Department of Cardiovascular Surgery, Daping Hospital, Army Medical University, Chongqing, China.
Eur Rev Med Pharmacol Sci. 2019 Apr;23(7):3058-3069. doi: 10.26355/eurrev_201904_17588.
In the clinic, therapeutic options for pulmonary arterial hypertension are limited; therefore, investigating the therapeutic strategies and novel therapies is critical for pulmonary arterial hypertension (PAH) treatment. This study aimed to evaluate the role of miRNA-126 (miR-126) and its associated signaling pathways and specific mechanisms for the pathogenesis of PAH.
The pulmonary artery endothelial cells (PAECs) were isolated and identified. The miR-126 mimic and miR-126 inhibitor were synthesized. LV-3-miR-126 mimic viral vector and LV-3-miR-126 inhibitor vector were established and infected into pulmonary artery endothelial cells. Expression of sprouty-related EVH1 domain-containing protein 1 (SPRED1), phosphoinositide-3-kinase regulatory subunit 2 (PIK3R2) and miR-126 were detected using Real-time PCR (RT-PCR). Cell apoptosis (Annexin V-PE/7-AAD) and proliferation (PKH26) were examined by using FACScan flow cytometry. Vascular endothelial growth factor (VEGF), transforming growth factor β1 (TGF-β1) and TGF-β3 levels were evaluated using enzyme-linked immunosorbent assay (ELISA) kits.
miR-126 inhibited the endothelial cells related to SPRED1 and PIK3R2 expression. Over-expression of miR-126 significantly inhibited the PAECs apoptosis compared to PAECs and blank LV-3 vector group (p<0.05). miR-126 significantly triggered the PAECs proliferation compared to PAECs and blank LV-3 vector group (p<0.05). In functional analysis, miR-126 mimic significantly increased the cells amounts of S phases compared to PAECs and blank LV-3 vector group (p<0.05). Pre-infection with miR-126 mimic significantly enhanced the levels of VEGF, TGF-β1, and TGF-β3 compared to PAECs and blank LV-3 vector group (p<0.05).
miR-126 could affect cell apoptosis, proliferation, cell cycle, and modulate VEGF/TGF-β levels.
在临床中，肺动脉高压的治疗选择有限；因此，研究治疗策略和新疗法对于肺动脉高压（PAH）的治疗至关重要。本研究旨在评估微小 RNA-126（miR-126）及其相关信号通路和特定机制在 PAH 发病机制中的作用。
分离和鉴定肺动脉内皮细胞（PAEC）。合成 miR-126 模拟物和 miR-126 抑制剂。建立 LV-3-miR-126 模拟病毒载体和 LV-3-miR-126 抑制剂载体，并感染肺动脉内皮细胞。采用实时 PCR（RT-PCR）检测芽生相关 EVH1 结构域包含蛋白 1（SPRED1）、磷酸肌醇 3-激酶调节亚基 2（PIK3R2）和 miR-126 的表达。采用 FACScan 流式细胞术检测细胞凋亡（Annexin V-PE/7-AAD）和增殖（PKH26）。采用酶联免疫吸附试验（ELISA）试剂盒检测血管内皮生长因子（VEGF）、转化生长因子β1（TGF-β1）和 TGF-β3 水平。
miR-126 抑制与 SPRED1 和 PIK3R2 表达相关的内皮细胞。与 PAEC 和空白 LV-3 载体组相比，过表达 miR-126 可显著抑制 PAEC 细胞凋亡（p<0.05）。与 PAEC 和空白 LV-3 载体组相比，miR-126 可显著触发 PAEC 细胞增殖（p<0.05）。在功能分析中，miR-126 模拟物可显著增加 S 期细胞数量，与 PAEC 和空白 LV-3 载体组相比（p<0.05）。与 PAEC 和空白 LV-3 载体组相比，miR-126 模拟物预感染可显著增加 VEGF、TGF-β1 和 TGF-β3 水平（p<0.05）。
miR-126 可影响细胞凋亡、增殖、细胞周期，并调节 VEGF/TGF-β 水平。
