电针促进溶血卵磷脂诱导的脱髓鞘小鼠髓鞘再生修复作用机制研究  

Electroacupuncture promotes regeneration and repair of myelin sheath of corpus callosum in demyelination mice

作　　者：亢筝 邹造峰 孙婧娴 朱可蓥 姜建伟 吴根诚 汪军 KANG Zheng;ZOU Zao-feng;SUN Jing-xian;ZHU Ke-ying;JIANG Jian-wei;WU Gen-cheng;WANG Jun(Department of Integrative Medicine and Neurobiology,School of Basic Medical Sciences,Institute of Brain Science,Brain Science Collaborative Innovation Center,State Key Laboratory of Medical Neurobiology,Shanghai Medical College,Fudan University,Shanghai 200032,China)

机构地区：[1]复旦大学基础医学院中西医结合学系复旦大学脑科学研究院脑科学协同创新中心医学神经生物学国家重点实验室

出　　处：《针刺研究》2020年第1期1-7,共7页Acupuncture Research

基　　金：国家自然科学基金项目(No.81202746);上海市高峰高原项目(No.20150407);国家重点基础研究发展计划(973项目)(No.2013CB531906)

摘　　要：目的:探讨电针通过加速溶血卵磷脂(LPC)诱导的脱髓鞘小鼠小胶质细胞吞噬髓鞘碎片,促进髓鞘再生修复的作用机制。方法:C57BL/6小鼠分为正常组7只、对照组7只、模型组28只、电针组28只。采用脑数字立体定位仪在小鼠左侧胼胝体注射1μL LPC建立局部脱髓鞘小鼠模型。电针组予电针"百会""至阳"穴,造模后连续3d每天电针1次,之后每隔1d电针1次,每次30min,至造模后21d。免疫荧光染色法观察注射侧胼胝体中髓鞘碱性蛋白(MBP)、酪氨酸激酶家族Axl受体(Axl)、小胶质细胞标志物Iba1和少突胶质细胞标志物Olig2的变化;Western blot法检测注射侧胼胝体中MBP蛋白相对表达量;油红O染色观察注射侧胼胝体中脱失的髓鞘碎片变化。结果:与正常组比较,造模后5d,模型组注射侧损伤区有大量髓鞘碎片堆积,胼胝体中MBP表达下降(P<0.05),Iba1表达量呈上升趋势;造模后10d,模型组损伤区MBP表达量仍然较低(P<0.01),Iba1和Olig2阳性表达增多(P<0.001,P<0.01);造模后21d,所有指标恢复正常。与模型组比较,造模后5d,电针组MBP表达显著升高(P<0.001),髓鞘碎片大量清除,Iba1和Olig2阳性表达增多(P<0.05,P<0.001),Axl表达上升(P<0.01);造模后10d,电针组MBP表达量维持较高水平(P<0.01),Iba1表达降低(P<0.01);造模后21d,各项指标均恢复正常。结论:电针可促进小胶质细胞向髓鞘损伤部位聚集,增加损伤部位Axl表达,促进髓鞘碎片清除,加快LPC诱导的脱髓鞘动物髓鞘再生。Objective To explore the mechanism of electroacupuncture(EA)in accelerating the aggregation of microglia and promoting the remyelination at the location of demyelination.Methods C57BL/6 mice were randomly divided into4 groups:normal,control,model(LPC)and LPC+EA.The demyelination model was established by microinjection of Lysolecithin(LPC,1μL)into the left corpus callosum.EA(2 Hz/15 Hz,2-4 mA)was applied to"Baihui"(GV20)and"Zhiyang"(GV9)for 30 min,once daily for 3 days,then,once every other day for 18 days.Immuno-fluorescence staining was used to observe the expression of myelin basic protein(MBP)and Axl tyrosine kinase receptor(Axl),Iba1 and numbers of Olig2-positive oligodendrocytes in the corpus callosum.Western blot was employed to detect the expression of MBP in the corpus callosum,and Oil Red O staining was used to observe changes of number of myelin pieces.Results Following modeling,the expression levels of MBP on day 5 and 10 after modeling were significantly decreased(P<0.05,P<0.01),Iba1 expression and Olig2-positive oligodendrocyte numbers on day 10 apparently increased(P<0.001,P<0.01).On day 21 after modeling,the levels of the above mentioned indexes returned to normal.After EA intervention,the levels of MBP expression on day 5 and 10,Axl,Iba1 protein expression and Olig2-positive oligodendrocyte numbers on day 5 were markedly increased(P<0.001,P<0.01,P<0.05),while Iba1 expression on day 10 was considerably decreased in comparison with the model group(P<0.01).Oil Red O staining showed that on day 5 after modeling,the number of red lipid droplets were obviously increased in the corpus callosum tissue on the injection side,and apparently reduced in the EA group,suggesting a clearance of the accumulated myelin fragments by EA.Conclusion EA intervention may reduce myelin debris and promote the aggregation of microglial cells and oligodendrocytes to the injured site,accelerate the myelin regeneration and up-regulate the expression of MBP and Axl of corpus callosum in demyelination mice.

关 键 词：髓鞘再生 电针 胼胝体 少突胶质细胞 小胶质细胞 

分 类 号：R245.97[医药卫生—针灸推拿学]