骨髓间充质干细胞对树脂毒素介导的小鼠神经病理性疼痛模型镇痛作用及其机制  

The analgesic effect of bone mesenchymal stem cells transplantation in resiniferatoxin-induced neuropathic pain in mice

作　　者：李楠琦 徐雯雯 汤洋 安珂[1] 万丽[2] LI Nan-Qi;XU Wen-Wen;TANG Yang;AN Ke;WAN Li(Department of Anesthesiology,The First Affiliated Hospital,Sun Yat-sen University,Guangzhou 510080,China;Department of Pain Management,The Second Affiliated Hospital,Guangzhou Medical University,State Key Clinical Specialty in Pain Medicine,Guangzhou 510260,China)

机构地区：[1]中山大学附属第一医院麻醉科,广州510080 [2]广州医科大学附属第二医院疼痛科,广州510260

出　　处：《中国疼痛医学杂志》2020年第9期660-667,共8页Chinese Journal of Pain Medicine

基　　金：国家自然科学基金(81171468;81771182);广东省自然科学基金(2018A03031300297)。

摘　　要：目的:探究骨髓间充质干细胞(bone mesenchymal stem cells, BMSC)对树脂毒素(resiniferatoxin,RTX)介导的神经病理性疼痛小鼠的镇痛作用及其机制。方法:36只C57BL/6小鼠随机均分为3组(每组12只):溶剂对照组(Ctrl组):小鼠腹腔注射溶剂、RTX神经痛组(RTX组):腹腔注射树脂毒素建立RTX神经痛模型、骨髓间充质干细胞移植组(BMSC组):骨髓间充质干细胞移植组于造模后5天经尾静脉输注骨髓间充质干细胞。在RTX注射后1、5、10、15、20、25、30天分别测量各组小鼠机械痛阈和热痛阈。在模型建立后30天分别对3组小鼠进行取材,应用蛋白印迹检测p38 MAPK表达量的变化,应用免疫荧光技术检测脊髓背角小胶质细胞活化及BMSC归巢情况,同时应用高尔基染色观察3组小鼠脊髓背角树突棘可塑性变化。结果:RTX组小鼠机械痛阈值在第5天开始下降,第10天及以后机械痛阈值与对照组相比,均有显著性差异(P <0.05);BMSC组机械痛阈值增加,BMSC注射后第20天差异有显著性意义(P <0.05)。RTX处理后脊髓小胶质细胞标记物CD11b及p38 MAPK的表达显著高于对照组,RTX组中小胶质细胞与p38 MAPK双染高度共定位。BMSC注射后小胶质细胞激活减弱,p38 MAPK蛋白表达降低。结论:骨髓间充质干细胞移植在RTX介导的神经病理性疼痛模型中可定向迁移至脊髓背角部位,可通过减轻小胶质细胞活化和p38 MAPK通路激活发挥镇痛作用。Objective:To elucidate the analgesic effect of bone mesenchymal stem cells(BMSC)transplantation on resiniferatoxin(RTX)induced neuropathic pain in mice and their potential mechanism related to microglia activation.Methods:Thirty-six C57BL/6 mice were randomly divided into 3 groups(12 in each group).Control group:intraperitoneal injection of vehicle;RTX neuralgia group:establishment of RTX neuralgia model by intraperitoneal injection of resinferatoxin;Bone marrow mesenchymal stem cell transplantation group(BMSC):1×10^6 BMSC was infused through tail vein 5 days after modeling.Mechanical threshold and thermal latency of mice in each group were measured at day 1,5,10,15,20,25 and 30 after administration.Three groups of mice were sampled 30 days after the establishment of the model.Changes of p38 MAPK expression were detected by western blot,microglia activation in spinal dorsal horn and BMSC tracing were detected by immunofluorescence,and plasticity of dendritic spines in spinal dorsal horn was observed by Golgi staining.Results:The mechanical withdrawal threshold(MWT)of mice in RTX group began to decline from day 5,since day 10,the difference between RTX and control group was significant(P<0.05).Mechanical withdrawal threshold increased in BMSC group,and the difference was significant at day 20 after BMSC injection(P<0.05).After RTX treatment,the expression of microglia markers CD11b and p38 MAPK in the spinal cord upregulated than those in the control group,and CD11b in RTX group were co-localized with p38 MAPK.After BMSC injection,the activation of microglia was attenuated and p38 MAPK expression was decreased.Conclusion:Systemic transplantation of BMSC can be directed to the spinal cord dorsal horn in RTX induced neuropathic pain model,and it can produce analgesic effect by reducing microglial activation and p38 MAPK level in RTX treated mice.

关 键 词：骨髓间充质干细胞移植 树脂毒素 神经病理性疼痛 小胶质细胞激活 

分 类 号：R741[医药卫生—神经病学与精神病学] R-332[医药卫生—临床医学]