摘要
目的 建立肺缺血再灌注细胞模型并研究BCL-2结合抗凋亡基因(BCL-2 associated anthanogen-1,Bag-1)在肺缺血再灌注中的表达,以期为研究Bag-1在肺缺血再灌注疾病的作用机制提供实验基础。方法 本课题以A549细胞系作为肺缺血再灌注模型的细胞模型,实验分5组,将A549细胞给予不同缺氧时间:0 h、6 h、12 h、18 h、24 h缺氧,再复氧24 h处理后,通过低温、低氧、葡萄糖剥夺建立缺血再灌注细胞模型。通过比较5组细胞活性、乳酸脱氢酶(lactate dehydrogenase,LDH)以及活性氧自由基(reactive oxygen species,ROS)水平,确定建模效果,同时观察Bag在肺缺血再灌注中的表达。结果 不同缺氧时间处理后,缺氧组细胞活性均较正常组细胞活性降低(P<0.01),并随缺氧时间延长细胞活性下降,各时间点间细胞活性存在显著差异(F=85.03,P<0.001)。缺氧组LDH、ROS浓度均较正常组细胞明显升高(P<0.01),各时间点间LDH(F=107.67,P<0.001)、ROS(F=42.61,P<0.001)水平具有统计学意义,Bag-1表达在缺氧6 h时最高,后随时间延长水平逐渐下降。结论 以A549细胞系作为模型细胞,以低温、低氧、葡萄糖剥夺为方法可成功建立缺血再灌注细胞模型,Bag-1在缺血再灌注损伤中表达趋势为先升高,再降低。
Objective To establish a lung ischemia-reperfusion cell model and study the expression of BCL-2 associated anthangen-1(Bag-1) in lung ischemia-reperfusion,in order to provide an experimental basis for studying the mechanism of action of Bag-1 in lung ischemia-reperfusion diseases.Methods In this study,A549 cell line was used as the cell model of lung ischemia-reperfusion model,and the experiment was divided into 5 groups.A549 cells were given different hypoxia time:0 h,6 h,12 h,18 h,24 h,and then reoxygenated for 24 h,and the ischemia-reperfusion cell model was established by hypothermia,hypoxia and glucose deprivation.Cell activity,lactate dehydrogenase(LDH) and reactive oxygen species(ROS) levels of the five groups were compared to determine the modeling effect,and the expression of Bag during lung ischemia reperfusion was observed.Results The cell activity of the hypoxia group was lower than that of the normal group after different hypoxia duration treatment(P<0.01),and cell activity decreased with the extension of hypoxia time,and there were significant differences in cell activity at each time point(F=85.03,P<0.001).The concentrations of LDH and ROS in the hypoxia group were significantly higher than those in the normal group [At each time point,LDH( F = 107.67,P < 0.001),ROS( F = 42.61,P <0.001) ],and there was statistical significance.Bag-1 expression was the highest at 6H of hypoxia,and then decreased with the extension of time.Conclusion With A549 cell line as the model cell,hypothermia,hypoxia and glucose deprivation as the methods,the ischemia-reperfusion cell model can be successfully established.The expression of Bag-1 in ischemia-reperfusion injury tends to increase at first and then decrease.
作者
王成
彭兴男
李泽明
王越
李洋
吕纪玲
WANG Cheng;PENG Xingnan;LI Zeming;WANG Yue;LI Yang;L Jiling(Department of Respiratory and Critical Care Medicine,the Second People’s Hospital of Shandong Province,Jinan,Shandong 250022,China)
出处
《临床肺科杂志》
2024年第4期541-545,共5页
Journal of Clinical Pulmonary Medicine
基金
山东省老年医学学会科技发展计划项目(No.LKJGG2021W099)。
