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lncRNASOX21-AS1靶向miR-590-3p对支气管上皮细胞缺氧/复氧损伤的影响
作者:王江燕1  秦晓娟2  满小佳1 
单位:1. 成都市龙泉驿区妇幼保健院 内科, 四川 成都 610100;
2. 四川省医学科学院·四川省人民医院 全科医学中心, 四川 成都 610100
关键词:长链非编码RNA SOX21反义RNA1 miR-590-3p 支气管上皮细胞 氧化应激 损伤 
分类号:R562.2
出版年·卷·期(页码):2022·50·第三期(342-348)
摘要:

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-2B 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.

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