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
车敏 1陈星宇 2王岩峰 3李良满 3张辉1
作者信息
- 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
- 折叠
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S1RT1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.
Abstract
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 S1RT1,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/SIRT1Key words
miR-132-3p/sciatic nerve injury/Schwann cells/nerve regeneration/nerve repair/SIRT1引用本文复制引用
基金项目
国家自然科学基金(8197080581)
辽宁省自然科学基金(2020-MS-146)
辽宁省教育厅高等学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)
出版年
2024
NETL
NSTL
万方数据