Department of Cardiology, The First Affiliated Hospital of Zhengzhou University, Zhengzhou, Henan, China.
Eur Rev Med Pharmacol Sci. 2017 Dec;21(23):5456-5461. doi: 10.26355/eurrev_201712_13935.
To identify the expression changes of microRNA 93 (miR-93) in oxygen-glucose deprivation/reoxygenation (OGD/R) injury in cardiomyocytes and its mechanism of mediating OGD/R and inducing apoptosis.
Primary cardiomyocytes were extracted and OGD/R model in cardiomyocytes was established in vitro. Quantitative reverse transcription polymerase chain reaction (qRT-PCR) was used to detect the expressions of miR-93, and Western blot assay was applied to measure the protein levels of nuclear factor erythroid 2-related factor 2 (Nrf2) and caspase-3. Flow cytometry was utilized to examine the cardiomyocyte apoptosis rate.
The apoptosis rate was increased after OGD/R in cardiomyocytes, accompanied by remarkable rise of miR-93 expression. After transfection of miR-93 antagomir, the apoptosis rate of cardiomyocyte induced by OGD/R was down-regulated, and the expression of cleaved caspase-3 was decreased. Meanwhile, the results of qRT-PCR and Western blot showed that the levels of Nrf2 mRNA and protein expression were up-regulated after the miR-93 level was inhibited, and luciferase reporter assay affirmed that Nrf2 was a target molecule for OGD/R-induced apoptosis mediated by miR-93.
miR-93 mediates OGD/R-induced hypoxia/reoxygenation injury apoptosis in cells by targeting Nrf2.
鉴定微小 RNA 93（miR-93）在心肌细胞氧葡萄糖剥夺/复氧（OGD/R）损伤中的表达变化及其介导 OGD/R 诱导细胞凋亡的机制。
体外提取原代心肌细胞，建立心肌细胞 OGD/R 模型。采用实时定量逆转录聚合酶链反应（qRT-PCR）检测 miR-93 的表达，Western blot 法检测核因子红细胞 2 相关因子 2（Nrf2）和半胱氨酸天冬氨酸蛋白酶-3（caspase-3）的蛋白水平。流式细胞术检测心肌细胞凋亡率。
OGD/R 后心肌细胞凋亡率增加，miR-93 表达明显升高。转染 miR-93 拮抗剂后，OGD/R 诱导的心肌细胞凋亡率下调，caspase-3 裂解表达减少。同时，qRT-PCR 和 Western blot 结果显示，miR-93 水平抑制后 Nrf2 mRNA 和蛋白表达水平上调，荧光素酶报告实验证实 Nrf2 是 miR-93 介导的 OGD/R 诱导细胞凋亡的靶分子。
miR-93 通过靶向 Nrf2 介导 OGD/R 诱导的细胞缺氧/复氧损伤凋亡。
