Study On The Effects And Significance Of Abstract Of Periplaneta Americana On Mitogen Activated Protein Kinsae Signal Pathway In Rat Hepatic Stellate Cells In Vitro

Objective:To study the effcets of Abstract of Periplaneta Americana (APA) in rat hepatic stellate cells (HSC-T6) expression of P-P38, P-ERK, P-JNK important signaling molecules in MAPK signaling pathway. And to explore the effects of MAPK signaling pathway inhibitors on the rat hepatic stellate cells (HSC-T6) proliferation, providing molecular mechanism of Abstract of Periplaneta Americana (APA) on anti hepatic fibrosis.Methods:1 Rat HSC line (HSC-T6) were incubated with different concentrations (0.5mg/mL、1mg/mL、2mg/mL、4mg/mL、mg/mL、8mg/mL、10mg/mL 16mg/mL) of APA and or without APA for 24h, Cells proliferation was examined by MTT.2 The cultured HSC-T6 were divided into five experimental groups:Control group, different concentrations (2mg/mL,4mg/mL,6mg/mL,8mg/mL) of APA group, after half an hour, the P38, P-P38, ERK1/2, p-ERK1/2,JNK, P-JNK expression levels were detected by western blot..3 Rat HSC line (HSC-T6) were incubated with different concentrations of the P38MAPK inhibitors SB203580 (O.1uM,0.5uM,2.5uM),ERK1/2 inhibitors PD98059(0.4uM,2uM, lOuM), JNK inhibitors SP600125 (0.008uM,0.04uM, 0.2uM)and or without them for 24h, Cells proliferation was examined by MTT.Results:1 MTT assay indicated:different concentrations of APA intervention HSC-T6 cell, proliferation in the experimental group were lower than the control group, the difference was statistically significant (P<0.01); when the concentrations of APA between 0.5-4mg/ml, with the increase of drug concentrations, cell proliferation inhibition rate gradually increased in a dose dependent. And when the concentration of APA between 4-16 mg/ml, with the increase of drug concentrations, cell proliferation inhibition rate gradually decreased in a dose dependent.2 The results of western blotting assay indicated:After APA intervention HSC-T6 half an hour, the expression of P38, P-P38, ERK1/2, p-ERK1/2,JNK, P-JNK levels were in varying degrees decreased. Different concentrations of APA could significantly inhibit the expression of p-JNK and in the experimental group it was lower than the control group, the difference was statistically significant (P<0.01), and showed a good in a dose dependent.And the 2mg/mL of APA group the expression of p-ERK and p-p38 were not effected and compared to the control group, the difference was statistically significant (P<0.01), Only when the concentration of APA is equal or more than 4mg/ml can significantly inhibit the expression of P-P38 and P-ERK, and compared to the control group, the difference was statistically significant (P<0.01), but it did not show in a dose dependent.3 MTT assay indicated:different concentrations of MAPK (P38, ERK1/2andJNK) signal pathway inhibitors intervention HSC-T6, proliferation in the experimental group were lower than the control group, the difference was statistically significant (P<0.01). and with the increase of inhibitors concentration, the cell proliferation inhibition rate decreased gradually, and showed a good in a dose dependent.Conclusion:1 The Abstract of Periplaneta Americana can significantly inhibit the proliferation of HSC-T6 in a dose-dependent; And high concentrations of APA have some protective effect on HSC-T6 cells.2 The Abstract of Periplaneta Americana could inhibit the expression of P-P38, P-ERK1/2 and P-JNK of HSC-T6.The mechanism of Anti-hepatic fibrosis may be related to the MAPK signal pathway.3 Different signal pathway inhibitor of MAPK intervention HSC-T6, the proliferation of cell in varying degrees decreased.