miR-152-3p通过AKT/GSK-3β/Nrf2信号通路促进脑出血大鼠神经元铁死亡  

miR-152-3p via AKT/GSK-3β/Nrf2 signaling pathway promoting neuronal ferroptosis in rats with cerebral hemorrhage

作　　者：汪赟辉 张笑锋 WANG Yunhui;ZHANG Xiaofeng(Department of Neurosurgery,Deqing People’s Hospital(Zhejiang University Affiliated Shaw Hospital Deqing Campus),Huzhou 313200,China)

机构地区：[1]德清县人民医院(浙江大学附属邵逸夫医院德清院区)神经外科,浙江湖州313200

出　　处：《温州医科大学学报》2024年第4期287-295,301,共10页Journal of Wenzhou Medical University

基　　金：湖州市自然科学基金项目(2022YZ55)。

摘　　要：目的:探讨miR-152-3p通过AKT/GSK-3β/Nrf2信号通路促进脑出血(ICH)大鼠神经元铁死亡的机制。方法:使用大鼠神经元细胞为研究对象,采用氧合血红蛋白(oxyHb)刺激神经元细胞构建ICH体外细胞模型,分别将miR-152-3p inhibitor和(或)PIK3CA抑制剂分别处理细胞,采用流式细胞术观察细胞凋亡情况,RT-qPCR、Western blot检测相关基因和蛋白表达情况;并用细胞免疫荧光观察Nrf2蛋白的入核率;用双荧光素酶靶标实验验证miR-152-3p与PIK3CA的关系。结果:将miR-152-3p inhibitor转染的细胞构建ICH细胞模型后,细胞凋亡率明显降低(P<0.01);SLC7A11、GPx4、PIK3CA、Nrf2蛋白表达水平以及p-AKT/AKT比率显著升高而GSK-3β蛋白表达水平显著降低(P<0.01);同时,Nrf2蛋白入核量也显著升高(P<0.01);而此作用在使用PIK3CA抑制剂干预后,miR-152-3p inhibitor的改善效果消失;荧光素酶靶标实验证实,miR-152-3p可靶向调控PIK3CA。结论:miR-152-3p通过AKT/GSK-3β/Nrf2信号通路促进ICH大鼠神经元铁死亡。Objective:To explore how miR-152-3p influences neuronal ferroptosis in rats with intracerebral hemorrhage(ICH)through the Akt/GSK-3β/Nrf2 signaling pathway.Methods:Rat neuronal cells were used to create an in vitro ICH model,induced by oxygenated hemoglobin(oxyHb)stimulation.These cells were treated with either a miR-152-3p inhibitor,a PIK3CA inhibitor,or both.Cell apoptosis was assessed using flow cytometry.The expression level of relevant genes and proteins was measured through RT-qPCR and Western blot.Nrf2 protein nuclear translocation was examined via cellular immunofluorescence.Additionally,a dual-luciferase reporter assay was conducted to investigate the interaction between miR-152-3p and PIK3CA.Results:In the ICH cell model,transfection with the miR-152-3p inhibitor significantly reduced apoptosis rate(P<0.01).This treatment also notably increased the expression levels of SLC7A11,GPx4,PIK3CA,and Nrf2 proteins,as well as the p-AKT/AKT ratio,while decreasing GSK-3βprotein levels(P<0.01).Furthermore,Nrf2 protein nuclear translocation was significantly enhanced(P<0.01).However,these effects were mitigated upon PIK3CA inhibitor treatment.The luciferase reporter assay confirmed that miR-152-3p could target and regulate PIK3CA.Conclusion:miR-152-3p,through the Akt/GSK-3β/Nrf2 signaling pathway,promotes neuronal ferroptosis in rats experiencing intracerebral hemorrhage.

关 键 词：miR-152-3p AKT/GSK-3β/Nrf2信号通路 铁死亡 

分 类 号：R96[医药卫生—药理学]