NT-3基因修饰骨髓间充质干细胞移植对七氟醚诱导老龄小鼠认知功能障碍的保护作用  

Protective effect of NT-3 gene modified bone marrow mesenchymal stem cell transplantation on sevoflurane induced cognitive dysfunction in aged mice

作　　者：杜薇 张晓敏 谢昕 李松泽 黄泽清 DU Wei;ZHANG Xiao-min;XIE Xin;LI Song-ze;HUANG Ze-qing(Department of Anesthesiology,Cancer Hospital of China Medical University,Liaoning Cancer Hospital&Insitute,Shenyang 110042,China)

机构地区：[1]中国医科大学肿瘤医院辽宁省肿瘤医院麻醉科,辽宁沈阳110042

出　　处：《解剖科学进展》2024年第2期149-152,共4页Progress of Anatomical Sciences

基　　金：辽宁省自然科学基金(2023-BS-044)。

摘　　要：目的探究NT-3基因修饰骨髓间充质干细胞(BMSCs)移植对七氟醚诱导老龄小鼠认知功能障碍的保护作用及机制。方法分离培养小鼠BMSCs,病毒转染建立NT-3基因修饰的BMSCs,Western blot检测BMSCs NT-3蛋白表达,CCK-8检测BMSCs增殖能力,免疫荧光染色方法检测BMSCs神经元标志物NeuN表达。将40只12月龄C57BL/6老年小鼠随机分为对照组、七氟醚组、骨髓间充质干细胞组和NT-3基因修饰骨髓间充质干细胞组,每组10只。TUNEL实验检测小鼠海马组织细胞凋亡;免疫荧光检测小鼠海马组织神经元标志物TH;Morris水迷宫实验检测小鼠学习和记忆能力;Western blot检测小鼠海马组织β-catenin和p-GSK-3β蛋白表达水平。结果成功建立NT-3基因修饰的BMSCs,NT-3基因修饰增加BMSCs增殖能力,上调BMSCs NeuN表达。NT-3基因修饰骨髓间充质干细胞移植抑制七氟醚麻醉的小鼠海马组织细胞凋亡,促进小鼠海马组织神经元存活,增加小鼠学习和记忆能力,上调小鼠脑组织β-catenin和p-GSK-3β蛋白表达水平。结论NT-3基因修饰的骨髓间充质干细胞移植能通过激活Wnt/β-catenin信号通路改善七氟烷麻醉的老龄小鼠认知功能障碍。Objective To explore the protective effect of NT-3 gene modified bone marrow mesenchymal stem cells(BMSCs)transplantation on sevoflurane induced cognitive dysfunction in aged mice and its mechanism.Methods Mouse BMSCs were isolated and cultured,and NT-3 gene modified BMSCs were established by lentivirus transfection.The expression of NT-3 protein was detected by Western blot.The proliferation ability of BMSCs was detected by CCK-8.The expression of neuron marker NeuN of BMSCs was detected by immunofluorescence.Forty 12-month-old C57BL/6 aged mice were randomly divided into control group,sevoflurane group,bone marrow mesenchymal stem cell group and NT-3 gene modified bone marrow mesenchymal stem cell group,with 10 mice in each group.TUNEL assay was used to detect the apoptosis of hippocampal cells of mice.The neuronal marker TH in mouse hippocampus was detected by immunofluorescence.The learning and memory ability of mice was detected by morris water maze test.The expressions ofβ-catenin and p-GSK-3βin the hippocampus of mice were detected by Western blot.Results BMSCs modified with NT-3 gene were successfully established.NT-3 gene modification increased the proliferation ability of BMSCs and up-regulated the NeuN expression of BMSCs.NT-3 gene modified bone marrow mesenchymal stem cell transplantation inhibited hippocampal cell apoptosis,promoted hippocampal neuron survival,increased learning and memory ability,and up-regulatedβ-catenin and p-GSK-3βprotein expression levels in brain of sevoflurane anesthesia mice.Conclusion NT-3 gene modified bone marrow mesenchymal stem cell transplantation can improve cognitive dysfunction in sevoflurane anesthesia aged mice by activating the Wnt/β-catenin signaling pathway.

关 键 词：神经营养因子3 骨髓间充质干细胞 七氟醚 认知功能障碍 WNT/Β-CATENIN信号通路 

分 类 号：R749.6[医药卫生—神经病学与精神病学]