Preliminary Study On Anti-hepatic Fibrosis Effect And Material Basis Of Swertia Chirayita

Hepatic fibrosis is a common response to various chronic hepatic injuries,characterized by the deposition of extracellular matrix(ECM).Liver fibrosis itself causes no symptoms,but the advanced fibrosis can lead to irreversible cirrhosis or hepatocellular carcinoma,and finally often requires liver transplantation.Previous reports have indicated that liver fibrosis is potentially reversible,so the early diagnosis and intervention of this disease have important clinical implications.Until now,numerous works have centered on gene expression,protein function,as well as conventional pathophysiological studies on the liver fibrosis.The mechanisms for liver fibrosis have been dramatically elucidated but are still not fully understood.Since liver fibrosis is shown to be reversible,efforts should be made to search for effective therapeutic approaches.Unfortunately,no established anti-fibrotic therapies are currently available.Traditional Chinese medicine(TCM),owing to its good efficacy and few side effects,has displayed unique features in the treatment of liver diseases for thousands of years.In recent years,tibetan medicine,a kind of TCM,have attracted increased attention in treatment of diseases due to its excellent effect in the treatment of liver diseases.It is an inevitable trend to strengthen the basic research and development of Tibetan medicine,and to develop a better activity of anti fibrosis drugs.Swertia chirayita is the whole herb of Swertia chirayita(Roxb.ex Flemi)Karsten,which has a long history of clinical application,and the significant characteristics of the Tibetan Medicine.The previous studies showed that Swertia chirayita could protecting the liver.However,whether the protective effect of liver can play a further role in anti-fibrosis is still unknown,and the material basis has not been systemically studied,which limits the development of Swertia chirayita.In this work,we investigated the anti-fibrosis effect of Swertia chirayita in vitro and in vivo,as well as material basis of this herb,aiming to elucidating the understanding the potential mechanism and material basis of Swertia chirayita.1.Literature research The paper summerize the mechanism of hepatic fibrosis and the research progress of TCMin the treatment of hepatic fibrosis.The development of metabonomics was systemically reviewed.And constituents from Swertia chirayita were summerzied,including physicochemical properties,pharmacological and other basic research content.2.Hepatic fibrosis inhibitory effect of Swertia hirayita in vivoTo observe the hepatic fibrosis inhibitory effects of Swertia Chirayita on carbon tetrachloride(CCl4)-induced mouse model of liver fibrosis.Liver fibrosis was induced in rats using CCl4 by intraperitoneal injection for 7 weeks.32 male SD rats were randomly divided into normal group,model group,Swertia chirayita group,positive control group.The test group was given the corresponding drugs once a day for 8 weeks.At the end of the experiment,MASSON staining evaluated the extent of liver fibrosis.Serum HA,LN,Pc Ⅲ and ColIV level were measured by ELISA.And serum ALT,AST,ALP,TBIL were determined by enzymatic methods.Compared with normal group,it was showed that the MASSON staining of model group signficant changed,serum HA,LN,PC Ⅲ,ColIV increased significantly(p<0.01).AST,ALT,ALP and T-BIL levels were also increased significantly(p<0.05).Compared with model group,the degree of MASSON staining of Swertia chirayita group decreased,as well as the levels of HA,LN,PC and CIV(p<0.01),ALT,T-BIL levels were significantly lower(p<0.05),the level of AST,ALP show a downward trend.The result showed that Swertia Chirayita has had an anti-fibrosis effect in the CCl4-induced liver fibrosis mode.3.Hepatic fibrosis inhibitory effect of Swertia chirayita in vitroTo investigate the anti-fibrosis mechanism of Swertia chirayita,used MTT analysis it’s effects on the proliferation and apoptosis of rat hepatic stellate cells.The experimental results suggested that Swertia chirayita can inhibit the proliferation of HSC,and in the dependent manner of concentration.This indicated that the mechanism of Swertia chirayita for treatment of hepatic fibrosis may be related by the inhibition of the proliferation of hepatic stellate cells.4.Serum metabolomic characterization of liver fibrosis in rats and anti-fibrotic effects of Swertia chirayitaIn this work,an UPLC-TOF-MS-based serum metabolomic study was performed to investigate the potential biomarkers and pathway of liver fibrosis in rats,as well as the anti-fibrotic effects of Swertia chirayita.After multivariate statistical analysis,a clear separation of the model and control groups was achieved,and the Swertia chirayita group exhibited a tendency to restore to the controls.Seventeen significantly changed fibrosis-related metabolites were identified.The therapeutic effects of Swertia chirayita may be attributed to tryptophan metabolism,bile acid metabolism etc metabolic pathways in liver fibrosis.These findings will contribute to increased understanding of liver fibrosis and the anti-fibrotic mechanisms of Swertia chirayita.5.Rapid screening and structural characterization of constituents from Swertia chirayita by UPLC-Q-TOF-MSBy UPLC-Q-TOF-MS anlysis,the main components from Swertia chirayita were identified and characterized.The analysis was performed on an ACQUITY HSS T3 reverse phase column(2.1 mm×100 mm,1.8 μm).0.1%formic acid in ACN and 0.1%formic acid in water was used as mobile phase for gradient elution.The flow rate was 0.4 mL/min.Mass spectrometry was applied for the qualitative analysis under positive and negative ion modes and ESI ion source was used for mass spectra.24 compounds were identified or tentatively assigned from Swertia chirayita,including 6 iridoids,11 xanthones,2 flavonoids,2 isocoumarin,2 phenols and 1 terpenoids.Among the compounds analyzed,5 compounds were unambiguously identified by comparing their retention times,MS/MS spectra with that of reference compounds,while others were tentatively assigned by comparing their retention behaviors,empirical molecular formula and proposed fragmentations with that in literatures,And five of these compounds were characterized from this specie for the first time.In summary,anti-hepatic fibrosis activity of Swertia chirayita was proved in vitro and in vivo,as well as the mechanism of anti-hepatic fibrosis was confirmd by metabolomics.And the chemical profile was revealed by UHPLC-QTOF-MS.The study will provided scientific basis to rationalize and develop Swertia chirayita.