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目的 观察肿瘤生长因子 β1(TGFβ1)及血小板衍生生长因子 (PDGF)对大鼠肝窦内皮细胞 (sinusoidalendothelialcell,SEC)整合素α6β1表达及黏着斑激酶 (focaladhesionkinase ,FAK)活性的影响。方法 用胶原酶原位灌注、Percoll不连续密度梯度离心法分离大鼠SEC ,并进行体外培养。采用细胞 ELISA和免疫沉淀 蛋白质酪氨酸激酶活性测定法 ,分别观察TGFβ1及PDGF对SEC表面整合素α6β1表达及FAK活性的影响。结果 经TGFβ1及PDGF作用 2 4h后 ,α6β1蛋白表达明显强于对照组(P <0 .0 5 ) ,且呈剂量依赖性。作用至 48h ,表达继续增强 ,细胞中FAK的活性也明显增高 (P <0 .0 5 ) ,虽于 48h后回落 ,但仍明显高于对照组。结论 TGFβ1及PDGF可促进SEC表面整合素α6β1的表达及FAK活性增高 ,可能是它们参与肝纤维化发生的机制之一。
Objective To investigate the effect of TGFβ1 and PDGF on the expression of integrin α6β1 and the activity of focal adhesion kinase (FAK) in rat sinusoidal endothelial cells (SEC). Methods In situ collagenase (collagenase) perfusion and Percoll discontinuous density gradient centrifugation were used to separate rat SEC and cultured in vitro. The effects of TGFβ1 and PDGF on the expression of integrin α6β1 and the activity of FAK on SEC were observed by ELISA and immunoprecipitation assay. Results After treated with TGFβ1 and PDGF for 24 hours, the expression of α6β1 protein was significantly stronger than that of the control group (P <0.05), and the dose-dependent manner. After 48h, the expression of FAK in cells continued to increase, and the activity of FAK in cells was also significantly increased (P <0.05), though it dropped after 48h but was still significantly higher than that of control group. Conclusion TGFβ1 and PDGF can promote the expression of integrin α6β1 on the surface of SEC and the increase of FAK activity, which may be one of their mechanisms involved in the pathogenesis of hepatic fibrosis.