The Effects Of Mongolian Drug Digeda-4decoction And Active Ingredients On Protecting Acute Liver Injury In Vivo And In Vitro

Objective:To observe the effect of Mongolian drug Degeda-4 decoction on protecting acute liver injury in vivo and in vitro,and to analyze its transitional elements in vivo and identify the active ingredients as the hepatoprotective effect of Degeda-4 decoction.Methods:(1)The animal model of acute liver injury in rats was established by D-galactosamine(D-GlaN)and carbon tetrachloride(CC14).The protective effect of aqueous extract of Degeda-4 decoction on acute liver injury in rats was evaluated on serumal and pathological indicators.(2)The fingerprint of Mongolian drug Degeda-4 decoction was established by HPLC.(3)Transitional elements of Degeda-4 decoction in serum,liver,lung,heart,kidney,spleen,urine were isolated and identified by LC/MS and NMR.(4)The maximum safe concentration of drug was determined by cytotoxicity assays in Hela cell line.The effects of serum and aqueous extract of Mongolian drug Degeda-4 decoction,swertiamarin,beta sitosterol,palmitic acid,geniposide and picroside Ⅱ on protecting acute liver injury established by D-GlaN and CCl4(HSC-T6)were observed in vitro.Results:(1)High,medium and low dose groups of Digeda-4 decoction could significantly reduce serum glutamic oxaloacetic transaminase(AST)and alanine aminotransferase(ALT)levels(P<0.05)in acute liver injury rat model induced by D-GlaN and CCl4.Medium dose group of Digeda-4 decoction could significantly reduce serum alkaline phosphatase(ALP)level(P<0.05).(2)The fingerprint of Degeda-4 decoction with 21 common peaks was established by study of Lomatogonium,Rhizoma picrorhizae,CapeJasmine and Herba dianthi.(3)20 transitional components of Digeda-4 decoction were detected in vivo,including 9 original components and 11 metabolites.The 9 prototype components are 4-0-β-glucopyranosylacetophenone,androsin,swertiamarin,beta sitosterol,palmitic acid and geniposide,Picroside Ⅱ.Metabolite structure is inferred by fragment ions obtained by MS and two stage mass spectrometry and UV spectral data.(4)Digeda-4 decoction serum,aqueous extract of Digeda-4 decoction,swertiamarin,beta sitosterol,palmitic acid,geniposide and Picroside Ⅱ in safe concentration showed varying degrees of protection of damage in HSC-T6 cells induced by D-GlaN and CC14.The AST,ALT,transforming growth factor beta 1(TGF-beta 1),collagen-1(Col-1)and alpha smooth muscle actin(alpha-SMA)of cell culture medium were decreased under the influence of drugs,compared with the control group(P<0.05)Conclusion:(1)Mongolian medicine Degeda-4 decoction has obvious protective effect on acute liver injury induced by D-GlaN and CCl4.(2)Transitional composite on by LC/MS and NMR spectral analysis from the Degeda-4 decoction may be the main Pharmaco dynamic material basis of’ Mongolian medicine Degeda-4 decoction as a liver protecting agent.