干扰素-γ诱导大鼠肠黏膜微血管内皮细胞表达细胞黏附因子-1和干扰素-γ受体α

IFN-γ inducing ICAM-1 and IFN-γRα expression in cultured rat intestinal mucosa microvascular endothelial cells

目的以体外培养的大鼠肠黏膜微血管内皮细胞(RIMMVECs)为研究对象,探讨干扰素-γ(IFN-γ)对于体外培养的RIMMVECs表达细胞黏附因子-1(ICAM-1)和IFN-γ受体α(IFN-γRα)的影响。方法体外分离培养RIMMVECs,用不同浓度的IFN-γ对所培养的RIMMVECs进行诱导,通过荧光定量RT-PCR法和流式细胞术,从mRNA和蛋白水平检测活化后的RIMMVECs表达ICAM-1和IFN-γRα的情况。结果浓度为20μg/L和40μg/L的IFN-γ可以激活RIMMVECs,在mRNA和蛋白水平均能提高ICAM-1的表达,并且6 h时达到高峰;20μg/L的IFN-γ刺激RIMMVECs后,可使其上调表达IFN-γRαmRNA,但是蛋白表达量却随着刺激时间逐渐降低;而40μg/L的IFN-γ刺激RIMMVECs 2 h后,IFN-γRαmRNA达到峰值,6 h之后,其表达量呈下降趋势,IFN-γRα蛋白浓度也随之逐渐降低;10μg/L的IFN-γ对于ICAM-1和IFN-γRα的表达无影响。结论 IFN-γ可能是通过影响微血管内皮细胞(MVECs)表面IFN-γRα分子和ICAM-1的表达,从而调控和参与细胞免疫反应和炎症反应。

Objective To investigate the influences of interferon-γ （IFN-γ） on the intercellular cell adhesion molecule-1 （ICAM-1） and IFN-γRα expression in the cultured rat intestinal mucosa microvascular endothelial cells （RIMMVECs）. Methods After the RIMMVECs separation culture, IFN-γ in different doses were used to induce the in vitro cultured RIMMVECs, ICAM-1 and IFN-γRct expression in the cultured RIMMVECs at both the mRNA and protein levels which were detected by real-time PCR and flow cytometry, respectively. Results The results showed that RIMMVECs were activated by IFN-γ/ at the concentrations of 20μg/L and 40 μg/L. ICAM-1 expression increased in the activated RIMMVECs at both mRNA and protein levels, reaching peak effects at 6 hours; IFN-γ/Rα mRNA expression was increased, but the protein decreased after stimulation by 20 p,g/L; IFN-γ/Rct mRNA expression peaked at 2 hours and then decreased after 6 hours when stimulated by 40 μg/L IFN-γ and the expression of IFN--γRα protein decreased gradually; 10 μg/L IFN-γhad no effect on the expressions of ICAM-1 and IFN-γ/Rα at both gene and protein levels. Conclusion These results suggest that IFN-γ may function to modulate and participate in the cellular immune response and inflammatory responses by influencing IFN-γ/Rα and ICAM-1 expression on intestinal mucosa microvascular cells.