miR-23a-3p通过PTEN/PI3K/AKT通路参与ox-LDL诱导的人脐静脉内皮细胞自噬和凋亡的影响OA
Effects of miR-23a-3p on autophagy and apoptosis of human umbilical vein endothelial cells induced by ox-LDL via PTEN/PI3K/AKT Pathway
目的 探讨 miR-23a-3p 通过磷酸酶-张力蛋白同源物(PTEN)/磷脂酰肌醇-3-激酶(PI3K)/蛋白激酶 B(AKT)通路对氧化型低密度脂蛋白(ox-LDL)诱导的人脐静脉内皮细胞(HUVEC)自噬与凋亡的影响及其可能的机制,为miR-23a-3p在动脉粥样硬化(AS)中的作用研究提供理论支持.方法 采用 50 mg/L ox-LDL诱导HUVEC建立体外细胞模型(ox-LDL HUVEC,简称ox-LDL),荧光定量PCR(qRT-PCR)检测miR-23a-3p的表达水平,CCK8 检测细胞活力,流式细胞术检测细胞的凋亡情况,生物信息学及双荧光素酶报告基因实验验证 miR-23a-3p靶向基因PTEN,免疫蛋白印迹法(WB)检测靶基因PTEN、自噬相关蛋白LC3Ⅱ/LC3Ⅰ、自噬底物蛋白P62、PI3K/AKT通路相关蛋白的表达水平.结果 双荧光素酶报告基因实验结果显示miR-23a-3p可靶向调控PTEN.经ox-LDL诱导的HUVEC中miR-23a-3p表达降低(P<0.05).miR-23a-3p过表达抑制了ox-LDL诱导的HUVEC自噬,表现为PTEN、LC3Ⅱ/LC3Ⅰ蛋白表达量减少,P62 蛋白表达量增加(P<0.05),同时提升细胞活力、抑制细胞凋亡率(P<0.05);而miR-23a-3p抑制物组结果则相反(P<0.05).敲低 PTEN可促进 ox-LDL诱导的 HUVEC中 P-PI3K/PI3K、P-AKT/AKT蛋白表达增加(P<0.05),抑制自噬(P<0.05),过表达PTEN结果则相反(P<0.05).结论 miR-23a-3p通过靶向调控PTEN抑制ox-LDL诱导的HUVEC自噬,促进HUVEC活力并抑制细胞凋亡,其机制可能与激活PI3K/AKT通路蛋白表达有关,miR-23a-3p可能成为AS的潜在治疗新靶点.
Objective To investigate the effects of miR-23a-3p on autophagy and apoptosis in human umbilical vein en-dothelial cell(HUVEC)induced by oxidized low-density lipoprotein(ox-LDL)via phosphatase and tensin homolog(PTEN)/phosphatidylinositol-3-kinase(PI3K)/protein kinase B(AKT)pathway,and to explore its possible mechanisms,so as to provide theoretical support for the role of miR-23a-3p in atherosclerosis(AS).Methods An in vitro cell model of HUVEC was established by induction with 50 mg/L ox-LDL(ox-LDL HUVEC,abbreviated as ox-LDL).The expression lev-el of miR-23a-3p was detected by quantitative real-time PCR(qRT-PCR).Cell viability was assessed using the CCK8 as-say,and cell apoptosis was measured by flow cytometry.Bioinformatics and dual-luciferase reporter assays were used to veri-fy that miR-23a-3p targeting the gene PTEN.Western blot(WB)was employed to detect the expression levels of the target gene PTEN,autophagy-related proteins LC3Ⅱ/LC3Ⅰ,autophagy substrate protein P62,and proteins related to the PI3K/AKT pathway.Results The dual-luciferase reporter assay results showed that miR-23a-3p could target and regulate PTEN.In HUVEC induced by ox-LDL,the expression of miR-23a-3p decreased(P<0.05).The overexpression of miR-23a-3p in-hibited ox-LDL-induced autophagy in HUVEC,manifested as reduced expression of PTEN and LC3Ⅱ/LC3Ⅰproteins,and increased expression of P62 protein(P<0.05).Simultaneously,it enhanced cell viability and inhibited the apoptosis rate(P<0.05);in contrast,the results in the miR-23a-3p inhibitor group were opposite(P<0.05).The knockdown of PTEN promoted the increase in P-PI3K/PI3K and P-AKT/AKT protein expression in ox-LDL-induced HUVEC(P<0.05),inhibi-ted autophagy(P<0.05),while the overexpression of PTEN had the opposite effects(P<0.05).Conclusion miR-23a-3p inhibits autophagy in ox-LDL-induced HUVEC by targeting PTEN,enhances cell viability,and suppresses cell apoptosis.Its mechanism may be related to the activation of PI3K/AKT pathway protein expression,and miR-23a-3p may serve as a poten-tial therapeutic target for AS.
林金蕊;何则沂;丘金浪;陈娜君
福建省福州市中医院检验科,福建 福州 350001福建省福州市中医院检验科,福建 福州 350001福建省福州市中医院检验科,福建 福州 350001福建省福州市中医院检验科,福建 福州 350001
医药卫生
miR-23a-3p氧化型低密度脂蛋白人脐静脉内皮细胞磷酸酶-张力蛋白同源物自噬凋亡磷脂酰肌醇-3-激酶/蛋白激酶B信号通路动脉粥样硬化
miR-23a-3poxidized low-density lipoprotein(ox-LDL)human umbilical vein endothelial cell(HUVEC)phosphatase and tensin homolog(PTEN)autophagyapoptosisphosphatidylinositol-3-kinase(PI3K)/protein kinase B(AKT)signaling pathwayatherosclerosis(AS)
《右江医学》 2026 (2)
122-130,9
福州市卫生健康系统科技计划项目(2022-S-wq19)
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