反转录-聚合酶链反应法研究肝星状细胞与细胞间粘附分子-1表达的关系

Study on the relationship of expression of intercellular adhesion molecule1 and hepatic stellate cells by RTPCR

目的研究肝星状细胞（ＨＳＣ）与细胞间粘附分子１（ＩＣＡＭ１）表达的关系。方法用链酶蛋白酶和胶原酶原位灌流，Ｎｙｃｏｄｅｎｚ密度梯度离心分别分离正常大鼠及四氯化碳实验性大鼠肝纤维化模型中的ＨＳＣ，并进行体外培养，应用免疫组织化学方法和反转录聚合酶链反应（ＲＴＰＣＲ）技术分别观察不同培养时期的ＨＳＣ、正常及造模肝组织中ＨＳＣ的ＩＣＡＭ１的表达。结果正常大鼠新分离的ＨＳＣ中，ＩＣＡＭ１不表达；原代培养第１０天及传代培养第７天的ＨＳＣ表达ＩＣＡＭ１，且随着培养时间的延长ＩＣＡＭ１表达量逐渐增加。四氯化碳实验性大鼠肝纤维化模型中，新分离的ＨＳＣ中即可见ＩＣＡＭ１表达。结论体内动物实验和体外细胞培养表明，ＩＣＡＭ１表达可能与ＨＳＣ活化、肝脏炎性损伤及肝纤维化的发生有关。

Objective To explore the relationship between hepatic stellate cells (HSC) and expression of intercellular adhesion molecule1(ICAM1). Methods By in situ perfusion with proteinase and collagenase and density gradient centrifugation with Nycodenz, hepatic stellate cells (HSC) were isolated and cultured from liver of normal Wistar rats and that induced by carbon tetrachloride. Expression of ICAM1 in HSC was detected by immunohistochemistry assay and reverse transcriptionpolymerase chain reaction (RTPCR). Results Expression of ICAM1 was not found in freshly isolated HSC of normal rats, but was present in primary culture of HSC on day 10 and secondary culture of HSC on day 7, the intensify of expression increased with duration of culture. Expression of ICAM1 was observed at freshly isolated HSC in the liver of rats induced by carbon tetrachloride. Conclusions Expression of ICAM1 was related to activation of HSC, hepatic inflammation and liver fibrogenesis.