肺动脉平滑肌细胞对低氧诱导的肺微血管内皮细胞白介素表达的调节及机制  

Regulatory effects and mechanisms of pulmonary arterial smooth muscle cells on the hypoxia-induced interleukins expression in pulmonary microvascular endothelial cells

作　　者：张志远[1] 钱桂生[1] 王建春[1] 李运成[1] 刘婷[1] 王关嵩[1] 

机构地区：[1]第三军医大学新桥医院呼吸疾病研究所,重庆400037

出　　处：《解放军医学杂志》2009年第6期725-728,共4页Medical Journal of Chinese People's Liberation Army

基　　金：国家自然科学基金资助项目(30770928);军队"十一五"重点攻关课题资助项目(06G083;08G093);国家教育部留学回国人员科研启动基金项目(2007)

摘　　要：目的研究低氧对肺微血管内皮细胞(PMVECs)的白细胞介素(IL)6、IL-1β、受体酪氨酸激酶(RTK)和Janus激酶3(JAK3)表达的影响,以及肺动脉平滑肌细胞(PASMCs)在其中的调控作用。方法单独培养的大鼠PMVECs分为正常(N)组及低氧2h(H2)组、6h(H6)组、12h(H12)组,另设立PMVECs共培养组(与PASMCs共同培养)和单纯培养组,均接受6h或12h的低氧(氧浓度为3%)处理。应用RT-PCR方法检测IL-6、JAK3的mRNA表达水平,放免法测定培养细胞上清中的IL-6蛋白含量,放射性酶分析法测定细胞膜、胞质中RTK的活性,ELISA法测定细胞培养上清中的IL-1β蛋白含量。结果低氧刺激后,PMVECs的IL-6、JAK3mRNA表达及IL-6、IL-1β蛋白含量均增加。与N组比较,低氧处理各组的以上各指标均有所上升(P<0.01),以6h时间点最为明显(P<0.01)。胞膜RTK活性在低氧刺激各组明显降低(P<0.01),而胞质RTK活性明显升高(P<0.01),以12h时间点最为明显(P<0.05)。在低氧刺激6h后,共培养组的PMVECs内IL-6、JAK3的mRNA表达,以及IL-6、IL-1β蛋白含量均较单纯培养组有所降低(P<0.05);在低氧刺激12h后,共培养组的PMVECs的RTK活性也较单纯培养组降低(P<0.05)。结论低氧可以促进PM-VECs的IL-6mRNA表达,并能增加IL-6和IL-1β蛋白含量,复合培养情况下PASMCs对此具有下调作用,其机制可能与RTK和JAK涉及的信号转导通路有关。Objective To study the effects of hypoxia on the expression of interleukin-6 （IL 6）, interleukin-1β （IL-1β）, receptor tyrosine kinase （RTK） and Janus kinase 3 （JAK3） in pulmonary microvascular endothelial cells （PMVECs）, and the regulatory effects of the co-cultured pulmonary arterial smooth muscle cells （PASMCs）. Methods PMVECs were divided into 4 groups： normal group （N）, 2h hypoxia group （H2）, 6h hypoxia group （H6） and 12h hypoxia group （H12）. Moreover, PMVECs, co-cultured with PMVECs or solitarily cultured, were divided into co-culture group and solitary group, respectively, and they were exposed to 3% concentration of oxygen for 6h or 12h. The mRNA expressions of IL-6 and JAK3 were detected by RT-PCR, the levels of IL-6 and IL-1β in the supematant were measured by radioimmunoassay and ELISA respectively, and the activities of RTK on the membrane and cytoplasm were detected by radioactive enzyme assay. Results Compared with that in N group, the mRNA expressions of IL-6 and JAK3, as well as the protein concentrations of IL-6 and IL-1β increased after hypoxia treatment （P〈0. 01）, and peaked at 6h time point （P〈0. 01）. The activity of RTK on membrane was significantly decreased （P〈0. 01）, but increased in cytoplasm （P〈0. 01） in the hypoxia-treated groups in comparison with that in N group, and the maximal effects occurred at the 12h time point （P〈0. 05）. The mRNA expressions of IL-6 and JAK3, and the concentrations of IL-6 and IL-1β in co-cultured group decreased significantly （P〈0. 05） compared with that in solitary group （P〈0. 05） after being treated by hypoxia for 6 hours. The RTK activity of co-cultured group was lower than that of solitary group after hypoxia treatment for 12 hours （P〈0. 05）. Conclusion Hypoxia may facilitate the mRNA expression of IL-6, as well as the protein expressions of IL-6 and IL-1β in PMVECs, which can be down-regulated by PASMCs via RTK and JAK3 signaling pathways.

关 键 词：肺动脉 肌细胞 平滑肌 内皮细胞 细胞低氧 白细胞介素 受体蛋白质酪氨酸激酶类 Janus激酶3 

分 类 号：R364.4[医药卫生—病理学]