HSP47 Inhibitor IPA1010 Improves The Mechanism Of High-fat-induced Non-alcoholic Steatohepatitis (NASH) In Gerbils

Objective:To establish stable and replicable models of nonalcoholic steatohepatitis(NASH)gerbils and evaluating heat shock protein 47(HSP47)inhibitor IPA1010’s regulatory mechanism on hepatic pathological condition and hedgehog pathway-assosiated proteins’expression.Methods:health male gerbils(50-60 g)were fed with a commercially available standard diet for 1 week in the experimental environment before the experiments,sixty gerbils were divided randomly into six groups:the normal control group(NC),model control group(MC),simvastatin treated group(PC),low,medial and high dose of IPA1010 treated groups.Each group had 10 gerbils.The gerbils were fed with high fat diet(HFD)for 12 weeks to induce NASH.From 9th week to 12th week,the treated drugs were given daily to each animal by oral administration,and saline to NC and MC groups,respectively.On the last day of 12th week,gerbils were sacrificed and blood samples were collected and centrifuged to obtain serum.The levels of serum insulin(INS),hyaluronic acid(HA),hydroxyproline(Hyp),procollagen type III(PC III),procollagen type IV(PC IV),transforming growth factor-β1(TGF-β1)and interleukin-1β(IL-1β)were detected by enzyme-linked immuno sorbent assay(ELISA).The levels of serum alanine aminotransferase(ALT),aspartate aminotransferase(AST),total cholesterol(TC)and triglycerides(TG)were detected by chemical chromatometry.The number of CD4+T,CD8+T and NK cells were determined by immunohistochemistry and the expression of HSP47,TGF-β1,nuclear factor-KB(NF-κB)p65,sonic hedgehog(SHH)and GLI3 were detected by western-blot.Results:We established the model of NASH through 12 weeks HFD feeding.Compared with the MC group,IPA1010 could improve the physiological status of liver and regulate blood lipids through reducing serum levels of ALT,AST,TC and TG.Improve the status of liver fibrosis through reducing serum levels of PC Ⅲ,PC Ⅳ,TGF-β1 and IL-1β.The infiltration of CD4+T and NK cells were alleviated after treated with IPA1010 compared with MC group and the expressions of HSP47,TGF-β1,NF-κB p65 and SHH were down regulated while GLI3 up regulated obviously.Conclusions:The HSP47 inhibitor IPA1010 improved the liver status which caused by HFD,clearly.It could dietary improve the function of liver,mitigation of fibrosis,promote immuno-microenvironment recovery,and its mechanism of improving the status of NASH might be its inhibition effect of HSP47 to suppress the hedgehog signal pathway,thus downstream of the inflammatory cytokines and related proteins’ expression were suppressed.