The Pharmacological Assessment Of Zexie-baizhu Decoction On Non-alcoholic Fatty Liver Diseases

Non-alcoholic fatty liver disease(NAFLD)is a series of progressive metabolic diseases,including Non-alcoholic Fatty Liver(NAFL),Nonalcoholic Steatohepatitis(NASH),fibrosis,cirrhosis,and hepatocellular carcinoma.Until 2019,the global prevalence of NAFLD had reached 25%,especially in China mainland,the prevalence reached up to 32.9%,which brought the whole society a heavy burden.However,the pathogenesis of NAFLD is so complicated that single-target drugs might end in unsatisfactory efficacy or even adverse effects.In this case,screening natural products with multiple ingredients and targets through effective phenotypes may be a suitable strategy for NAFLD treatment.And traditional Chinese medicine(TCM)characterized by multi-ingredients,and multi-targets has unique advantages in long-term clinical practice.Zexie-Baizhu(ZXBZ)decoction,which contains Zexie(Alisma PlantagoAquatica subsp.Orientale(Sam.)Sam.)and Baizhu(Atractylodes macrocephala Koidz.)and is prepared with water in a ratio of 5:2,is a Chinese classical formula recorded in“Synopsis of Prescriptions of the Golden Chamber(Jingui Yaolue,AD.～ 220)” in Han dynasty,and is to treat body fluid disorders and dyslipidemia initially.Although many bioactive monomers from Zexie and Baizhu have been discovered to improve lipid disorders,glucose metabolism,and inflammation,there is limited research on aqueous ZXBZ decoction,the original clinical formulation.Considering ZXBZ decoction is the most original prescription without systemic study,this research determined to verify the efficacy in NAFLD,clarify the mechanisms,and screen the valid components.Firstly,three different NAFLD mouse models were used.(1)Gubar-Amylin NASH(GAN)-induced NAFL mouse model,which is close to the pathogenesis in obesity population with NAFL;(2)High Fat High Cholesterol Diet(HFHCD)-induced db/db mouse NASH model characterized by hyperglycemia and insulin resistance;(3)GAN diet combined with CCl4 induced NASH mouse model with significant inflammation and fibrosis.We explored that ZXBZ decoction could protect liver function and improve lipid metabolism significantly in 17 week-GAN induced NAFL mouse model.But in two NASH models,ZXBZ decoction could only decrease the serum hepatic enzymes in the progress from NAFL to NASH.Fortunately,the serum lipid levels were still decreased significantly in ZXBZ decoction treated groups,and it could also improve the oral glucose tolerance.Then we investigated the underlying mechanisms for the further development and exploration of ZXBZ decoction based on transcriptomics analyses,network pharmacology,and molecular biology experiment.We discovered ZXBZ decoction could activate two energy sensors,Sirt1 and AMPK,hence,change-related genes involved in lipogenesis(SREBP-1c,Ch REBP,HMGCR),lipid catabolism(CPT-1α,LPL),lipid transport(LXRα)and bile acid metabolism(CYP7A1,CYP17A1)and regulate the activity of lipid metabolizing enzymes(ACC,FASN),subsequently improved the lipid disorders in NAFL mice.Besides,PA-stimulated Hep G2 cells,AMPK,and Sirt1 selective inhibitors were used to illuminate the mechanisms in vitro.We found ZXBZ decoction could activate AMPK and Sirt1 separately and independently,either inhibitor could not affect the ZXBZ decoction’s activation towards the other one.We also screened 3 anti-steatosis ZXBZ ingredients out of 13 components based on FFAs induced Hep G2 cells lipid accrual models.In conclusion,the pharmacodynamical research,pharmacological studies,and bioactive ingredients screening of ZXBZ decoction could help promote the modern development and application of Traditional Chinese Medicine.