatohepatitis; Trifolium pratense