大鼠肝星状细胞活化过程中过氧化物酶体增殖因子活化受体γ表达的动态变化

Dynamic change of peroxisome proliferator-activated receptor γ expression during rat hepatic stellate cells activation

目的 研究大鼠肝星状细胞（HSC）活化过程中过氧化物酶体增殖因子活化受体γ（PPARγ）表达的动态变化。方法 采用肝脏原位灌流酶消化，Nyeodenz密度梯度离心方法体外分离大鼠HSC，分别采用半定量RT-PCR和Western blot检测基因和蛋白表达水平。结果 HSC体外培养过程中，出现与体内肝纤维化发生过程中相似的活化过程。在静止、中间活化、活化的HSC中，PPARγ达水平不断下降。结论PPARγ表达水平随着HSC活化程度的增加而不断下降，可能在维持HSC静止表型方面发挥重要作用。

Objective To stuely the dynamic change of peroxisome proliferatoroactivated receptor γ（PPARγ） expression during rat hepatic stellate cells （HSC） activation. Method Rat HSC were isolated from normal Sprague Dawley rats through in situ perfusion of liver with Pronase E and density-gradient centrifugation with Nycodenz. Total RNA was extracted by TRizol reagent and gene levels were determined by semi-quantitative RT-PCR. Total cellular and nuclear protein were isolated respectively, and separated by 10% sodium dodecylsnifate polyacrylamide gel electrophoresis. Result As HSCs phenotype underwent activation gradually with the culture duration prolongation, PPARγnuclear expression level also decreased not only at transcriptional level but also at traslation level. Conclusion PPARγ expression decreased with HSC activation extent and it plays important role in maintaining of HSC silent phenotype.