Department of Cardiology, Weanpon Industry 521 Hospital, China.
Department of Cardiology, Affiliated Hospital of Yan'an University, China.
Kaohsiung J Med Sci. 2021 Apr;37(4):314-323. doi: 10.1002/kjm2.12333. Epub 2020 Dec 18.
The current study aims to explore the miRNA changes that occur in the serum of patients with coronary heart disease (CHD) and healthy controls using a microarray technique, thereby exploring the potential biomarkers in the diagnosis of CHD and the underlying mechanism. Clinical data were reviewed, and venous blood samples were collected from 66 cases of CHD and 58 cases of healthy controls. MicroRNA-wide expression profiling identified 16 miRNAs that were aberrantly decreased by ~2-fold in the serum of patients with CHD compared to that of healthy controls. RT-PCR analysis indicated that the expression of miR-3129-5p was increased the most in patients with CHD compared with that of controls. Moreover, serum miR-3129-5p was found to be highest in the severe stenosis group, followed by the moderate stenosis group and mild stenosis group. ROC analysis showed that serum miR-3129-5p could differentiate patients with CHD from controls. Further study showed that mTOR was a target gene of miR-3129-5p. Western blot assays demonstrated that miR-3129-5p significantly suppressed the phosphorylation of S6 but increased LC3II/LC3I and Beclin1 levels. Consistently, GFP-LC3 and TEM assays indicated that miR-3129 increased autophagy puncta in H9C2 cells. More importantly, silencing mTOR significantly decreased the expression of p-S6 but increased LC3II/LC3I and Beclin expression even in H9C2 cells transfected with miR-3129-5p inhibitor, indicating that miR-3129-5p-induced cell autophagy was mediated via mTOR in H9C2 cells. In summary, elevated serum miR-3129-5p contributes to CHD by targeting mTOR signaling and may be a therapeutic target in the treatment of CHD.
本研究旨在利用微阵列技术探讨冠心病（CHD）患者与健康对照者血清中 miRNA 的变化，从而探索 CHD 诊断中的潜在生物标志物及相关机制。回顾临床资料，采集 66 例 CHD 患者和 58 例健康对照者的静脉血标本。miRNA 全表达谱分析鉴定出 16 个 miRNA 在 CHD 患者血清中较健康对照者降低约 2 倍。RT-PCR 分析表明，与对照组相比，CHD 患者 miR-3129-5p 的表达增加最多。此外，血清 miR-3129-5p 在重度狭窄组最高，中度狭窄组次之，轻度狭窄组最低。ROC 分析显示血清 miR-3129-5p 可区分 CHD 患者与对照组。进一步研究表明 mTOR 是 miR-3129-5p 的靶基因。Western blot 检测表明 miR-3129-5p 可显著抑制 S6 的磷酸化，但增加 LC3II/LC3I 和 Beclin1 水平。同样，GFP-LC3 和 TEM 检测表明 miR-3129 可增加 H9C2 细胞中的自噬斑点。更重要的是，沉默 mTOR 可显著降低 p-S6 的表达，增加 LC3II/LC3I 和 Beclin 的表达，即使在转染 miR-3129-5p 抑制剂的 H9C2 细胞中也是如此，表明 miR-3129-5p 诱导的细胞自噬是通过 H9C2 细胞中的 mTOR 介导的。总之，血清 miR-3129-5p 升高通过靶向 mTOR 信号通路促进 CHD 的发生，可能是 CHD 治疗的一个潜在靶点。
