Yue J-N, Li W-M, Hong W-Z, Yang J, Zhu T, Fang Y, Fu W-G
Department of Vascular Surgery, Zhongshan Hospital, Fudan University, Shanghai, China.
Eur Rev Med Pharmacol Sci. 2019 Aug;23(3 Suppl):319-326. doi: 10.26355/eurrev_201908_18663.
To investigate the effect of micro ribonucleic acid (miR)-210 on the apoptosis of vascular endothelial cells in arteriosclerosis obliterans (ASO) through the Janus kinase-signal transducer and activator of transcription (JAK-STAT) signaling pathway.
In the present work, the vascular endothelial cells in ASO patients were selected as objects of study, the cell lines with miR-210 interference and overexpression were constructed with the Crisp/Case9 technique for subsequent experiments as experimental group, and the aortic endothelial cells of a healthy human were used as control group. First, the changes in the transcriptional and translational levels of such genes as JAK2 and STAT3 in the JAK-STAT signaling pathway in cell lines with miR-210 interference and overexpression were detected via fluorescence quantitative Polymerase Chain Reaction (qPCR) and Western blotting. The changes in the transcriptional and translational levels of nitric oxide synthase (NOS) in cells were detected in experimental group and control group to clarify the regulatory effect of miR-210 on the JAK-STAT signaling pathway. At the same time, the cell proliferation in experimental group and control group was observed via methyl thiazolyl tetrazolium (MTT) assay and the apoptosis rate was detected in both groups via flow cytometry.
The results of fluorescence qPCR and Western blotting revealed that the expression level of miR-210 was significantly increased in cells of ASO patients compared with that in aortic endothelial cells of healthy human with a significant difference (p<0.05). At the same time, the inhibition on miR-210 could significantly reduce the transcriptional and translational levels of JAK2, STAT3, and NOS, block the JAK-STAT signaling pathway, suppress the cell proliferation, and promote apoptosis. The overexpression of miR-210 could markedly increase the transcriptional and translational levels of JAK2, STAT3, and NOS, activate the JAK-STAT signaling pathway, promote the cell proliferation, and suppress the apoptosis.
MiR-210 can be involved in the apoptosis process of vascular endothelial cells in ASO through the JAK-STAT signaling pathway.
通过Janus激酶-信号转导子和转录激活子（JAK-STAT）信号通路，研究微小核糖核酸（miR）-210对动脉硬化闭塞症（ASO）血管内皮细胞凋亡的影响。
本研究选取ASO患者的血管内皮细胞作为研究对象，采用Crisp/Case9技术构建miR-210干扰及过表达细胞系用于后续实验作为实验组，选取健康人主动脉内皮细胞作为对照组。首先，通过荧光定量聚合酶链反应（qPCR）和蛋白质免疫印迹法检测miR-210干扰及过表达细胞系中JAK-STAT信号通路相关基因JAK2和STAT3的转录及翻译水平变化。检测实验组和对照组细胞中一氧化氮合酶（NOS）的转录及翻译水平变化，以阐明miR-210对JAK-STAT信号通路的调控作用。同时，通过噻唑蓝（MTT）法观察实验组和对照组细胞增殖情况，并采用流式细胞术检测两组细胞凋亡率。
荧光定量PCR和蛋白质免疫印迹法结果显示，与健康人主动脉内皮细胞相比，ASO患者细胞中miR-210表达水平显著升高，差异有统计学意义（p<0.05）。同时，抑制miR-210可显著降低JAK2、STAT3和NOS的转录及翻译水平，阻断JAK-STAT信号通路，抑制细胞增殖并促进凋亡。miR-210过表达可显著提高JAK2、STAT3和NOS的转录及翻译水平，激活JAK-STAT信号通路，促进细胞增殖并抑制凋亡。
MiR-210可通过JAK-STAT信号通路参与ASO血管内皮细胞凋亡过程。
