Effect of Qizhu prescription on a mouse model of non-alcoholic fatty liver disease incluced by high-fat,high-fructose,and high-cholesterol diet and its mechanism
- VernacularTitle:芪术方对高脂高果糖高胆固醇诱导的非酒精性脂肪性肝病小鼠模型的影响及其机制
- Author:
Jiahao CHEN
1
;
Zhenhua ZHOU
Author Information
- Keywords: Non-alcoholic Fatty Liver Disease; Qi Zhu Formula; Mitophagy; Lipid Metabolism; Inflammation
- From: Journal of Clinical Hepatology 2024;40(11):2205-2212
- CountryChina
- Language:Chinese
- Abstract: Objective To investigate the therapeutic effect and mechanism of action of Qizhu prescription in mice with non-alcoholic fatty liver disease(NAFLD).Methods A total of 60 male C57BL/6J mice were randomly divided into normal group,model group,low-dose Qizhu prescription group(4.75 g/kg),middle-dose Qizhu prescription group(9.50 g/kg),high-dose Qizhu prescription group(19.00 g/kg),Yishanfu group(228 mg/kg),with 10 mice in each group.After 16 weeks of modeling with a high-fat high-cholesterol diet and 20%fructose water,each group was given the corresponding drug once a day for 8 weeks.The serum levels of alanine aminotransferase(ALT),aspartate aminotransferase(AST),total cholesterol(TC),triglyceride(TG),and low-density lipoprotein(LDL)were measured;ELISA was used to measure the serum levels of free fatty acid(FFA),tumor necrosis factor-α(TNF-α),interleukin-1β(IL-1β),superoxide dismutase(SOD),and malondialdehyde(MDA);HE staining and oil red O staining were used to the pathological changes of liver tissue;Western blot was used to measure the protein expression levels of LC3BⅡ/Ⅰ,p62/SQSTM1,Beclin-1,and Drp1,and real-time PCR was used to measure the mRNA expression levels of Drp1,Beclin-1,and p62/SQSTM1.A one-way analysis of variance was used for comparison of continuous data between multiple groups,and the least significant difference t-test was used for further comparison between two groups.Results Compared with the normal group,the model groups had significant increases in the serum levels of TG,TC,ALT,AST,LDL,FFA,TNF-α,IL-1β,and MDA and a significant reduction in the serum level of SOD(P<0.05).HE staining showed that the mice in the model group had hepatocyte steatosis and a large number of fat vacuoles in liver tissue,and oil red O staining showed that the mice in the model group had a large number of red lipid droplets of varying sizes in hepatocytes,with a significant increase in the percentage of oil red O staining area compared with the normal group(P<0.05).Real-time PCR showed that compared with the normal group,the model group had significant increases in the mRNA expression levels of Drp1 and Beclin-1 and a significant reduction in the mRNA expression level of p62/SQSTM1 in liver tissue(all P<0.05),and Western blot showed that compared with the normal group,the model group had significant increases in the protein expression levels of Drp1,Beclin-1,and LC3BⅡ/Ⅰ and a significant reduction in the protein expression level of p62/SQSTM1 in liver tissue(all P<0.05).Compared with the model group,some Qizhu prescription groups and the Yishanfu group had significant reductions in the serum levels of TG,TC,ALT,AST,LDL,FFA,TNF-α,IL-1β,and MDA and a significant increase in the serum level of SOD(all P<0.05).Compared with the model group,each administration group had a significant improvement in steatosis of liver tissue,a significant reduction in the percentage of oil red O staining area,significant reductions in the mRNA expression levels of Drp1 and Beclin-1,and a significant increase in the mRNA expression level of p62/SQSTM1(all P<0.05);there were significant reductions in the protein expression levels of Drp1,Beclin-1,and LC3BⅡ/Ⅰ,while some administration groups had a significant increase in the protein expression level of p62/SQSTM1(all P<0.05),with a significantly better effect in the middle-and high-dose Qizhu prescription groups(all P<0.01).Conclusion Qizhu prescription improves lipid metabolism and inflammation in mice with NAFLD possibly by regulating hepatocyte mitophagy.
