Li W-M, Yue J-N, Guo D-Q, Fu W-G
Department of Vascular Surgery, Zhongshan Hospital, Fudan University, Shanghai, China.
Eur Rev Med Pharmacol Sci. 2019 Aug;23(3 Suppl):327-333. doi: 10.26355/eurrev_201908_18664.
To investigate the influence of micro ribonucleic acid (miR)-126 on the rats with lower limb arteriosclerosis obliterans (ASO).
Male Sprague- Dawley rats aged 3 months old were randomly divided into Sham operation group (Control group, n=10) and Model group (n=10), and the model of lower limb ASO was established. After modeling, the expression of miR-126 in arteries was detected using quantitative Real Time-Polymerase Chain Reaction (qRT-PCR), and the change in the downstream signaling pathway was examined via Western blotting. The human umbilical vein endothelial cells (HUVECs) were induced by oxidized low-density lipoprotein (Ox-LDL) to establish the model of endothelial injury, followed by detection of miR-126 expression. Then, the Luciferase assay was performed to verify the downstream target gene of miR-126. After being cultured, HUVECs were set as Control group, Ox-LDL induction group, and Ox-LDL + miR-126 inhibitor group, and the expressions of phosphorylated-protein kinase B (p-Akt) and cleaved cysteine-aspartic protease-3 (Caspase-3) were detected in the above groups.
After the establishment of the model, the expression level of miR-126 was raised in vessels, but the phosphatidylinositol 3-hydroxy kinase (PI3K)/Akt signals were weakened (p<0.01). Ox-LDL-induced endothelial cell apoptosis promoted the expression of miR-126, and the difference was statistically significant. The bioinformatics analysis results showed that PI3KR2 was a direct target of miR-126, which was also proven via the Luciferase assay. Moreover, the transfection with miR-126 inhibitor into endothelial cells suppressed Ox-LDL-induced cell apoptosis, thereby persistently activating the PI3K/Akt signaling pathway (p<0.01).
In rats with lower limb arteriosclerosis obliterans (ASO), miR-126 represses the PI3K/Akt signaling pathway to accelerate endothelial cell apoptosis.
探讨微小核糖核酸（miR）-126对下肢动脉硬化闭塞症（ASO）大鼠的影响。
将3月龄雄性Sprague-Dawley大鼠随机分为假手术组（对照组，n = 10）和模型组（n = 10），建立下肢ASO模型。建模后，采用定量实时聚合酶链反应（qRT-PCR）检测动脉中miR-126的表达，并通过蛋白质印迹法检测下游信号通路的变化。用氧化型低密度脂蛋白（Ox-LDL）诱导人脐静脉内皮细胞（HUVECs）建立内皮损伤模型，随后检测miR-126的表达。然后，进行荧光素酶报告基因检测以验证miR-126的下游靶基因。培养后，将HUVECs分为对照组、Ox-LDL诱导组和Ox-LDL + miR-126抑制剂组，检测上述各组中磷酸化蛋白激酶B（p-Akt）和裂解的半胱天冬酶-3（Caspase-3）的表达。
模型建立后，血管中miR-126的表达水平升高，但磷脂酰肌醇3-羟基激酶（PI3K）/Akt信号减弱（p<0.01）。Ox-LDL诱导的内皮细胞凋亡促进了miR-126的表达，差异具有统计学意义。生物信息学分析结果显示PI3KR2是miR-126的直接靶标，荧光素酶报告基因检测也证实了这一点。此外，向内皮细胞转染miR-126抑制剂可抑制Ox-LDL诱导的细胞凋亡，从而持续激活PI3K/Akt信号通路（p<0.01）。
在下肢动脉硬化闭塞症（ASO）大鼠中，miR-126抑制PI3K/Akt信号通路以加速内皮细胞凋亡。
