| Objective:To investigate the effects of Danshensu on transforming growth factorβ1(TGFβ1)/Smads/ extracellular signal-regulated kinase (ERK) signaling pathways and the interaction of the pathways in hepatic stellate cell(HSC).Method:The rat HSCs were isolated in situ-liver recirculation and cultured in vitro. MTT colorimetric assay choosed the optimal concentrations of Dashensu;HSC induced by TGFβ1 were effected by the optimal concentrations of Danshensu and ERK Blocking Agent (PD98059).The profiferation of HSC in each group was measured by Cell Counting Kit-8(CCK-8). The expression ofα-smooth muscle sctin(α-SMA)and collagen I,III in HSC of each group was detected by the immunocytochemical assay. Smad2,3,7 mRNA in HSC of each group was detected by RT-PCR.The level of phosphorylation of Smad2/3,ERK1/2 protein and Smad7 protein was detected by Western blotting.Result:.At the concentration of 0.0625 to 0.25mmol/L,Danshensu significantly inhibited the proliferation of HSC. PD98059(100μmol/L) inhibited the profiferation of HSC induced by TGFβ1,Danshensu inhibited the profiferation of HSC induced by TGFβ1 in a dose-dependent manner(P<0.01); TGFβ1 could increase the expression level ofα-SMA and collagen I,III in HSC (P<0.01),but PD98059 and Danshensu could decrease the level(P<0.01); PD98059 depressed the expression level of Smad2,Smad3,Smad7 mRNA (P<0.01,P<0.05).Danshensu down-regulated the level of Smad2,3 mRNA,while up-regulated the level of Smad7 mRNA(P<0.01);PD98059 and Danshensu depressed the contents of phosphorylated Smad2/3 and ERK1/2 proteins(P<0.01),while they had different effects on the expressions of Smad7 protein(P<0.01).CONCLUSION:The TGFβ1 up-regulated the expressions of Smad2,3 mRNA and phosphorylated Smad2/3 proteins to induce the profiferation and activation of HSC,and to induce the synthesis of collagen in HSC,ERK pathway could participated the expressions of Smad genes and phosphorylated proteins in HSC induced by TGFβ1;Danshensu could inhibited the signaling pathways of TGFβ/Smad/ERK in hepatic fibrosis . |
