沉默长链非编码RNA NR_027324对低糖缺氧损伤大鼠H9C2心肌细胞凋亡的影响  

Silencing of LncRNA NR_027324 Promotes Apoptosis Caused by Hypoglycemic and Hypoxic Injury in H9C2 Cells

作　　者：孟庆坤[1] 谭昊[2] 顾卉[3] 骆佳莹 王菁菁[3] 王传合[3] 韩苏[3] 孙志军[3] MENG Qing-Kun;TAN Hao;GU Hui;LUO Jia-Ying;WANG Jing-Jing;WANG Chuan-He;HAN Su;SUN Zhi-Jun(Internal Medicine-Cardiovascular Department,the Peoples Hospital of Liaoning Province(The People's Hospital of China Medical University),Shenyang 110000,China;Department of digestive and urinary soft radiotherapy,Liaoning Cancer Hospital(Cancer Hospital of China Medical University),Shenyang 110042,China;Intermal Medicine-Cardiovascvlar Department Shengjing Hospital of China Medical University,Shenyang 110000,China)

机构地区：[1]辽宁省人民医院(中国医科大学人民医院)心内科,沈阳110067 [2]辽宁省肿瘤医院(中国医科大学肿瘤医院)消化泌尿骨软放疗科,沈阳110042 [3]中国医科大学附属盛京医院心内科,沈阳110020

出　　处：《中国药物经济学》2022年第2期102-106,共5页China Journal of Pharmaceutical Economics

摘　　要：目的探讨在低糖缺氧大鼠H9C2细胞中,沉默长链非编码RNA(lncRNA)NR_027324的表达对H9C2细胞凋亡的影响。方法将H9C2心肌细胞在低糖缺氧环境中培养以构建心肌缺血缺氧模型。采用转染小干扰RNA(small interfering RNA,si-RNA)技术沉默NR_027324。利用逆转录定量聚合酶链反应(qRT-PCR)法测定lncRNA NR_027324的表达变化。检测乳酸脱氢酶(LDH)活性评估细胞损伤,MTT法检测细胞成活率,蛋白质免疫印迹法检测凋亡相关蛋白的相应变化。结果与对照组相比,低糖缺氧大鼠H9C2细胞中,NR_027324表达量明显上调,LDH升高,细胞成活率下降(P<0.001)。沉默NR_027324的表达后,低糖缺氧大鼠H9C2细胞中NR_027324表达量明显下调,LDH进一步升高,MTT法检测提示细胞成活率进一步下降(P<0.001)。低糖缺氧大鼠H9C2细胞中,Bax,cleaved-caspase3和cleaved-caspase9表达增加,Bcl-2表达下降(P<0.001)。沉默NR_027324的表达后,低糖缺氧大鼠H9C2细胞中Bax,cleaved-caspase3和cleaved-caspase9表达进一步增加,Bcl-2表达进一步下降(P<0.001)。结论低糖缺氧大鼠H9C2细胞中,沉默NR_027324的表达能够促进低糖缺氧H9C2细胞的凋亡,可能作为研究调控缺血缺氧心肌细胞凋亡的新靶点。Objective To investigate the silencing effect of lncRNA NR;27324 on apoptosis caused by hypoglycemic and hypoxic injury in H9C2 cells of rats.Methods The myocardial ischemia model in H9C2 cardiomyocytes was established by culture in hypoglycemia and hypoxia environment(OGD,oxygen-glucose deprivation).Small interfering RNA(siRNA)transfection was used to silence NR;27324.Reverse transcription-quantitative PCR(qRT-PCR)was performed to detect the expression levels of lncRNA NR;27324 mRNA.Cell injury and survival were assessed by lactate dehydrogenase(LDH)release and MTT,The expression of apoptosis-related proteins was detected by western blot.Results Compared with the control group,the expression levels of NR;27324 were significantly upregulated,LDH was elevated,the cell survival was decreased which was detected by MTT assay in the hypoglycemic and hypoxic H9C2 cells(P<0.001).After silencing NR;27324 in hypoglycemic and hypoxic H9C2 cells,the expression levels of NR;27324 were significantly down-regulated,LDH release was increased,the cell survival was decreased which was detected by MTT assay(P<0.001).The protein level of pro-apoptosis protein(Bax,cleaved-caspase3 and cleaved-caspase9)was elevated,while the anti-apoptotic protein Bcl-2 was reduced in hypoglycemic and hypoxic H9C2 cells(P<0.001).After silencing NR;27324 in hypoglycemic and hypoxic H9c2 cells,the pro-apoptosis proteins were elevated further and anti-apoptotic protein showed opposite changes(P<0.001).Conclusion Silencing NR;27324 can promote the apoptosis in the hypoglycemic and hypoxic H9C2 cells,which may be considered as a novel target to study the regulation of apoptosis of myocardial cells under ischemia and hypoxia.

关 键 词：低糖缺氧损伤 长链非编码RNA NR_027324 沉默RNA 转染小干扰RNA技术 凋亡 

分 类 号：R541.4[医药卫生—心血管疾病]