Department of Inspection, Jiaozhou Central Hospital Of Qingdao, Shandong Province, Qingdao, P.R. China.
Eur Rev Med Pharmacol Sci. 2020 May;24(9):5008-5017. doi: 10.26355/eurrev_202005_21192.
To investigate the effects and mechanism of miR-145-5p on hypoxia/reoxygenation (H/R)-induced cardiac microvascular endothelial cell (CMEC) injury in coronary heart disease (CHD).
Patients with CHD (n=96) and healthy volunteers (n=96) were enrolled, and H/R injury model of CMECs was established. The expression of miR-145-5p and mothers against decapentaplegic homolog 4 (Smad4) mRNA in cells was quantified with reverse transcription polymerase chain reaction (RT-PCR). Then, miR-145-5p mimics and Smad4 inhibitor were transfected into CMECs. Cell counting kit-8 (CCK-8) was employed for proliferation detection, flow cytometry for apoptosis detection, and Western Blot for measuring the expression of apoptosis-related proteins and Smad4 protein.
The expression of serum miR-145-5p in patients with CHD was significantly lower than that in healthy individuals. The area under the curve (AUC) of miR-145-5p in diagnosing CHD was 0.894, and the expression of miR-145-5p was negatively correlated with that of Smad4 (p<0.05). Over-expression of miR-145-5p promoted the proliferation, inhibited the apoptosis, and reduced inflammatory responses and oxidative stress in H/R-injured CMECs. Moreover, miR-145-5p might negatively regulate the expression of Smad4 in CMECs. Dual-Luciferase reporter assay determined the targeting relation between miR-145-5p and Smad4.
MiR-145-5p is lowly expressed in patients with CHD, and its over-expression effectively alleviates H/R-induced CMEC injury by inhibiting Smad4.
探讨 miR-145-5p 对冠心病（CHD）患者缺氧/复氧（H/R）诱导的心肌微血管内皮细胞（CMEC）损伤的作用及机制。
选取 CHD 患者（n=96）和健康志愿者（n=96），建立 CMECs H/R 损伤模型。采用逆转录聚合酶链反应（RT-PCR）定量检测细胞中 miR-145-5p 和母亲对抗 decapentaplegic 同源物 4（Smad4）mRNA 的表达。然后，将 miR-145-5p 模拟物和 Smad4 抑制剂转染到 CMECs 中。采用细胞计数试剂盒-8（CCK-8）检测细胞增殖，流式细胞术检测细胞凋亡，Western blot 检测凋亡相关蛋白和 Smad4 蛋白的表达。
CHD 患者血清 miR-145-5p 表达明显低于健康人群。miR-145-5p 诊断 CHD 的曲线下面积（AUC）为 0.894，miR-145-5p 表达与 Smad4 呈负相关（p<0.05）。过表达 miR-145-5p 可促进 H/R 损伤的 CMEC 增殖，抑制其凋亡，并减轻炎症反应和氧化应激。此外，miR-145-5p 可能负调控 CMECs 中 Smad4 的表达。双荧光素酶报告基因实验确定了 miR-145-5p 与 Smad4 之间的靶向关系。
CHD 患者 miR-145-5p 表达降低，过表达 miR-145-5p 通过抑制 Smad4 有效减轻 H/R 诱导的 CMEC 损伤。
