摘要
目的探讨蛋白酶活化受体(proteinase-activated receptors,PARs)对大鼠肝组织中肝星状细胞(hepatic stellate cell,HSC)活化和增殖的调控及其在肝纤维化中的作用。方法应用改良的Friedman方法和Nycodenz一步密度梯度离心法分离和培养结扎胆管诱导的肝纤维化SD大鼠HSC。观察HSC形态学变化,MTT法观察细胞的增殖能力,Western blotting检测a-SMA、PAR1和PAR2在HSC的表达,同时设立正常对照组和假手术组。结果随着培养HSCs时间的延长,HSC的VitA和脂滴逐渐减少,甚至消失,HSC形态发生改变呈星状,细胞增殖活跃,而a-SMA、PAR1和PAR2随着培养时间延长其表达逐渐增强,原代培养14天达到高峰,a-SMA与PAR1和PAR2的表达呈一致关系。结论PARs可促进a-SMA的表达,PARs表达与HSC的活化和增殖相关,参与了肝纤维化的形成。
Objectives To investigate the role of proteinase-activated receptors（PARs）in regulation the activation and the proliferation of rat hepatic stellate cell （HSC） in the experimental model of hepatic fibrosis induced by bile duct ligation （BDL）. Methods To adopt the experimental model of hepatic fibrosis induced by BDL,and rat HSC were isolated from control rats, sham operation rats and rats that had undergone BDL according to a modification of the technique of Friedmen. Liver was perfused exvivo and digested with solutions containing protase E followed by collagenase Ia. The digested liver specimen was filtered, the resulting suspension hepatocytes was cleaned of and resuspended in a discontinuous Nycodenz gradient. Centrifugation yielded HSC in the top layer and a Kupffer/endothelial ceils fraction in the lower layer. HSCs were cultured on plastic plates in Dulbecco＇ s modified minimal essential medium containing 20% FCS. Expression of a-smooth muscle actin （α-SMA） and expression of proteinase-activated receptor 1 and 2 （PAR1, PAR2） were examined with immunocytochemistry methods. Results In the course of culture rat HSC, the vitamin A and lipid droplet decreased or disappeared gradually, the cell morphous changed to stellate cells and cells proliferation activated. The expression of PAR1 and PARs paralleled with the expression of α-SMA,and correlated with activation and proliferation of HSC. Conclusion The expression of PARs correlated with activation and proliferation of HSC, PARs might play an important role in the mechanisms of hepatic fibrosis.
出处
《胃肠病学和肝病学杂志》
CAS
2007年第4期357-360,共4页
Chinese Journal of Gastroenterology and Hepatology
基金
中国人民解放军第三军医大学中青年基金
No.200543
关键词
蛋白酶活化受体
肝星状细胞
细胞活化
细胞增殖
肝纤维化
Proteinase-activated receptors
Hepatic stellate cell
Cell activation
Cell proliferation
Hepatic fibrosis.
