Effect Of Danshengsu On ERK1/2 Signal Transduction Pathway In Rat Hepatic Stellate Cell

Objectives:To investigate the inhibitory effect of Danshengsu on proliferation,collagen typeⅠsynthesis and Phosphorylated extracellular signal- regulated kinase 1/2 (P-ERK1/2)expression of rat hepatic stellate cell ( HSC ) stimulated by platelet derived growth factor-BB (PDGF-BB),and to elucidate the molecular mechanism of Danshensu against hepatic fibrosis。Methods :Rat HSC was isolated by an improved collagenase-pronase digested method and identified byα-SMA immunocytochemical stain;MTT colorimetric assay was used to evaluate Danshensu's inhibitory effect on proliferation of HSC stimulated by PDGF-BB。Inhibitory effect of Danshensu on Collagen typeⅠsynthesis of HSC stimulated by PDGF-BB was detected by Immunocytochemical stain;Inhibitory effect of Danshensu on P-ERK1/2 expression in HSC stimulated by PDGF-BB was detected by Western blot。Results:1.The yield of HSC was about3.5×10~7 per rat;Survival rate of HSC exceed 98% and purity of the second passage HSC exceed 99%。2. Compared with normal group , proliferation of HSC stimulated by PDGF-BB was significantly enhanced (P<0.01) ; Danshensu had an inhibiting effect on the proliferation of HSC stimulated by PDGF-BB(P<0.05) ; The inhibtory effect was concerned with it , s dose(P<0.01)。3. PDGF-BB could stimulate collagen typeⅠsynthesis in HSC(P<0.01);Danshensu had a inhibtory effect on PDGF-BB induced collagen typeⅠsynthesis of HSC in a dose-dependent manner (P<0.05)。4 P-ERK1/2 expression of HSC stimulated by PDGF-BB was reinforced; Danshensu could downregulate P-ERK1/2 expression of HSC ; The inhibtory effect was concerned with it's dose。Conclusions : 1. Primary cultured rat HSC was an important experimental object to investigate liver fibrosis in vitro.2. Danshensu could inhibit proliferation and collagenⅠsynthesis of HSC stimulated by PDGF-BB, the mechanism of which may be Danshensu,s inhibitory effect on ERK1/2 signal transduction pathway。...