Mechanism of Jiangan Jiangzhi Pill NRF2 pathway in the treatment of steatohepatitis in mice

Objective observe the healthy liver cholesterol-lowering pills for methionine choline lack of feed (Methionine-choline deficient，MCD) diet induced nonalcoholic fatty hepatitis (Nonalcoholic steatohepatitis，NASH) mechanism of lipid metabolism and inflammation in mice. Methods The mice were randomly divided into control group, model group, positive drug group, low, medium and high dose groups of Jianganjiangzhi pills, 10 mice in each group.The control group ate normal diet, and the other experimental groups ate MCD diet.The positive drug group was intritoneally injected with 150mg/kg polyene phosphatidylcholine, the low, medium and high dose groups of Jiangan Jiangzhi pills were administrated with 100 mg/kg, 200 mg/kg and 400 mg/kg respectively, once a day, and the control group and model group were given equal volume of normal saline in parallel.Weigh and record every 7 days.It lasts six weeks.The histopathological changes of liver were observed by HE staining.Serum alanine aminotransferase (ALT) and aspartate aminotransferase (AST) were detected by biochemical kit,The levels of total cholesterol (TC), triglyceride (TG), superoxide dismutase (SOD), malondialdehyde (MDA) and glutathione peroxidase (GSH-Px) in liver homogenate were detected by BCA kit.The mRNA expression levels ofNrf2, Ho1, Gclc and Nqo1 in mouse liver tissues were detected by PCR,The expression level ofNRF2 pathway related protein in mouse liver was detected by Western blot . Results Compared with the normal group, the model group had serious liver pathological injury, severe destruction of liver cell structure, widening of cell interval, collapse and infiltration of a large number of inflammatory cells.In the model group, ALT, AST, TG and TC (P<0.01) levels were significantly increased, SOD activity was decreased, MDA level was increased, and GSH-Px activity was decreased. PCR results showed that mRNA expression levels ofNrf2, Ho1, Gclc and Nqo1 in mouse liver tissues were decreased.Westernblot results showed that the relative expression levels ofNRF2, HO-1,GCLC and NQO1 proteins in mice were significantly decreased. Compared with the model group, the liver cell structure of mice in positive drug group (polyene phosphatidylcholine) and Jianggan Jiangzhiwan low-dose, medium-dose and high-dose groups was more clear, with a shorter cell interval and a small amount of inflammatory cell infiltration,ALT, AST,TG and TC levels decreased significantly, SOD activity increased, MDA level decreased, GSH-Px activity increased. The mRNA expression levels ofNrf2, Ho1, Gclc and Nqo1 in liver tissues were increased, and the protein expression levels of NRF2 pathway related proteins were significantly increased. Conclusion Jiangan Jiangzhi pills(健肝降脂丸) can regulate NRF2 pathway to enhance antioxidant capacity and reduce liver steatosis and inflammation in mice with NASH