Department of Cardiology, The First Peoples' Hospital of Jining, Jining, Shandong, China (mainland).
Department of Vascular Surgery, The First Peoples' Hospital of Jining, Jining, Shandong, China (mainland).
Med Sci Monit. 2019 Jul 30;25:5648-5656. doi: 10.12659/MSM.915263.
BACKGROUND Endothelial cells are involved in vascular homeostasis, and endothelial cell dysfunction is involved in the pathogenesis of cardiovascular disease. This study aimed to investigate the effects of microRNA-154 in human umbilical vein endothelial cells (HUVECs) following injury induced by hydrogen peroxide (H₂O₂). MATERIAL AND METHODS Cell viability and apoptosis of HUVECs treated with H₂O₂ were measured. The expression of microRNA-154 was detected by quantitative real-time polymerase chain reaction (qRT-PCR). Cell survival, caspase-3 activity, and the apoptosis rate were evaluated in H₂O₂-treated HUVECs cells after the upregulation and down-regulation of microRNA-154 expression. The interaction between microRNA-154 and Dickkopf WNT signaling pathway inhibitor 2 (DKK2) was predicted by bioinformatics analysis and was verified by luciferase reporter gene assay and Western blot. The effects of DKK2 short-interfering RNA (siRNA) on antioxidant injury in HUVECs cells were determined. RESULTS The survival rate of HUVECs exposed to H₂O₂ was significantly reduced and the apoptosis rate was significantly increased, and H₂O₂ significantly inhibited the expression of microRNA-154 in a dose-dependent manner. Overexpression of microRNA-154 increased cell survival, reduced the activity of caspase-3, and reduced cell apoptosis. Inhibition of microRNA-154 expression decreased cell survival, increased the activity of caspase-3, and promoted cell apoptosis. Luciferase reporter gene assay and Western blot showed that microRNA-154 interacted with the Wnt pathway molecule DKK2 in HUVECS. Also, DDK2 siRNA resulted in a similar protective effect on H₂O₂-treated HUVECs as overexpression of microRNA-154. CONCLUSIONS Oxidative injury in HUVECs was regulated by microRNA-154 targeting the Wnt/ß-catenin signaling pathway.
内皮细胞参与血管稳态，内皮细胞功能障碍与心血管疾病的发病机制有关。本研究旨在探讨过氧化氢（H₂O₂）诱导损伤后，微小 RNA-154 对人脐静脉内皮细胞（HUVEC）的影响。
用 H₂O₂ 处理 HUVEC 后，测量细胞活力和细胞凋亡。用实时定量聚合酶链反应（qRT-PCR）检测微小 RNA-154 的表达。上调和下调微小 RNA-154 表达后，评估 H₂O₂ 处理的 HUVEC 细胞中的细胞存活、半胱天冬酶-3 活性和细胞凋亡率。通过生物信息学分析预测微小 RNA-154 与 Dickkopf WNT 信号通路抑制剂 2（DKK2）之间的相互作用，并通过荧光素酶报告基因检测和 Western blot 进行验证。通过检测 HUVEC 细胞抗氧化损伤来确定 DKK2 短发夹 RNA（siRNA）的作用。
暴露于 H₂O₂ 的 HUVEC 细胞存活率显著降低，细胞凋亡率显著升高，且 H₂O₂ 呈剂量依赖性显著抑制微小 RNA-154 的表达。过表达微小 RNA-154 增加细胞存活率，降低半胱天冬酶-3 活性，减少细胞凋亡。抑制微小 RNA-154 表达降低细胞存活率，增加半胱天冬酶-3 活性，促进细胞凋亡。荧光素酶报告基因检测和 Western blot 显示，微小 RNA-154 与 HUVECS 中的 Wnt 通路分子 DKK2 相互作用。此外，DKK2 siRNA 对 H₂O₂ 处理的 HUVEC 产生了类似于过表达微小 RNA-154 的保护作用。
微小 RNA-154 通过靶向 Wnt/β-catenin 信号通路调节 HUVEC 的氧化损伤。
