沉默LncRNA SNHG7通过调控miR-181b-5p的表达减轻缺氧/复氧诱导的心肌细胞损伤  

Silencing LncRNA SNHG7 alleviates hypoxia/reoxygenation-induced cardiomyocyte damage by regulating the expression of miR-181b-5p

作　　者：刘振[1] 金卫东[1] 韩明磊[1] 崔佳佳 侯永兰[1] 徐光翠[2] Liu Zhen;Jin Weidong;Han Minglei;Cui Jiajia;Hou Yonglan;Xu Guangcui(Department of Cardiology,Xinxiang Central Hospital,Xinxiang,Henan 453000,China;Xinxiang Medical College,Xinxiang,Henan 453000,China)

机构地区：[1]新乡市中心医院心血管内科,453000 [2]新乡医学院,453000

出　　处：《中华医学遗传学杂志》2021年第8期812-817,共6页Chinese Journal of Medical Genetics

基　　金：国家自然科学基金(81703183)。

摘　　要：目的探讨沉默长链非编码RNA SNHG7(LncRNA SNHG7)对缺氧/复氧(hypoxia/reoxygenation,H/R)诱导的心肌细胞损伤的影响及其对miR-181b-5p的靶向调控作用。方法体外培养大鼠H9c2心肌细胞,将其随机分为对照组、H/R组、H/R+si-NC组、H/R+si-SNHG7组、H/R+si-SNHG7+anti-miR-NC组和H/R+si-SNHG7+anti-miR-181b-5p组。检测乳酸脱氢酶、丙二醛的含量与超氧化物歧化酶的活性;用流式细胞术检测细胞凋亡率;用实时荧光定量PCR(qRT-PCR)检测SNHG7、miR-181b-5p的表达量;用双荧光素酶报告实验验证SNHG7、miR-181b-5p的靶向关系;蛋白免疫印迹法检测Bax和Bcl-2蛋白的表达量。结果与对照组相比,H/R组心肌细胞中SNHG7的表达显著升高(P<0.05),miR-181b-5p的表达水平显著降低(P<0.05),乳酸脱氢酶、丙二醛的含量显著升高(P<0.05),超氧化物歧化酶的活性显著降低(P<0.05),细胞凋亡率显著升高(P<0.05),Bax蛋白水平显著升高(P<0.05),Bcl-2蛋白水平显著降低(P<0.05);与H/R组、H/R+si-NC组相比,H/R+si-SNHG7组乳酸脱氢酶、丙二醛的含量显著降低(P<0.05),超氧化物歧化酶的活性显著升高(P<0.05),细胞凋亡率显著降低(P<0.05),Bax蛋白水平显著降低(P<0.05),Bcl-2蛋白水平显著升高(P<0.05);双荧光素酶报告实验证实SNHG7可靶向结合miR-181b-5p;干扰miR-181b-5p的表达可减弱沉默SNHG7对H/R诱导的心肌细胞氧化应激及细胞凋亡的作用。结论沉默SNHG7可能通过上调miR-181b-5p的表达抑制H/R诱导的心肌细胞氧化应激及细胞凋亡,从而对心肌细胞发挥保护作用。Objective To study the effect of silencing LncRNA SNHG7 on hypoxia/reoxygenation(H/R)-induced cardiomyocyte injury and its targeted regulation on miR-181b-5p.Methods Rat cardiomyocytes H9c2 were cultured in vitro and randomly divided into control group,H/R group,H/R+si-NC group,H/R+si-SNHG7 group,H/R+si-SNHG7+anti-miR-NC group and H/R+si-SNHG7+anti-miR-181b-5p group.The content of lactate dehydrogenase(LDH),malondialedhyde(MDA)and the activity of superoxide dismutase(SOD)were detected.Flow cytometry was carried out to detect the rate of apoptosis.qRT-PCR was used to detect the expression of SNHG7 and miR-181b-5p.Dual luciferase report experiment was used to verify the targeting relationship between SNHG7 and miR-181b-5p.Western blotting was used to detect the expression of Bax and Bcl-2.Results Compared with the control group,the H/R group showed significantly increased SNHG7 expression in cardiomyocytes,reduced miR-181b-5p expression,higher levels of LDH and MDA,reduced activity of SOD,increased cell apoptosis rate,higher level of Bax protein,and reduced level of Bcl-2 protein(all P<0.05).Compared with the H/R and H/R+si-NC groups,the H/R+si-SNHG7 group had significantly reduced level of LDH and MDA,increased activity of SOD,reduced apoptosis rate,reduced level of Bax protein,increased level of Bcl-2 protein(all P<0.05).The dual luciferase report experiment confirmed that SNHG7 could target miR-181b-5p.Interference with the expression of miR-181b-5p could reduce the effect of silencing SNHG7 on H/R-induced cardiomyocyte oxidative stress and apoptosis.Conclusion Silencing SNHG7 may inhibit H/R-induced cardiomyocyte oxidative stress and apoptosis by up-regulating the expression of miR-181b-5p,thereby exerting a protective effect on cardiomyocytes.

关 键 词：长链非编码RNA SNHG7 miR-181b-5p 缺氧/复氧 心肌细胞 氧化应激 凋亡 

分 类 号：R542.22[医药卫生—心血管疾病]