祛湿活血方通过AMPK/mTOR/ULK1自噬信号通路改善代谢相关脂肪性肝病的作用机制研究OA

OBJECTIVE To investigate the mechanism of Qushi huoxue decoction in improving metabolic-associated fatty liver disease(MAFLD)based on the AMP-activated protein kinase/mammalian target of rapamycin/UNC-51-like kinase 1(AMPK/mTOR/ULK1)autophagy signaling pathway.METHODS Male C57BL/6J mice were randomly divided into normal group,model group,and low-,medium-,and high-dose Qushi huoxue decoction groups[4.10,8.20,16.40 mg/(g·d)],with 10 mice in each group.Except for the normal group,mice in the other groups were fed a high-fat diet for 12 weeks to establish the MAFLD model.Starting 24 hours after modeling,mice in each group were administered the drug solution or normal saline once daily for 6 consecutive weeks.Serum levels of total cholesterol(TC),triglycerides(TG),alanine transaminase(ALT),aspartate transferase(AST),low-density lipoprotein cholesterol(LDL-C),and high-density lipoprotein cholesterol(HDL-C)were detected according to the kit instructions.Hematoxylin-eosin(HE)staining and oil red O staining were used to observe pathological changes in mouse liver tissue.Western blot and immunofluorescence staining were used to detect the expression and positive protein expression related to the AMPK/mTOR/ULK1 autophagy signaling pathway.RESULTS Compared with the normal group,the serum levels of TC,TG,ALT,AST,and LDL-C in the model group were significantly increased,while the HDL-C level was significantly decreased(P＜0.01).HE staining showed obvious steatosis in hepatocytes of the model group,and oil red O staining showed a large amount of lipid droplet accumulation in liver tissue.Western blot and immunofluorescence staining results showed that the expression levels of p-mTOR/mTOR and p62 proteins,as well as the relative fluorescence intensities of p-mTOR and p62 proteins in liver tissue of the model group,were significantly increased/enhanced(P＜0.01),while the expression levels of p-AMPK/AMPK,ULK1,and microtubule-associated protein 1 light chain 3 Ⅱ(LC3 Ⅱ)proteins,as well as the relative fluorescence intensities of p-AMPK and LC3 proteins,were significantly decreased/weakened(P＜0.01).After intervention with Qushi huoxue decoction,the above pathological changes and abnormal indicators in liver tissue of mice in the low-,medium-,and high-dose groups were significantly reversed,with most indicators showing statistically significant differences(P＜0.05 or P＜0.01).CONCLUSIONS Qushi huoxue decoction can significantly improve liver pathology,blood lipid levels,and liver function in MAFLD model mice;this improvement may be achieved through the activation of the AMPK/mTOR/ULK1 autophagy signaling pathway.