机构地区:[1]青岛市市立医院本部心内一科,山东青岛266011 [2]寿光市中医医院心血管病科,山东寿光262700 [3]青岛市市立医院东院区干部保健科,山东青岛266071
出 处:《实用检验医师杂志》2022年第1期78-81,共4页Chinese Journal of Clinical Pathologist
基 金:山东省青岛市公共领域科技支撑计划项目(2012-1-3-1-3)。
摘 要:目的研究微小RNA(miR)-21在缺氧环境中人脐静脉内皮细胞(HUVEC)凋亡中的作用。方法取原代HUVEC随机分为6组,包括正常对照组(在正常培养环境中培养,不进行任何处理)、缺氧处理6 h组、12 h组、24 h组(将细胞置于厌氧盒中进行缺氧培养6 h、12 h、24 h)、转染miR-21抑制剂+缺氧24 h组(转染miR-21抑制剂24 h后,给予缺氧处理24 h)、阴性转染+缺氧24 h组(转染miR-21抑制剂阴性对照24 h后,给予缺氧处理24 h)。处理完毕后收集各组细胞,采用实时荧光定量聚合酶链反应(RT-PCR)检测正常对照组和单纯缺氧各组细胞中miR-21的表达量;采用流式细胞术检测正常对照组和各处理组细胞的凋亡率;比较各组上述指标的差异。结果①缺氧处理6 h、12 h、24 h组miR-21的表达量均明显低于正常对照组〔缺氧6 h:0.75±0.07比1,缺氧12 h:0.71±0.07比1,缺氧24 h:0.66±0.06比1,均P<0.05〕。②缺氧处理6 h、12 h、24 h组的细胞凋亡率均明显高于正常对照组〔缺氧6 h(%):8.80±1.69比3.02±0.40,缺氧12 h(%):9.84±1.55比3.02±0.40,缺氧24 h(%):11.50±2.00比3.02±0.40,均P<0.05〕。转染miR-21抑制剂+缺氧24 h组的细胞凋亡率明显高于单纯缺氧24 h组(%:16.10±1.34比11.50±2.00,P<0.05);阴性转染+缺氧24 h组的凋亡率与单纯缺氧24 h组比较差异无统计学意义。结论HUVEC在缺氧刺激后miR-21表达水平降低,且缺氧时间不同,miR-21表达量不同;转染miR-21抑制剂后细胞凋亡率明显升高。因此,miR-21在细胞缺氧刺激过程中可能发挥抗细胞凋亡的作用。Objective To investigate the role of microRNA(miR)-21 in the apoptosis of hypoxic human umbilical vein endothelial cells(HUVEC).Methods The primary HUVEC were replaced and randomly divided into 6 groups,including normal control group(cultured in normal environment without any treatment),hypoxia stimulation for 6 hours,12 hours and 24 hours groups(the cells were placed in an anaerobic box for hypoxia culture for 6 hours,12 hours and 24 hours),transfection with miR-21 inhibitor+hypoxia for 24 hours group(24 hours after transfection with miR-21 inhibitor,hypoxia treatment for 24 hours)and negative transfection+hypoxia for 24 hours group(24 hours after transfection with miR-21 inhibitor negative control,hypoxia treatment for 24 hours).After stimulation,the cells in each group were collected,and the expression of miR-21 in normal control and hypoxia groups was detected by real-time fluorescence quantitative polymerase chain reaction(RT-PCR);the apoptosis rate of cells in each group was detected by flow cytometry.The differences of the above indexes in each group were compared.Results①The expression levels of miR-21 in hypoxia for 6 hours,12 hours and 24 hours groups were lower than that in normal control group[hypoxia for 6 hours:0.75±0.07 vs.1,hypoxia for 12 hours:0.71±0.07 vs.1,hypoxia for 24 hours:0.66±0.06 vs.1,all P<0.05].②The cell apoptosis rates in hypoxia for 6 hours,12 hours and 24 hours groups were higher than that in normal control group[hypoxia for 6 hours(%):8.80±1.69 vs.3.02±0.40,hypoxia for 12 hours(%):9.84±1.55 vs.3.02±0.40,hypoxia for 24 hours(%):11.50±2.00 vs.3.02±0.40,all P<0.05].The apoptosis rate of transfection with miR-21 inhibitor+hypoxia for 24 hours group was higher than that of hypoxia for 24 hours group(%:16.10±1.34 vs.11.50±2.00,P<0.05);there was no significant difference in apoptosis rate between negative transfection+hypoxia for 24 hours group and hypoxia for 24 hours group.Conclusions After hypoxia stimulation,the expression of miR-21 in HUVEC decreased,and the amount was
关 键 词:微小RNA-21 缺氧 人脐静脉内皮细胞 细胞凋亡
分 类 号:R329.2[医药卫生—人体解剖和组织胚胎学]
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