Isolation And Identification Of Chemical Constituents From Rhinacanthus Nasutus(L.)Kurz And Study On Its Effect And Mechanism On Non Alcoholic Fatty Liver Disease

Based on the traditional efficacy,modern pharmacology,and lipid-lowering activity screening results of Rhinacanthus nasutus(L.)Kurz,this article conducted an in-depth research on the chemical components of Rhinacanthus nasutus and its effect on improving NAFLD based on active guidance.Rhinacanthus nasutus is an upright perennial herb of the Acanthaceae family,widely distributed in Yunnan,Guangxi,Guangdong,and Southeast Asia.As a national medicine,it has a medicinal history dating back hundreds of years and is mainly used in Zhuang and Yao mountain areas for the treatment of acne,scabies,snake bites,and chronic hepatitis.This plant contains a variety of chemical components and has significant anti-tumor,diabetes-treating,lipid-lowering,and liver-protecting pharmacological effects.To further clarify the material basis of Rhinacanthus nasutus for reducing lipids and protecting the liver,this study used activity-directed separation methods and multiple chromatography separation techniques to separate and purify the petroleum ether and ethyl acetate fractions of Rhinacanthus nasutus.The structures of the compounds were identified using IR,UV,NMR,and MS techniques;a total of 36 compounds were identified,including 9 flavonoids,10 naphthoquinones,17 phenylephrine,lignans,and small molecular phenols.Among them,14 compounds were isolated for the first time from Rhinacanthus nasutus.Using the oleic acid-induced high-fat HepG2 cell model,22 compounds were screened for their antitumor and lipidlowering activities.Their cytotoxic activity was time-and concentration-dependent.The results of the lipid-lowering activity showed that the 15 compounds had varying degrees of lipid clearance,with the lipid clearance rates of RC,RT,and RN on the HepG2 high-fat model being as high as 50% or more.Among them,the effects of RC,RT,and RN on reducing TC and TG were better than those of the lovastatin positive group,indicating that they had better preventive and therapeutic effects on fatty liver.The RRE of Rhinacanthus nasutus has the effect of improving liver lipid deposition and lipid metabolism disorders in HFD-induced NAFLD mouse models,and its mechanism may be related to the improvement of intestinal microflora diversity and microflora structure of NAFLD in response to the RRE.In addition,a model of NAFLD was established using HFD induction.Based on nontargeted metabolomics,the effects of RC on NAFLD liver disease and its potential molecular biological mechanisms were studied.The results showed that RC can significantly improve liver lipid metabolism disorder,fat deposition,and inflammatory damage in HFD-induced NAFLD models.Citric acid,iscitric acid and oxoglutaric acid can be used as potential metabolic markers in the clinical serum of HFD-induced NAFLD.The mechanism of RC’s upregulation of lipid-related metabolites may be based on the AMPK-SIRT1-NF-κB signal pathway-mediated oxidative stress and inflammatory protection.The mechanism of RC’s downregulation of organic acid metabolites may be related to the regulation of the intestinal microflora structure of the host.The mechanism of action of RC may be that it regulates that the abundance of Proteobacteria,Fusobacteriata,and Bacteroides by regulating the composition of the intestinal microbial community structure,The mechanism of the effect of RC on lipid metabolism disorder in NAFLD mice may be via regulating the composition of the intestinal microbial community structure,upregulating the abundance of Proteobacteria,Fusobacteriota and downregulating the abundance of Bacteroides.Through regulating the lipid metabolism disorder and unsaturated fatty acid synthesis in mice,RC downregulates the abundance of L-Tryptophan,trehalose,iscitric acid,and oxoglutaric acid in serum,and detegulating the abundance of 5-HETE,PE(22:5(7Z,10 Z,13Z,16 Z,19Z)/20:1(11Z))and alpha-Dimerphelic acid are involved in regulating the AMPK-SIRT1-NF-κB signal pathway,which mediates lipid metabolism disorders and oxidative stress caused by inflammatory damage,while reducing the expression of lipid synthesis functional proteins SREBP1 c and FAS,thereby improving lipid metabolism disorders and inflammatory damage in the pathological process of NAFLD,and reducing liver fat deposition.