Preventive Of Jiawei Sijunzi Decoction On Nonalcoholic Fatty Liver In Mice

Nonalcoholic fatty liver disease(NAFLD)is a clinicopathological syndrome characterized by excessive fat deposition in hepatocytes.Chinese herbal medicine has received increasing attention as a potential therapeutic drug for NAFLD in the past few decades.We abide by the basic theory of traditional Chinese medicine and the research results of other scholars,adding or subtracting herbs and dosage based on Sijunzi Decoction.According to the relationship between Jun,Chen,Zuo and Shi,the traditional Chinese medicine formula-Jiawei Sijunzi Decoction(CHF)was formed.The purpose of this study is to determine the preventive effect of Jiawei Sijunzi Decoction on NAFLD and explore its mechanism of action,and to provide new ideas for the subsequent development of NAFLD prevention and treatment drugs.In this study,we explore the preventive effect of CHF on NAFLD and its molecular mechanism through in vivo experiments and in vitro experiments.1.In vivo experiments: Mice NAFLD model was established by feeding high-fat diet.After 1 w of adaptive feeding,we randomly divided healthy male Kunming mice into 5 groups,in which the control group(CON)was fed with ordinary diets and the model group(HFD)was fed with high-fat diets.The CHFtreated group was fed with high-fat diets and treated with low-concentration(HFD+L),mediumconcentration(HFD+M),and high-concentration(HFD+H)CHF for 8 weeks by gavage.The body weight of each group of mice was measured weekly during the experiment.After the test,the blood and liver of the mice were collected;the relevant biochemical indicators in the blood and liver were measured,and the key proteins and genes in the ERs,NF-?B and SREBP-1c pathway were measured.2.In vitro experiments: Cell models were established by stimulating liver cells of AML 12 with FFA.We first used different concentrations of FFA and CHF to treat the cells respectively,and selected the optimal treatment concentration by MTT experiment.The experiment was divided into 5 groups according to different treatments: the control group(CON)cells were not treated;the model group(FFA)cells were induced with FFA,at the same time,the CHF treatment groups were added with low concentration(FFA+L)and medium concentration(FFA+M)and high concentration(FFA+H)CHF for 24 hs.Cells were collected after the above treatment,and the key proteins and genes in the ERs,NF-?B and SREBP-1c pathways were detected.In vivo experiment results: 1.Oral administration of CHF at different concentrations can improve pathological changes such as vacuolation,inflammatory infiltration,and lipid deposition in the liver of mice caused by high-fat diet feeding.The oral administration of high-concentration CHF significantly inhibited the liver index of mice(P <0.05).2.The oral administration of medium and high concentration CHF significantly inhibited the increase of TC content in serum(P <0.001)and liver(P <0.05)of mice fed a high-fat diet.Western blot results showed that,compared with the HFD group,oral administration of different concentrations of CHF significantly inhibited the increase in the relative expression of SREBP-1c protein in mouse liver(P<0.001);at the same time,the relative expression of ACC1 protein in the liver of mice in HFD+M(P<0.05)and HFD+H(P<0.001)groups,the relative expression of FAS protein in the liver of mice in HFD+L(P<0.05),HFD+M(P<0.001),and HFD+H(P<0.001)groups was significantly lower than that in HFD group.In addition,The changes of the relative expression levels of Fas and Acc1 genes in the liver of mice in different concentrations of CHF treatment group were basically consistent with the protein change trend.3.CHF treatment can improve the changes of SOD,CAT activity and MDA levels in the liver of mice.Western blot results showed that compared with the HFD group,the relative expression levels of PERK protein in the liver of mice in the HFD+M(P<0.01)and HFD+H(P<0.01)groups were significantly reduced;The relative expression levels of IRE1 protein in the livers of mice in HFD+L(P<0.01),HFD+M(P<0.001)and HFD+H(P<0.001)groups were significantly reduced;the relative expression of CHOP protein in livers of HFD+M and HFD+H mice was significantly reduced(P<0.01).4.The treatment of CHF can improve the changes of inflammatory factors such as TNF-?,IL-6 and the liver injury indexes such as AST and ALT in the serum and liver of mice;in addition,compared with the HFD group,the liver ALT levels in the HFD+L(P<0.01),HFD+M(P<0.05),and HFD+H(P<0.01)groups were significantly reduced.Western blot results showed that the relative expression levels of IKK-? protein in the liver of mice in HFD+L(P<0.001),HFD+M(P<0.001)and HFD+H(P<0.001)groups were significantly lower than those in HFD groups;The relative expression of NF-?B protein in liver of HFD+H group mice was significantly lower than that of HFD group(P<0.05).The PCR results showed that treatment with different concentrations of CHF can effectively improve the relative expression of I?b-? and Ikk-? genes in the liver of mice caused by high-fat diet feeding.In vitro experiment results: CHF treatment can effectively prevent FFA-induced lipid deposition around the cell;at the same time,CHF significantly inhibits the relative expression of key proteins and genes in the SREBP-1c pathway,and effectively inhibits the changes in the relative expression of PERK,IRE1,and CHOP proteins;CHF Treatment can also significantly inhibit the relative expression of NF-?B pathway-related proteins and genes in cells.The final results of in vitro experiments are basically consistent with in vivo experiments.In summary,we have studied the preventive effect and mechanism of Jiawei Sijunzi Decoction in NAFLD through in vivo and in vitro experiments.The results show that the Jiawei Sijunzi Decoction has a certain preventive on NAFLD,and this effect may be achieved by regulating key proteins and genes in the ERs,NF-?B and SREBP-1c pathways.