| 苏 婷,宋晓伟,史承勇,唐文栋,胡海鹰,郑 兴.PRKAG2-AS1通过AMPK抑制缺氧所致心肌细胞凋亡[J].,2020,(4):608-613 |
| PRKAG2-AS1通过AMPK抑制缺氧所致心肌细胞凋亡 |
| PRKAG2-AS1 is Involved in the Regulation of Apoptosis in Myocardial Hypoxia Injury |
| 投稿时间：2019-09-30 修订日期：2019-10-26 |
| DOI：10.13241/j.cnki.pmb.2020.04.002 |
| 中文关键词: PRKAG2-AS1 LncRNA 心肌细胞凋亡 |
| 英文关键词: PRKAG2-AS1 LncRNA Cardiomyocyte apoptosis |
| 基金项目:国家自然科学基金项目(81170092) |
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| 中文摘要: |
| 摘要 目的：探讨PRKAG2-AS1在缺氧所致心肌细胞凋亡中的作用及其可能机制。方法：以人心肌细胞系作为主要研究对象，使用RNA核质分提的方法检测PRKAG2-AS1在细胞中的表达分布模式。将人心肌细胞系分为常氧对照组及低氧组，分别置于常氧环境(21 % O2)和低氧环境(1 % O2)培养12小时，构建心肌细胞缺氧模型，AnnexinV-FITC/PI流式细胞学检测细胞凋亡，Real-time PCR检测模型中SOD mRNA及PRKAG2-AS1基因表达。对常氧培养条件下心肌细胞通过siRNA及反寡义核苷酸方法分别靶向敲低胞质及胞核内PRKAG2-AS1的表达水平，AnnexinV-FITC/PI流式细胞学检测细胞凋亡，观察PRKAG2-AS1对心肌细胞凋亡的影响；Real-time PCR检测SOD mRNA表达，Western blot检测AMPKγ2亚基蛋白的表达，观察PRKAG2-AS1对SOD mRNA及AMPKγ2蛋白表达的影响。结果：PRKAG2-AS1在心肌细胞胞质及胞核中均有表达，且以胞核为主。PRKAG2-AS1基因表达水平在心肌细胞缺氧模型中明显降低(P＜0.05)。对常氧培养条件下心肌细胞，敲低PRKAG2-AS1基因表达，将导致细胞凋亡增加(P＜0.05)，且敲低胞核中表达细胞凋亡更为明显，同时，敲低PRKAG2-AS1能够引起SOD mRNA表达水平改变(P＜0.05)，且AMPKγ2蛋白表达水平降低(P＜0.05)。结论：PRKAG2-AS1可能通过AMPK途径影响SOD表达，从而调控心肌缺氧损伤中的细胞凋亡。 |
| 英文摘要: |
| ABSTRACT Objective: This study was aimed to explore the effect and possible mechanism of PRKAG2-AS1 in the apoptosis in myocardial hypoxia injury. Methods: The human cardiomyocyte cell line (AC16) was selected as the main research object, RNA nuclear fractionation was taken to find out the expression pattern of PRKAG2-AS1 in the cell. AC16 cells were divided into control group and hypoxia group, treated under hypoxia (1% O2) or normoxia (21% O2) condition for 12 hours. AnnexinV-FITC/PI flow cytometry was used to detect the apoptosis rate, realtime-PCR was performed to detect PRKAG2-AS1 RNA expression level. Furthermore, PRKAG2-AS1 siRNA and anti-oligonucleotide was used to investigate the role of PRKAG2-AS1 on apoptosis of cardiomyocytes under normoxia condition, the mRNA levels of SOD and protein expression of AMPK were detected by realtime-PCR and Western-blot respectively. Results: The results shown that PRKAG2-AS1 was expressed in the cytoplasm and nucleus, mainly in the nucleus (P<0.05). Compared with the control group, the RNA relative expression level of PRKAG2-AS1 was significantly decreased in the hypoxic model of cardiomyocytes(P<0.05). PRKAG2-AS1 siRNA and oligo could promote apoptosis in AC16 cells under normoxia condition (P<0.05), which was more obvious by oligo. Moreover, the mRNA expression levels of SOD changed(P<0.05), and the protein level of AMPKγ2 were reduced (P<0.05). Conclusion: PRKAG2-AS1 may regulate apoptosis in myocardial hypoxia injury by regulating the expression of SOD through AMPK pathway. |
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