| 杜虹,张志远,谷小卫,等.微小 RNA-138-5p通过调控 Kelch重复蛋白影响缺氧 /复氧心肌细胞增殖与凋亡[J].安徽医药,2022,26(3):582-586. |
| 微小 RNA-138-5p通过调控 Kelch重复蛋白影响缺氧 /复氧心肌细胞增殖与凋亡 |
| Effect of miR-138-5p on proliferation and apoptosis of hypoxia/reoxygenated cardiomyocytes by regulating KLHDC10 |
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| DOI：10.3969/j.issn.1009-6469.2022.03.038 |
| 中文关键词: 心肌再灌注损伤 微小 RNA-138-5p Kelch重复蛋白 缺氧 /复氧 H9C2 细胞存活 细胞凋亡 |
| 英文关键词: Myocardial reperfusion injury MiR-138-5p KLHDC10 H/R H9C2 Cell survival Apoptosis |
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| 中文摘要: |
| 目的探讨微小 RNA(miRNA/miR)-138-5p、Kelch重复蛋白( KLHDC10)在缺氧 /复氧( H/R)心肌细胞 H9C2中增殖、凋亡的功能及二者的作用关系。方法 2018年 5月至 2019年 12月，建立 H/R H9C2细胞模型；正常培养的 H9C2细胞记为对照组。用 miRNA阴性对照( miR-con)、 miR-138-5p模拟物( miR-138-5p mimics)、过表达空载体( pcDNA-con)和 KLHDC10过表达载体( pcDNA-KLHDC10)不同处理方法处理 H/R H9C2细胞。实时荧光定量逆转录聚合酶链反应( qRT-PCR)、蛋白质印迹法(Western blotting)检测细胞中 miR-138-5p、KLHDC10、细胞周期蛋白 D1(cyclin D1)、活化胱天蛋白酶 -3(cleaved-caspase-3)的表达水平；细胞计数试剂盒( CCK-8)法和流式细胞术检测细胞的存活和凋亡；双荧光素酶报告基因实验检测细胞的荧光素酶活性。结果 H/R组于对照组， H/R H9C2细胞中 miR-138-5p的表达水平明显降低［( 0.34±0.03)比( 1.00±0.11)］KLHDC10的表达水平明显升高，细胞存活率显著降低［( 62.03±6.20)%比( 100.04±10.05)%］，凋亡率显著升高［( 28.16±2.8，2)%比( 7.22± |
| 英文摘要: |
| Objective To investigate the functions of microRNA (miRNA/miR)-138-5p and Kelch repeat protein (KLHDC10) in theproliferation and apoptosis of hypoxia/reoxygenation (H/R) cardiomyocytes (H9C2) and their relationship.Methods From May 2018 toDecember 2019, H/R H9C2 cell model was established and conventionally cultured H9C2 cells were used as the control group (NCgroup).H/R H9C2 cells were treated with different treatment methods: miRNA negative control (miR-con), miR-138-5p mimics (miR138-5p mimics), over-expressed empty vector (pcDNA-con) and KLHDC10 over-expressed vector (pcDNA-KLHDC10). Real-time fluorescence quantitative reverse transcriptase-polymerase chain reaction (qRT-PCR) and Western blotting were used to detect miR-1385p,KLHDC10, cyclin D1 and Lysed cysteine aspartic proteinase 3 (cleaved-caspase-3) expression levels; cell counting kit (CCK-8) andflow cytometry were used to detect cell survival and apoptosis. The double luciferase reporter assay was used to detect the luciferase activity of the cells.Results The H/R group was Compared with the NC group, the expression level of miR-138-5p was significantly reduced [(0.34±0.03) vs. (1.00±0.11)], the expression level of KLHDC10 was significantly increased, the cell survival rate was significantly reduced [(62.03±6.20)% vs. (100.04±10.05)%], and the apoptosis rate was significantly increased [ (28.16±2.82)% vs. (7.22±0.72)%] in H/R H9C2 cells. Meanwhile, the protein level of cyclin D1 was down-regulated and that of cleaved-caspase-3 was up-regulated (P< 0.05). Over-expressed miR-138-5p or inhibited KLHDC10 could significantly promote the survival and inhibit the apoptosis, up-regulate cyclin D1, and down-regulate cleaved-caspase-3 protein levels of H/R H9C2 cells. miR-138-5p significantly inhibited the luciferase activity of H9C2 cells in wild-type KLHDC10, and negatively regulated the KLHDC10 protein level in H9C2 cells; overexpressedKLHDC10 could reverse the regulatory role miR-138-5p in the survival and apoptosis of H/R H9C2 cells.Conclusion miR-138-5p could promote the survival and inhibit the apoptosis of H/R H9C2 cells, and one of the mechanisms is to targetedly down-regulate KLH DC10. |
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