Effects Of YPF-P On The Proliferation Of RAT HSC Which Reduced By PDGF-BB And ERK1 / 2 Signal Transduction Pathway

Objective:To investigate the inhibitory effect of YPF-P on proliferation,collagen typeⅠ,collagen typeⅢ,α-SMA,PDGF-BB,PDGFR-β,Phosphorylated extracellular signal-regulated kinase 1/2(P-ERK1/2)and c-fos expression of rat hepatic stellate cel(lHSC)stimulated by platelet derived growth factor-BB(PDGF-BB),and to elucidate the molecular mechanism of Danshensu against hepatic fibrosis.Methods:1. The influence of different consentrations of YPF-P on HSC proliferation was detected by MTT assay at different time, calculating the inhibitory rates to determine the YPF-P's best role time and role concentration .2. To investigate the effect of YPF-P on proliferation,collagen typeⅠ,collagen typeⅢandα-SMA mRNA expression of HSC stimulated by PDGF-BB by RT-PCR.3. To investigate the effect of YPF-P on PDGF-BB,PDGFR-βmRNA expression of HSC stimulated by PDGF-BB by RT-PCR.4. To detect the protein expression of ERK1/2 and P-ERK1/2 under different interventions on HSC stimulated by PDGF-BB by Western blotting. 5. To investigate the effect of YPF-P on c-fos and c-jun mRNA expression of HSC stimulated by PDGF-BB by RT-PCR.Results:1. The effects of YPF-P on proliferation of HSC-T6 induced by PDGF-BB The model group stimulated by exogenous stimulating factor PDGF-BB (10ng/ml) compared with the normal control group of HSC-T6 without exogenous stimulating factor ,which have almost no change at the 24h, the difference don't have statistically significant,but the proliferation was obvious at 48,72 h, the difference has statistically significant, and the most obvious proliferation was at 48h; the role of 12.5,25,50,100,200 mg / L of the YPF-P group compared with the group stimulated by PDGF-BB ,the proliferation of PDGF-BB-induced HSC all can be inhibited,and With the concentration of YPF-P enhanced the inhibitory enhanced. In addition to 12.5mg / L of the YPF-P was not obvious, the other inhibition of HSC groups were all obvious.2. The effect of YPF-P on collagen typeⅠ,collagen typeⅢandα-SMA mRNA expression of HSC stimulated by PDGF-BB In the normal control group,the expression of collagenⅠ,Ⅲcollagen andα-SMA mRNA of HSC-T6 was relatively low,butthe expression of collagen typeⅠ,collagen typeⅢandα-SMA mRNA of the model group was significantly higher than the normal group (P <0.01, P <0.05); 25,50,100 mg / L of the YPF-P group could significantly inhibit the increase of collagenⅠ,Ⅲcollagen andα-SMA mRNA level, and with the YPF-P concentration increased inhibition enhanced.3. The effect of YPF-P on PDGF-BB and PDGFR-βmRNA expression of HSC stimulated by PDGF-BB In the normal control group,the expression of PDGF-BB and PDGFR-βmRNA of HSC-T6 was relatively low,butthe expression of PDGF-BB and PDGFR-βmRNA of the model group was significantly higher than the normal group (P <0.01, P <0.05); 25,50,100 mg / L of the YPF-P group could significantly inhibit the increase of PDGF-BB and PDGFR-βmRNA level, and with the YPF-P concentration increased inhibition enhanced.4. The effect of YPF-P on the protein expression of ERK1/2 and P-ERK1/2 under different interventions of HSC stimulated by PDGF-BB PDGF-BB induced HSC-T6 cells group compared with the normal group,the expression of phosphorylation of ERK1/2(p-ERK1/2) was increased significantly, U0126 which is the inhibitor of ERK pathway and YPF-P the expression of p-ERK1/2 of PDGF-induced HSC-T6 cells, and the combination of U012 6 and YPF-P can reduce more obviouly.5. The effect of YPF-P on c-fos and c-jun mRNA expression of HSC stimulated by PDGF-BBIn the normal control group,the expression of c-fos and c-jun mRNA of HSC-T6 was relatively low,butthe expression of c-fos and c-jun mRNA of the model group was significantly higher than the normal group (P <0.01, P <0.05); 25,50,100 mg / L of the YPF-P group could significantly inhibit the increase of c-fos and c-jun mRNA level, and with the YPF-P concentration increased inhibition enhanced.Conclusion:YPF-P could inhibit HSC proliferation collagen typeⅠ,collagen typeⅢ,α-SMA,PDGF-BB,PDGFR-β,c-fos and c-jun significantly,and its mechanism may be related to the inhibition of ERK1 / 2 signal transduction pathway which induced byPDGF-BB .