Department of Clinical Laboratory, The Second Affiliated Hospital of Harbin Medical University, Harbin, 150086, China.
In Vitro Cell Dev Biol Anim. 2024 Mar;60(2):151-160. doi: 10.1007/s11626-023-00842-7. Epub 2023 Dec 28.
Endothelial cell damage and dysfunction are crucial factors in the development and early stages of coronary artery disease (CAD) and apoptosis plays a significant role in this process. In this study, We aimed to simulate the CAD vascular microenvironment by treating endothelial cells with tumor necrosis factor alpha (TNF-α) to construct an endothelial cell apoptosis model. Our findings revealed that the TNF-α model resulted in increased micro-RNA 223-3p (miR-223-3p) mRNA and Bax protein expression, decreased kruppel-like factor 15 (KLF15) and Bcl-2 protein expression, and decreased cell viability. More importantly, in the TNF-α-induced endothelial cell apoptosis model, transfection with the miR-223-3p inhibitor reversed the effects of TNF-α on Bcl-2, Bax expression. We transfected miRNA-223-3p mimics or inhibitors into endothelial cells and assessed miR-223-3p levels using RT-PCR. Cell viability was detected using CCK8. Western blot technology was used to detect the expression of Bcl-2, Bax, and KLF15. In summary, this study demonstrates the role and possible mechanism of miR-223-3p in endothelial cells during CAD, suggesting that miR-223-3p may serve as a promising therapeutic target in CAD by regulating KLF15.
内皮细胞损伤和功能障碍是冠心病（CAD）发展和早期的关键因素，细胞凋亡在此过程中起着重要作用。本研究通过用肿瘤坏死因子-α（TNF-α）处理内皮细胞来模拟 CAD 血管微环境，构建内皮细胞凋亡模型。我们的研究结果表明，TNF-α 模型导致 micro-RNA 223-3p（miR-223-3p）mRNA 和 Bax 蛋白表达增加，Kruppel 样因子 15（KLF15）和 Bcl-2 蛋白表达降低，细胞活力降低。更重要的是，在 TNF-α诱导的内皮细胞凋亡模型中，转染 miR-223-3p 抑制剂可逆转 TNF-α对 Bcl-2、Bax 表达的影响。我们将 miRNA-223-3p 模拟物或抑制剂转染到内皮细胞中，并通过 RT-PCR 检测 miR-223-3p 水平。通过 CCK8 检测细胞活力。采用 Western blot 技术检测 Bcl-2、Bax 和 KLF15 的表达。总之，本研究表明 miR-223-3p 在 CAD 期间在内皮细胞中的作用和可能的机制，提示 miR-223-3p 可能通过调节 KLF15 成为 CAD 有前途的治疗靶点。
