lncRNA SOX21- AS1靶向miR-590-3p对支气管上皮细胞缺氧/复氧损伤的影响

Effect of lncRNA SOX21-AS1 targeting miR-590-3p on hypoxia/reoxygenation injury bronchial epithelial cells

目的:探讨长链非编码RNA(lncRNA)SOX21反义RNA1(SOX21-AS1)对支气管上皮细胞缺氧/复氧损伤的影响及分子机制。方法:将人支气管上皮细胞系BEAS-2B分为Con组、缺氧/复氧组(H/R组)、缺氧/复氧+si-NC组(H/R+si-NC组)、缺氧/复氧+si-SOX21-AS1组(H/R+si-SOX21-AS1组)、缺氧/复氧+miR-NC组(H/R+miR-NC组)、缺氧/复氧+miR-590-3p组(H/R+miR-590-3p组)、缺氧/复氧+si-SOX21-AS1+anti-miR-NC组(H/R+si-SOX21-AS1+anti-miR-NC组)、缺氧/复氧+si-SOX21-AS1+anti-miR-590-3p组(H/R+si-SOX21-AS1+anti-miR-590-3p组)。实时荧光定量PCR(RT-qPCR)检测SOX21-AS1和miR-590-3p的表达水平;试剂盒分别检测培养液中乳酸脱氢酶(LDH)水平和细胞中超氧化物歧化酶(SOD)活性及丙二醛(MDA)含量;流式细胞术检测细胞凋亡;双荧光素酶报告实验检测SOX21-AS1和miR-590-3p的靶向关系。结果:缺氧/复氧诱导的支气管上皮细胞中SOX21-AS1表达水平升高,miR-590-3p表达水平降低(P<0.001),LDH的水平升高,SOD的活性降低,MDA的含量升高,细胞凋亡率升高(P<0.05)。抑制lncRNA SOX21-AS1表达或过表达miR-590-3p,支气管上皮细胞中LDH水平降低,SOD活性升高,MDA含量降低,细胞凋亡率降低(P<0.05)。SOX21-AS1靶向调控miR-590-3p,干扰miR-590-3p表达逆转了抑制lncRNA SOX21-AS1表达对支气管上皮细胞缺氧/复氧损伤的作用。结论:抑制lncRNA SOX21-AS1表达可能通过上调miR-590-3p抑制缺氧/复氧诱导的支气管上皮细胞损伤。

Objective:To investigate the effect and molecular mechanism of long noncoding RNA(lncRNA)SOX21 antisense RNA1(SOX21-AS1)on hypoxia/reoxygenation injury bronchial epithelial cells.Methods:The human bronchial epithelial cell line BEAS-2 B was divided into Con group,hypoxia/reoxygenation group(H/R group),H/R+si-NC group,H/R+si-SOX21-AS1 group,H/R+miR-NC group,H/R+miR-590-3p group,H/R+si-SOX21-AS1+anti-miR-NC group and H/R+si-SOX21-AS1+anti-miR-590-3p group.Real-time fluorescence quantitative PCR(RT-qPCR)was used to detect the expressions of SOX21-AS1 and miR-590-3p;the kits were applied to assay the levels of lactate dehydrogenase(LDH)in the culture medium,the activity of superoxide dismutase(SOD)and malondialdehyde(MDA)content in the cells;flow cytometry to monitor apoptosis;the dual luciferase reporter experiment was performed to detect the targeting relationship between SOX21-AS1 and miR-590-3p.Results:In hypoxic/reoxygenated bronchial epithelial cells,the expression of SOX21-AS1 was increased,the expression of miR-590-3p was decreased(P<0.001),the level of LDH was increased,the activity of SOD was decreased,the content of MDA was increased,and the cells apoptosis rate was increased(P<0.05).Inhibited the expression of lncRNA SOX21-AS1 or overexpressed miR-590-3p,LDH level was decreased,SOD activity was increased,MDA content was decreased,apoptosis rate was decreased(P<0.05).SOX21-AS1 targets and regulates miR-590-3p,and interfering with miR-590-3p expression reverses the effect of inhibiting lncRNA SOX21-AS1 expression on hypoxia/reoxygenation injury of bronchial epithelial cells.Conclusion:Inhibition of lncRNA SOX21-AS1 expression may inhibit hypoxia/reoxygenation-induced bronchial epithelial cell injury by up regulating miR-590-3p.