miR-132-3p靶向SIRT1对施旺细胞损伤及再生的影响  

Effect of miR-132-3p targeting SIRT1 on repair and regeneration after sciatic nerve injury

作　　者：车敏[1] 陈星宇 王岩峰[3] 李良满[3] 张辉[1] CHE Min;CHEN Xing-yu;WANG Yan-feng;LI Liang-man;ZHANG Hui(Department of Hand Surgery,Afiliated Central Hospital of Shenyang Medical College,Shenyang 110024;Trauma Repair Surgery Department,Bazhou People's Hospital,Korla City,Bayingolin Mongolian Autonomous Prefecture,Korla 841009;Department of Orthopedic Surgery,The First Affiliated Hospital of China Medical University,Shenyang 110001,China)

机构地区：[1]沈阳医学院附属中心医院手外一科,辽宁沈阳110024 [2]新疆巴音郭楞蒙古自治州库尔勒市巴州人民医院创伤修复外科,新疆库尔勒841009 [3]中国医科大学附属第一医院骨科,辽宁沈阳110001

出　　处：《解剖科学进展》2024年第2期161-164,169,共5页Progress of Anatomical Sciences

基　　金：国家自然基金(8197080581);辽宁省自然科学基金(2020-MS-146);辽宁省教育厅高校基本科研面上项目(LJKMZ20221781);辽宁省教育厅科学研究面上项目(LJKZ0745)。

摘　　要：目的探究miR-132-3p靶向SIRT1对施旺细胞增殖、凋亡及神经再生相关蛋白表达的影响。方法通过生物信息学分析miR-132-3p与SIRT1 mRNA的潜在结合位点,双荧光素酶报告基因实验验证二者结合;原代培养施旺细胞,RT-qPCR和Western blot检测转染miR-132-3p模拟物或抑制剂对施旺细胞SIRT1表达的影响;将SIRT1过表达质粒转染过表达miR-132-3p的施旺细胞,Western blot检测细胞SIRT1、GAP-43和MBP表达,CCK-8检测细胞增殖活性,流式细胞术检测细胞凋亡率。结果miR-132-3p靶向结合SIRT1;转染miR-132-3p模拟物降低施旺细胞SIRT1表达,转染miR-132-3p抑制剂增加施旺细胞SIRT1表达;转染miR-132-3p模拟物降低施旺细胞增殖活性并增加细胞凋亡率,降低细胞GAP-43和MBP表达,过表达SIRT1增加转染miR-132-3p模拟物的施旺细胞增殖活性并降低细胞凋亡率,增加细胞GAP-43和MBP表达。结论miR-132-3p通过靶向抑制SIRT1表达抑制施旺细胞增殖并促进细胞凋亡。Objective To explore the effects of miR-132-3p targeting SIRT1 on Schwann cell proliferation,apoptosis,and expression of nerve regeneration related proteins.Methods The potential binding sites of miR-132-3p and SIRT1 mRNA were analyzed by bioinformatics,and the double luciferase reporter gene experiment was used to verify their binding.Primary culture of Schwann cells,RT-qPCR and Western blot were used to detect the effect of miR-132-3p mimetics or inhibitors transfected on the expression of SIRT1 in Schwann cells.The SIRT1 over-expression plasmid was transfected into Schwann cells with miR-132-3p over-expression,and the expressions of SIRT1,GAP-43 and MBP were detected by Western blot.Cell proliferation activity was detected by CCK-8,and the apoptosis rate was detected by flow cytometry.Results miR-132-3p targeted SIRT1,miR-132-3p mimics decreased SIRT1 expression in Schwann cells,and miR-132-3p inhibitors increased SIRT1 expression in Schwann cells.Transfection of miR-132-3p mimics decreased Schwann cell proliferation activity and increased cell apoptosis rate,and decreased the expressions of GAP-43 and MBP.Overexpression of SIRT1 increased Schwann cell proliferation activity and decreased cell apoptosis rate,and increased the expression of GAP-43 and MBP in Schwann cells transfected with miR-132-3p mimics.Conclusion miR-132-3p inhibits Schwann cell proliferation and promotes apoptosis by inhibiting SIRT1 expression.

关 键 词：miR-132-3p 坐骨神经损伤 施旺细胞 神经再生 神经修复 SIRT1 

分 类 号：R651.3[医药卫生—外科学]