Genistein对大鼠肝脏星状细胞激活的抑制作用

Effect of Genistein on Inhibiting the Activation of Hepatic Stellate Cells

目的 探索酪氨酸蛋白激酶抑制剂 Genistein对肝脏星状细胞 (HSC)激活的抑制作用。方法 采用链霉蛋白酶、胶原酶原位灌注及 Nycodenz一步密度梯度离心法分离大鼠 HSC,并进行体外培养使之激活。用四唑盐比色法 (MTT法 )检测 Genistein作用 2 4、4 8和 72小时对 HSC增殖的抑制作用 ;用激光扫描共聚焦显微镜(L SCM)观察 Genistein作用后 HSC的 α-平滑肌肌动蛋白 (α- SMA,HSC激活的标志之一 )表达的变化。结果 MTT检测显示 ,10 - 5～ 10 - 9mol/ L 浓度的 Genistein作用 2 4、4 8和 72小时对 HSC增殖有抑制作用 ,并以 10 - 7mol/ L 的 Genistein作用 4 8小时的抑制作用最明显。 L SCM观察显示 ,10 - 6和 10 - 7mol/ L 的 Genistein作用 4 8小时可使 α- SMA表达明显减弱。结论 一定浓度的 Genistein能明显抑制 HSC的活化和增殖 。

Objective To explore the effect of Genistein, a tyrosine protein kinase inhibitor, on inhibiting the activation of hepatic stellate cells (HSC). Methods The HSC from Wistar rats were prepared by in situ perfusion and single step density Nycodenz gradient and activated by culturing. The inhibitive effect of Genistein on the proliferation of HSC was examined using tetrazolium blue test after 24, 48, and 72 hours of interaction. The α smooth muscle actin (α SMA), one of the markers of the activated HSC, was evaluated by the laser scanning confocal microscope. Results Genistein in the concentrations of 10 -5 10 -9 mol/L inhibited the proliferation of HSC, with a maximal effect after 48 hours interaction and at the concentration of 10 -7 mol/L. Conclusion Genistein inhibits the proliferation and activation of HSC and may become potential drug in preventing liver fibrosis in the future.