التفاصيل البيبلوغرافية
| العنوان: |
下调 miR-320a 表达对缺氧/复氧诱导的心肌细胞增殖和 凋亡的影响. (Chinese) |
| العنوان البديل: |
Effect of down-regulation of miR-320a expression on proliferation and apoptosis of cardiomyocytes induced by hypoxia/reoxygenation. (English) |
| المؤلفون: |
李红英, 王晨燕, 郭世超, 赵友为, 董彦博, 黄建成 |
| المصدر: |
Journal of Jilin University (Medicine Edition); Jul2023, Vol. 49 Issue 4, p958-967, 10p |
| مصطلحات موضوعية: |
GENE expression, BCL-2 proteins, JAK-STAT pathway, LACTATE dehydrogenase, REPORTER genes |
| الملخص (بالإنجليزية): |
Objective: To discuss the effect of down-regulation of miR-320a expression on the myocardial hypoxia/reoxygenation (H/R)injury model, and to clarify its related mechanism. Methods: Real-time fluorescence quantitative PCR (RT-qPCR)method was used to detect the expression levels of miR-320a in serum of the patients with actue myocardial infarction (AMI)and the myocardial H9C2 cells induced by H/R. The miR-320a inhibitor, inhibitor NC, small interference Janus kinase 2 (si-JAK2), and si-NC plasmids were transfected into the H9C2 cells respectively, and blank control group was set up. After successful transfection, the H/R treatment was performed. The H9C2 cells were divided into control group, H/R group, H/R + inhibitor NC group, H/R + miR-320a inhibitor group, H/R + miR-320a inhibitor + si-NC group and H/R + miR-320a inhibitor + si-JAK2 group. The targeting relationship between miR-320a and Janus kinase 2 (JAK2)was detected by double luciferase reporter gene; the proliferation rate of cells in various groups were detected by CCK-8 assay; the activities of superoxide dismutase (SOD)and levels of malonaldehyde (MDA)in cells and the levels of lactate dehydrogenase (LDH)in cell culture supernanant in various groups were detected by biochemical method; the apoptotic rates of cells in various groups were detected by flow cytometry; the expression levels of miR-320a and JAK2 mRNA in cells in various groups were detected by RT-qPCR method; the expression levels of B-cell lymphoma-2 (Bcl-2), Bcl-2 associated X protein (Bax), cleaved-cysteinyl aspartate specific proteinase-3 (cleaved-caspase-3), JAK2, signal transducers and activator of transcription 3 (STAT3), and phosphorylated STAT3 (p-STAT3)proteins in cells in various groups were detected by Western blotting method. Results: The expression levels of miR-320a in serum of the patients with AMI and the myocardial H9C2 cells in H/R group were significantly higher than those in control group (P<0. 05). The results of double Luciferase reporter gene detection suggested that miR-320a could targetedly bind with JAK2. Compared with control group, the proliferation rate of the cells and SOD activity in the cells in H/R group were decreased significantly (P<0. 05), the apoptotic rate of the cells, MDA level and LDH activity in the cells were significantly increased (P<0. 