Tanshinol induces proliferation,apoptosis and NF-?B activation in cultured rat hepatic stellate cells after IL-1? inducement / 第三军医大学学报

ABSTRACT

Objective To investigate the effects of tanshinol on the proliferation,apoptosis and NF-?B activation in rat hepatic stellate cells(HSCs) after IL-1? inducement,and to elucidate the anti-fibrotic molecular mechanisms of tanshinol.Methods The rat HSCs was isolated with collagenase in situ liver recirculation perfusion and cultured in vitro.The cells were divided into 5 groups normal control,IL-1? treatment group(10 ng/ml),and tanshinol group 1,2 and 3.The later 3 groups were pretreated with tanshinol at the concentrations of 0.062 5,0.125 and 0.25 mmol/L respectively followed by 10 ng/ml IL-1? treatment 1 h later.MTT colorimetric assay was used to detect the proliferation of HSCs.AO/EB immunoflurorescence microscopy and combination Annexin-V-FITC/PI double-labelimmunofluorescence with flow cytometer were employed to examine the apoptosis of HSCs.Synthesis and secretion of collagen Ⅲ were detected by the quantitative immunocytochemical assay and ELISA respectively.The amounts of cytoplasm p-I?B? and NF-?B p65,and nuclear NF-?B p65 in HSCs were determined by Western blotting.Immunocytochemical staining and Western blotting were used to observe nuclear translocation of NF-?B p65.Results IL-1? increased the proliferation of HSCs(P