缺氧对人肺动脉平滑肌细胞内活性氧及细胞色素C的影响及意义  

The effect and significance of hypoxia on ROS and cytochrome C in hPASMCs

作　　者：陈乾 沈乐[1,2] 杨静 张建银[1,2] 耑冰[1,2] 赵霞[1,2] 李丹丹 董辉[3] CHEN Qian;SHEN Le;YANG Jin;ZHANG Jianyin;ZHUAN Bing;ZHAO Xia;LI Dandan;DONG Hui(Department of Respiratory Medicine,People's Hospital of Ningxia Hui Autonomous Region,Yinchuan 750002,China;Department of Respiratory Medicine,the First Affiliated Hospital of Northwest Minzu University,Yinchuan 750002,China;Scientific Research Center,General Hospital of Ningxia Medical University,Yinchuan 750004,China)

机构地区：[1]宁夏回族自治区人民医院呼吸内科,宁夏银川750002 [2]西北民族大学第一附属医院呼吸内科,宁夏银川750002 [3]宁夏医科大学总医院科研中心,宁夏银川750004

出　　处：《宁夏医学杂志》2022年第1期55-58,F0003,共5页Ningxia Medical Journal

基　　金：国家自然科学基金项目(82160017);宁夏重点研发项目(2018BEG03054、2019BEG03045);宁夏回族自治区人民医院培育振兴项目(201709)。

摘　　要：目的观察缺氧对人肺动脉平滑肌细胞(hPASMCs)内活性氧(ROS)及细胞色素C表达的影响,探索缺氧性肺动脉高压的形成机制。方法购买hPASMCs,分别在常氧条件和缺氧条件下进行培养,连续细胞培养72h,并观察细胞增殖情况;使用CCK-8法分别于细胞培养后的1、24、48及72 h连续观察细胞增殖情况,使用分子探针对上述时间节点hPASMCs内的ROS水平进行检测,使用Westernblot检测hPASMCs内细胞色素C的表达水平。结果常氧组与缺氧组在1、24、48及72 h的培养过程中,其细胞增殖水平的OD值分别为缺氧组(0.306±0.004、0.733±0.009、0.973±0.018和1.258±0.030)、常氧组(0.307±0.004、0.459±0.022、0.751±0.021和1.151±0.037),2组细胞的增殖水平差异有统计学意义(P<0.05),缺氧组的hPASMCs增殖水平明显高于常氧组;2组hPASMCs在1、24、48及72 h的培养过程中,其细胞内ROS荧光OD值差异分别为缺氧组(2.32±0.192、3.46±0.270、4.12±0.259和4.7±0.187)、常氧组(2.38±0.130、2.30±0.158、2.34±0.207和2.38±0.179),2组hPASMCs内ROS荧光OD值比较差异有统计学意义(P<0.05),缺氧组的hPASMCs内ROS水平明显高于常氧组;2组hPASMCs在1、24、48及72 h的培养过程中,其细胞内细胞色素C的表达相对值分别为缺氧组(0.516±0.029、0.673±0.060、0.981±0.135和1.180±0.131)、常氧组(0.509±0.064、0.488±0.023、0.534±0.016和0.512±0.033),2组hPASMCs内细胞色素C的表达相对值差异有显著性(P<0.05),缺氧组hPASMCs内细胞色素表达水平明显高于常氧组。结论缺氧将导致hPASMCs明显增殖,其内ROS水平增加,同时伴有细胞色素C水平增加,这种改变可能参与了肺动脉血管重构,并最终诱发了肺动脉高压的形成。Objective To observe the effect of hypoxia on the expression of ROS and cytochrome C in hPASMCs and explore the mechanism of hypoxic pulmonary hypertension.Methods Purchasing hPASMCs and culturing them under normoxic and hypoxic conditions for 72 hours to observe the cell proliferation.CCK-8 method was used to continuously observe cell proliferation at 1h,24h,48h and 72 h after cell culturing.In this case,molecular probes were used to detect the ROS level in hPASMCs at the above time points,and western blot was used to detect the expression level of cytochrome C in hPASMCs.Results The OD values of the cell proliferation level were 0.306±0.004,0.733±0.009,0.973±0.018,1.258±0.030 at the 1 h,24 h,48 h and 72 h in the hypoxic group,respectively,and the OD values were 0.307±0.0039,0.459±0.022,0.751±0.021 and 1.151±0.037 in the normal group,the differences in the proliferation levels between two groups were statistically significant(P<0.05).The proliferation level of hPASMCs in the hypoxia group was significantly higher than that in the normal group.The ROS fluorescence OD values of hPASMCs were 2.32±0.1923,3.46±0.270,4.12±0.259 and 4.7±0.187 at 1 h,24 h,48 h and 72 h points in the hypoxia group,respectively,and the ROS fluorescence OD values were 2.38±0.130,2.30±0.158,2.34±0.207 and 2.38±0.179 in the normoxia group,the difference of the ROS fluorescence OD values between two groups was statistically significant(P<0.05),and the ROS levels in the hPASMCs in the hypoxia group were significantly higher than in the normoxic group.The relative values of cytochrome C were 0.516±0.029,0.673±0.060,0.981±0.135 and 1.180±0.131 at 1 h,24 h,48 h and 72 h points in the hypoxia group,respectively,and the relative values of cytochrome C were 0.509±0.064,0.488±0.023,0.534±0.016 and 0.512±0.033 in the normoxia group,and the difference between them was statistically significant(P<0.05),the expression level of cytochrome C in hPASMCs in the hypoxia group was significantly higher than that of the normoxic group.Con

关 键 词：缺氧 人肺动脉平滑肌细胞 活性氧 细胞色素C 

分 类 号：R563[医药卫生—呼吸系统]