05), the expression levels of Bcl-2 and JAK2 proteins and ratio of p-STAT3/STAT3 in the cells were significantly decreased (P<0. 05), and the expression levels of Bax and cleaved caspase-3 proteins in the cells were significantly increased (P<0. 05). Compared with H/R group, the proliferation rate of the cells and SOD activity in the cells in H/R+miR-320a inhibitor group were increased (P<0. 05), while the apoptotic rate of the cells, MDA level in the cells, LDH activity in the cell culture supernanant were decreased (P<0. 05), the expression levels of Bcl-2 and JAK2 proteins and ratio of p-STAT3/STAT3 in the cells were significantly increased (P<0. 05), the expression levels of Bax and cleaved- caspase-3 proteins in the cells were significantly decreased (P<0. 05). Compared with H/R+miR-320a inhibitor+ si-NC group, the proliferation rate of the cells and SOD activity in the cells in H/R+miR-320a inhibitor+ si-JAK2 group were decreased (P<0. 05), and the apoptotic rate of the cells, MDA level in the cells, and LDH activity in the cell culture supernanant were increased (P<0. 05). Conclusion: Down-regulation of miR-320a expression can inhibit the apoptosis of the cardiomyocytes induced by H/R and increase the proliferation activity of cells, and its mechanism is related to the targeted regulation of JAK2/STAT3 signaling pathway. [ABSTRACT FROM AUTHOR] |
| Abstract (Chinese): |
目的：探讨下调 miR-320a 表达对心肌细胞缺氧/复氧 (H/R) 损伤模型的影响，并阐明其 相关作用机制。方法：采用实时荧光定量 PCR (RT-qPCR) 法检测急性心肌梗死 (AMI) 患者血清 中和 H/R 诱导的心肌 H9C2 细胞中 miR-320a 表达水平。将 miR-320a inhibitor、inhibitor NC、小干扰 Janus 激酶 (si-JAK2) 和 si-NC 质粒分别转染至 H9C2 细胞中，同时设空白对照组，确定转染成功后进 行 H/R 处理。H9C2 细胞分为对照组、H/R 组、H/R+inhibitor NC 组、H/R+miR-320a inhibitor 组、 H/R+miR-320a inhibitor+si-NC 组和 H/R+miR-320a inhibitor+si-JAK2 组。双荧光素酶报告基因检 测 miR-320a 与 Janus 激酶 2 (JAK2) 的靶向关系，CCK-8 法检测各组细胞增殖率，生化法检测各组细 胞中超氧化物歧化酶 (SOD) 活性、丙二醛 (MDA) 水平和细胞培养上清液中乳酸脱氢酶 (LDH) 活性，流式细胞术检测各组细胞凋亡率，RT-qPCR 法检测各组细胞中 miR-320a 和 JAK2 mRNA 表达 水平，Western blotting 法检测各组细胞中 B 淋巴细胞瘤 2 (Bcl-2)、Bcl-2 相关 X 蛋白 (Bax)、剪切型 含半胱氨酸的天冬氨酸蛋白水解酶 3 (cleaved-caspase-3)、JAK2、信号转导因子和转录激活因子 3 (STAT3) 及磷酸化 STAT3 (p-STAT3) 蛋白表达水平。结果：AMI 患者血清中和 H/R 组 H9C2 细 胞中 miR-320a 表达水平明显高于对照组 (P<0. 05)。双荧光素酶报告基因检测结果提示 miR-320a 可 与 JAK2 靶向结合。与对照组比较，H/R 组细胞增殖率和细胞中 SOD 活性明显降低 (P<0. 05)，细胞 凋亡率、细胞中 MDA 水平和细胞培养上清液中 LDH 活性明显升高 (P<0. 05)，细胞中 Bcl-2 和 JAK2 蛋白表达水平及 p-STAT3/STAT3 比值明显降低 (P<0. 05)，Bax 和 cleaved-caspase-3 蛋白表达水平 明显升高 (P<0. 05)。与 H/R 组比较，H/R+miR-320a inhibitor 组细胞增殖率和细胞中 SOD 活性明 显升高 (P<0. 05），细胞凋亡率、细胞中 MDA 水平和细胞培养上清液中 LDH 活性明显降低 (P< 0. 05)，细胞中 Bcl-2 和 JAK2 蛋白表达水平及 p-STAT3/STAT3 比值明显升高 (P<0. 05)，Bax 和 cleaved-caspase-3 蛋白表达水平明显降低 (P<0. 05)。与 H/R+miR-320a inhibitor+si-NC 组 比 较 ， H/R+miR-320a inhibitor+si-JAK2 组细胞增殖率和细胞中 SOD 活性明显降低 (P<0. 05)，细 胞 凋 亡 率、细胞中 MDA 水平和细胞培养上清液中 LDH 活性明显升高 (P<0. 05)。结论：下调 miR-320a 表 达可抑制 H/R 诱导的心肌细胞凋亡，增加细胞增殖活性，其作用机制与靶向调控 JAK2/STAT3 信号 通路有关。 [ABSTRACT FROM AUTHOR] |
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| قاعدة البيانات: |
Complementary Index |
