Linc-NeD125通过调控微小RNA-19b对缺氧/复氧诱导的心肌细胞增殖和凋亡的影响  

Effects of Linc-NeD125 on hypoxia/reoxygenation-induced cardiomyocyte proliferation and apoptosis by regulating miR-19b expression

作　　者：薛晓燕 曹文婷 付媖 Xue Xiaoyan;Cao Wenting;Fu Ying(Department of Cardiology,Shanxi Bethune Hospital,Shanxi Academy of Medical Sciences,Tongji Shanxi Hospital,Third Hospital of Shanxi Medical University,Taiyuan,Shanxi 030032,China)

机构地区：[1]山西白求恩医院(山西医学科学院同济山西医院)山西医科大学第三医院心内科,山西太原030032

出　　处：《中国药物与临床》2023年第8期515-519,I0002,共6页Chinese Remedies & Clinics

摘　　要：目的 探究上调长链非编码RNA(lncRNA)Linc-NeD125对缺氧/复氧心肌细胞增殖和凋亡的影响及相关分子机制。方法 采用缺氧/复氧诱导HCM细胞建立损伤模型,通过实时定量聚合酶链反应(RT-q PCR)技术检测缺氧/复氧诱导的HCM细胞Linc-NeD125表达水平。将阴性对照慢病毒和Linc-NeD125慢病毒分别感染缺氧/复氧诱导的HCM细胞,HCM细胞分为NC组和Linc-NeD125组。细胞计数(CCK-8)法和流式细胞术分别检测NC组和Linc-NeD125组HCM细胞的增殖水平和凋亡率。双荧光素酶报告基因实验检测Linc-NeD125与微小RNA(miR)-19b的靶向关系。RT-qPCR检测NC组和Linc-NeD125组HCM细胞中miR-19b表达水平。蛋白免疫印迹法检测NC组和Linc-NeD125组HCM细胞中细胞周期蛋白(Cdk2、Cyclin E)、抗凋亡蛋白(Bcl-XL、Mcl-1)以及促凋亡蛋白Bax的表达。结果 缺氧/复氧诱导HCM细胞中LincNeD125表达水平低于正常HCM细胞(P<0.01)。Linc-NeD125组HCM细胞中Linc-NeD125表达水平高于NC组(P<0.01)。与NC组相比,Linc-NeD125组HCM细胞的增殖水平升高(P<0.05),细胞凋亡率降低(P<0.01)。双荧光素酶报告基因实验证实Linc-NeD125可与miR-19b靶向结合(P<0.01)。与NC组相比,Linc-NeD125组HCM细胞中miR-19b表达水平降低(P<0.01)。与NC组相比,Linc-NeD125组HCM细胞中细胞周期蛋白Cdk2、Cyclin E表达水平均升高(P<0.01),抗凋亡蛋白Bcl-XL、Mcl-1表达水平均升高(P<0.01),促凋亡蛋白Bax表达水平降低(P<0.01)。结论 上调Linc-NeD125能够增加缺氧/复氧诱导的心肌细胞增殖活性,抑制心肌细胞的凋亡,其分子机制与靶向调控miR-19b表达有关。Objective To explore the effects of up-regulation of long non-coding RNA(lncRNA)Linc-NeD125 on the proliferation and apoptosis of hypoxic/reoxygenated cardiomyocytes and the related molecular mechanisms.Methods Hypoxia/reoxygenation-induced HCM cells were used to establish an injury model,and real-time quantitative polymerase chain reaction(RT-qPCR)technology was used to detect the expression level of Linc-NeD125 in hypoxia/reoxygenation-induced HCM cells.Negative control lentivirus and Linc-NeD125 lentivirus were infected with hypoxia/reoxygenation-induced HCM cells respectively,and HCM cells were divided into NC group and Linc-NeD125 group.CCK-8 method and flow cytometry were used to detect the proliferation level and apoptosis rate of HCM cells in the NC group and Linc-NeD125 group respectively.Dual-luciferase reporter gene experiment was used to detect the targeting relationship between Linc-NeD125 and miR-19b.RT-qPCR was used to detect the expression levels of miR-19b in HCM cells of NC group and Linc-NeD125 group.Western blot method was used to detect the expression of cell cycle proteins(Cdk2,Cyclin E),anti-apoptotic proteins(Bcl-XL,Mcl-1)and pro-apoptotic protein Bax in HCM cells of NC group and Linc-NeD125 group.Results The expres-sion level of Linc-NeD125 in hypoxia/reoxygenation-induced HCM cells was significantly lower than that in normal HCM cells(P<0.01).The expression level of Linc-NeD125 in HCM cells in the Linc-NeD125 group was signifi-cantly higher than that in the NC group(P<0.01).Compared with NC group,the proliferation level of HCM cells in the Linc-NeD125 group was significantly increased(P<0.05),and the cell apoptosis rate was significantly decreased(P<0.01).Dual-luciferase reporter gene experiment confirmed that Linc-NeD125 could target and bind to miR-19b(P<0.01).Compared with NC group,the expression level of miR-19b in HCM cells in the Linc-NeD125 group was significantly reduced(P<0.01).Compared with NC group,the expression levels of cell cycle proteins Cdk2 and Cyclin E in HCM cells of

关 键 词：RNA 长链非编码 肌细胞 心脏 微RNAS 

分 类 号：R542.22[医药卫生—心血管疾病]