李思^1,,
孙经淞^2,,
1. 大连医科大学附属第一医院 检验科，辽宁 大连 116011
2. 大连医科大学附属第一医院 创伤骨科，辽宁 大连 116011

基金项目: 辽宁省博士科研启动基金（2020-BS-193）


详细信息 作者简介: 李 思（1994-），女，主管技师。E-mail：562275050@qq.com


通讯作者: 孙经淞，副主任医师。E-mail：791566418@qq.com 中图分类号: R994


摘要:目的探讨骨髓间充质干细胞（BMSCs）移植对正己烷中毒大鼠脊髓神经组织的修复作用。方法将健康成年雄性SD大鼠随机分成5组，每组20只。分别为：对照组（control）、干细胞对照组（BMSCs）、染毒组（HD）、自身修复组（HD+NS）和干细胞干预染毒组（HD+BMSCs）。对照组腹腔注射生理盐水，10周后处死动物。BMSCs组腹腔注射生理盐水5周后，行尾静脉干细胞移植术，5周后处死动物。HD组按400 mg/kg剂量，对大鼠腹腔注射2，5-己二酮（2，5-hexanedione，HD），每周1次，持续注射5周后，处死动物。HD+NS组对大鼠腹腔注射HD 5周后，经尾静脉注射生理盐水1 mL，5周后处死动物。HD+BMSCs组对大鼠腹腔注射HD 5周后，经尾静脉行干细胞移植，5周后处死动物。实验5周后观察大鼠的一般状态并进行步态评分。透射电镜观察处死后大鼠脊髓神经组织超微结构。MAP-2和SMI312双标记免疫荧光染色和LFB染色观察大鼠脊髓神经组织轴突和髓鞘变化。结果HD+NS组大鼠一般状态差、精神萎靡、无法站立、呈瘫痪状，步态评分为（4±0.38）分，属严重异常。HD+BMSCs组大鼠在BMSCs移植2周后，大鼠精神状态明显提升、摄食量升高，随着移植时间的延长，大鼠步态逐渐改善。电镜结果显示，HD中毒大鼠移植BMSCs后，脊髓神经组织可见新生轴突。SMI312和MAP2免疫荧光双染可见，BMSCs移植后神经元的SMI312免疫荧光强度明显增强。髓鞘LFB染色结果显示HD中毒大鼠脊髓组织髓鞘脱失、板层疏松，而BMSCs移植后，大鼠脊髓神经组织髓鞘结构趋于正常。结论BMSCs移植可显著改善HD中毒大鼠的神经行为、修复HD中毒大鼠脊髓神经组织受损的轴突和髓鞘。
关键词: 正己烷/
间充质干细胞/
脊髓神经组织/
移植


Abstract:ObjectiveTo investigate the repair effect of transplantation of mesenchymal stem cells (BMSCs) on spinal cord nerve tissue in hexane-poisoned rats.MethodsHealthy adult male SD rats were randomly divided into five groups of 20 rats each: control group, BMSCs group, HD group, HD+NS group and HD+BMSCs group. In the control group, saline was injected intraperitoneally; in BMSCs group, saline was injected intraperitoneally for 5 weeks, followed by tail vein stem cell transplantation for 5 weeks; in HD group, 2, 5-hexanedione (HD) 400 mg/kg was injected once a week, intraperitoneally for 5 weeks; in HD+NS group, HD was injected intraperitoneally for 5 weeks, followed by 1 mL of saline via tail vein for 5 weeks; in HD+BMSCs group, rats were injected with HD intraperitoneally for 5 weeks and then transplanted with stem cells through the tail vein for 5 weeks. The general status of the rats was observed and gait scores were performed after 5 weeks. The ultrastructure of rat spinal cord nerve tissue was studied by transmission electron microscopy, and immunofluorescence staining and LFB staining were used to observe axonal and myelin changes in rat spinal cord nerve tissue.ResultsThe rats in HD+NS group showed bad general status, depression, inability to stand, and paralysis, and the gait score was 4±0.38, indicating a serious abnormality. After 2 weeks of transplantation, the status of the rats was significantly improved, the food intake was increased, and the gait gradually improved in association with time. Electron microscopic results showed that new axons were visible in the spinal cord nerve tissue of HD-intoxicated rats after transplantation of BMSCs. Double staining of SMI312 and MAP2 immunofluorescence displayed that the SMI312 immunofluorescence intensity of neurons was significantly enhanced. LFB staining showed myelin loss and laminar sparing in the rat spinal cord tissue, while myelin structure of the rat spinal cord nerve tissue tended to be normal after transplantation of BMSCs.ConclusionBMSCs transplantation can significantly improve the neuro behavior and repair the damaged axons and myelin sheaths in the spinal cord of HD-intoxicated rats.
Keywords:n-hexane/
mesenchymal stem cells/
spinal cord neural tissue/
transplantation

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