Department of Neurosurgery, The Third People's Hospital of Henan Province, Zhengzhou 450006, China.
Department of Neurosurgery, The First Affiliated Hospital of Zhengzhou University, Zhengzhou, 450052, China.
Biosci Rep. 2024 May 29;44(5). doi: 10.1042/BSR20231511.
Vascular endothelial cell premature senescence plays an important part in stroke. Many microRNAs (miRNAs) are known to be involved in the pathological process of vascular endothelial cell premature senescence. The present study aimed to investigate the mechanism of hydrogen peroxide (H2O2)-induced premature senescence in human umbilical vein endothelial cells (HUVECs) and effect of miR-142-3p on hydrogen peroxide (H2O2)-induced premature senescence. HUVECs were exposed to H2O2 to establish a model premature senescence in endothelial cells. CCK-8 assay was performed to detect cell viability. Senescence-associated β-galactosidase staining assay and senescence-related proteins p16 and p21 were used to detect changes in the degree of cell senescence. RT-qPCR and Western blot were conducted to measure mRNA and protein levels, respectively. The scratch wound-healing assay, transwell assay, and EdU assay were performed to evaluate the ability of migration and proliferation, respectively. miRNA-142-3p and silencing information regulator 2 related enzyme 1 (SIRT1) binding was verified using Targetscan software and a dual-luciferase assay. We found that miRNA-142-3p is abnormally up-regulated in HUVECs treated with H2O2. Functionally, miRNA-142-3p inhibition may mitigate the degree of HUVEC senescence and improve HUVEC migration and proliferation. Mechanistically, SIRT1 was validated to be targeted by miRNA-142-3p in HUVECs. Moreover, SIRT1 inhibition reversed the effects of miRNA-142-3p inhibition on senescent HUVECs exposed to H2O2. To our knowledge, this is the first study to show that miRNA-142-3p ameliorates H2O2-induced HUVECs premature senescence by targeting SIRT1 and may shed light on the role of the miR-142-3p/SIRT1 axis in stroke treatment.
血管内皮细胞衰老在中风中起着重要作用。已知许多 microRNAs（miRNAs）参与了血管内皮细胞衰老的病理过程。本研究旨在探讨过氧化氢（H2O2）诱导人脐静脉内皮细胞（HUVEC）衰老的机制及 miR-142-3p 对 H2O2 诱导的内皮细胞衰老的影响。用 H2O2 处理 HUVEC 以建立内皮细胞衰老模型。用 CCK-8 法检测细胞活力。衰老相关β-半乳糖苷酶染色法和衰老相关蛋白 p16 和 p21 用于检测细胞衰老程度的变化。用 RT-qPCR 和 Western blot 分别检测 mRNA 和蛋白水平。划痕愈合实验、Transwell 实验和 EdU 实验分别用于评估迁移和增殖能力。用 Targetscan 软件和双荧光素酶实验验证 miRNA-142-3p 与沉默信息调节因子 2 相关酶 1（SIRT1）的结合。我们发现 H2O2 处理的 HUVEC 中 miRNA-142-3p 异常上调。功能上，miRNA-142-3p 抑制可减轻 HUVEC 衰老程度，并改善 HUVEC 迁移和增殖。机制上，在 HUVEC 中验证了 SIRT1 是 miRNA-142-3p 的靶基因。此外，SIRT1 抑制逆转了 miRNA-142-3p 抑制对 H2O2 处理的衰老 HUVEC 的作用。据我们所知，这是第一项表明 miRNA-142-3p 通过靶向 SIRT1 改善 H2O2 诱导的 HUVEC 衰老的研究，可能为 miR-142-3p/SIRT1 轴在中风治疗中的作用提供启示。
