外源性一氧化碳对大鼠肺动脉平滑肌细胞凋亡状况的影响  

Effects of low-dose carbon monoxide on expression of caspase-3 in rat pulmonary artery smooth muscle cells

作　　者：李运成[1] 贺斌峰[2] 钱桂生[2] 王关嵩[2] 

机构地区：[1]蚌埠市第一人民医院呼吸科,233000 [2]第三军医大学新桥医院全军呼吸内科研究所,重庆400037

出　　处：《中华肺部疾病杂志（电子版）》2016年第5期494-497,共4页Chinese Journal of Lung Diseases(Electronic Edition)

基　　金：国家自然科学基金资助项目(81370168);蚌埠市科技计划项目[(2004)10]

摘　　要：目的研究外源性一氧化碳对大鼠肺动脉平滑肌细胞(pulmonary artery smooth muscle cells,PASMC)凋亡状况的影响,探讨低氧肺血管重建的发病机制及防治措施。方法复合胶原酶法培养动脉平滑肌细胞,分别给予常氧、低氧、低氧合并低浓度一氧化碳处理。采用透射电镜、流式细胞仪、逆转录聚合酶链式反应(reverse transcription-polymerase chain reaction,RT-PCR)、免疫组织化学染色等方法检测肺动脉平滑肌细胞的凋亡状况及caspase-3表达水平。结果电镜观察可见,低浓度一氧化碳处理PASMC后呈凋亡样改变,染色质浓缩成块状位于核周边,细胞核边集近似半月型,细胞器结构完整。流式细胞仪示,正常培养PASMC没有明显的亚二倍峰,低氧24 h条件下可见亚二倍峰,低浓度一氧化碳处理24 h后,PASMC亚二倍峰明显升高。RT-PCR和免疫组织化可见,低浓度一氧化碳作用后caspase-3的mRNA和蛋白水平显著升高。结论低浓度一氧化碳可以调节caspase-3基因的表达,从而促进肺动平滑肌细胞的凋亡。Objective To investigate the influence of low-dose carbon monoxide on the expression of caspase-3 in rat pulmonary artery smooth muscle cells（PASMC） , and explore the pathogenesis and prenvention measures of pulmonary vasecular remoding. Methods The PASMCs of rats were cultured in normal O2, hypoxia, combined hypoxia and carbon monoxide condition. Eleetronmicroseope, flow cytometry, reverse transcription-polymerase chain reaction （RT-PCR） and immunohistochemistrycal methods were used to detect the expression of the caspase-3 in rat pulmonary artery smooth muscle cells. Result In the PASMCs of rats cultured in combined hypoxia and carbon monoxide condition, the typical morphological changes of apoptosis were observed under transmission electronmicroscope, such as chromatin condensation and aggregation at the nuclear periphery, and nuclear fragmentation as well. The flow eytometer showed that typical apoptosis peak appeared in combined hypoxia and carbon monoxide condition, however, it was not obvious in hypoxia and normal O2 condition. Caspase-3 mRNA and protein expression were significantly increased in combined hypoxia and carbon monoxide condition compared with those in hypoxia and normal 02 condition. Conclusions Low concentration of carbon monoxide can regulate the expression of caspase-3, thereby promoting apoptosis of pulmonary artery smooth muscle cells.

关 键 词：低氧性肺动脉高压 一氧化碳 平滑肌细胞 CASPASE-3 

分 类 号：R322.12[医药卫生—人体解剖和组织胚胎学] R329-3[医药卫生—基础医学]