Protective Effects Of Madecassoside In Acute Liver Failure In Mice Induced By LPS/D-GalN

Objective: Madecassoside(MA),a triterpenoid saponin isolated from Centella asiatica,exerts various pharmacological activities including antioxidative and anti-inflammatory actions.The aim of this study was to explore the protective effect of MA in the treatment of lipopolysaccharide(LPS)and Dgalactosamine(D-Gal N)-induced acute liver failure(ALF)in mice.Method: A mouse model of acute hepatic injury induced by LPS/D-GalN was constructed.Mice were randomly divided into control group,LPS/D-GalN group,silibinin group,and MA(10,20,40 mg/kg)group,they were administrated once daily for 10 days to observe the physiological condition of mice continuously.On day 10,all mice were injected intraperitoneally with LPS 10 ?g/kg and DGalN 700 mg/kg dissolved in physiological saline except the control group,and blood and liver samples were collected 8 hours later.The liver function parameters of mice were detected,H&E staining was used to stain the liver tissues and observe the cell structure and pathological changes,Western blotting was used to detect The protein expression levels of p38 MAPK and p-p38 MAPK,NF-?B and p-NF-?B,COX-2,i NOS,Nrf2 and HO-1 in liver tissue,and the mRNA expression levels of TNF-?,IL-1?,IL-6,i NOS,COX-2,SOD1,CAT and GPx1 in liver tissue were detected by real-time fluorescent quantitative PCR.The levels of IL-1?,IL-6,TNF-?,SOD,CAT and GPx in liver tissue were detected by ELISA.Result: Compared with the control group,hepatic samples from LPS/DGalN-treated mice had nuclear fragmentation,cytoplasm condensation,fragmented desmosome complexes,and cells separated from their neighbors in liver pathological sections,and the levels of ALT and AST increased significantly(P<0.01).Compared to the LPS/D-GalN group,administration of MA at doses of 20 and 40 mg/kg significantly reduced the levels of ALT and AST(P<0.05).Among them,according to pathological section imaging.The administration dose of MA at 20,40 mg/kg significantly improved the histopathological state of acute liver failure induced by LPS/D-GalN,and There was a significant inhibitory effect in the phosphorylation progresses of p38 MAPK and NF-?B after administration of MA at 20 and 40 mg/kg(P<0.01),and MA at 10 mg/kg couldn't ameliorate the phosphorylation of NF-?B significantly(P>0.05).Compared with LPS/D-GalN group,MA at 20 and 40 mg/kg had significant inhibitory effect on the expression of iNOS and COX-2(P<0.01).At the doses of 20 and 40 mg/kg in MA,SOD,CAT and GPx showed an upward trend in the antioxidant system in mice liver tissue,which was significantly different from the LPS/D-Gal N group(P<0.01).At the same time,MA promoted the expression of Nrf2 and HO-1 after exposed to LPS/D-GalN in mice,and there was a significant increase in the dose of 20 and 40 mg/kg compared with LPS/D-GalN group.Compared with LPS/D-GalN group,the inflammatory cytokines in liver tissue,when the dose of deliverociorin was 20 or 40 mg/kg,was a significant downtrend including the levels of TNF-?,IL-1? and IL-6 in liver tissue.In the real-time fluorescence quantitative PCR experiment,compared to the LPS/D-Gal N group,administration of MA at a dose of 20 mg/kg or 40 mg/kg significantly reduced the mRNA levels of TNF-?,IL-1?,and IL-6,iNOS and COX-2 and close to normal(P<0.01),and they also stimulated their antioxidant enzymes to further activations,expressed higher levels of SOD1,CAT and GPx1 in mRNA than LPS/D-GalN group(P<0.01).Conclusion: Our study demonstrated that MA provided significant protection against ALF induced by LPS/D-GalN in mice.These effects may be mediated by blocking inflammation through suppression of the p38 MAPK and NF-?B pathways,caused by the reduction in TNF-?,IL-1?,and IL-6,and by preventing oxidative damage through stimulation of anti-oxidative enzymes,Nrf2 and HO-1,which kept AST and ALT at normal levels.Thus,our data suggest that MA is beneficial in preventing ALF induced by LPS/D-GalN in mice.