脂肪间充质干细胞治疗脑瘫大鼠的作用机理研究  

The therapeutic mechanism of adipose mesenchymal stem cells on cerebral palsy induced by pyramidal tract injury in rats

作　　者：何正义 邹征伟 罗耀玲[1,2] 李林材 屈家阳 赖仲宏 叶俊松 HE Zhengyi;ZOU Zhengwei;LUO Yaoling;无(Subcenter for Stem Cell Clinical Translation,First Affiliated Hospital of Gannan Medical University,Ganzhou 341000,Jiangxi,PR China;Clinical Medicine Research Center,First Affiliated Hospital of Gannan Medical University,Ganzhou 341000,Jiangxi,PR China;Key Laboratory of Prevention and treatment of cardiovascular and cerebrovascular diseases,Ministry of Education,Gannan Medical University,Ganzhou 341000,Jiangxi,PR China;Key Laboratory of biomaterials and biofabrication in tissue engineering of Jiangxi Province,Gannan Medical University,Ganzhou 341000,Jiangxi,PR China;Ganzhou Key Laboratory of Stem Cell and Regenerative Medicine,Ganzhou 341000,Jiangxi,PR China;Gannan Medical University,Ganzhou 3410009 Jiangxi,PR China.)

机构地区：[1]赣南医学院第一附属医院干细胞临床转化分中心 [2]赣南医学院第一附属医院临床医学研究中心 [3]赣南医学院江西省医用组织工程材料与生物制造重点实验室 [4]赣南医学院心脑血管疾病防治教育部重点实验室 [5]赣州市干细胞与再生医学重点实验室 [6]赣南医学院,赣州341000

出　　处：《江西医药》2022年第7期681-686,689,共7页Jiangxi Medical Journal

基　　金：江西省教育厅项目,编号GJJ201535,GJJ201542,GJJ180791;江西省卫生健康委科技计划项目资助,编号202130652,202130627,20191079;心脑血管疾病防治教育部重点实验室课题,编号XN201913;赣南医学院,重点创新团队,“干细胞治疗自闭症”科技创新团队,编号TD201706;赣南医学院重点项目,编号ZD201831;赣南医学院第一附属医院,博士启动资金,编号QD066,QD076;赣南医学院第一附属医院,干细胞临床转化脑瘫专项,编号20190509-1;赣州市科技计划项目经费资助,编号202101034530。

摘　　要：目的探讨Notch信号通路在脂肪间充质干细胞治疗锥体束损伤所致脑瘫大鼠中的作用机理。方法利用脑立体定位技术在大鼠锥体束注射乙醇构建脑瘫大鼠,通过尾静脉注射大鼠脂肪间充质干细胞,通过悬吊实验、旷场实验、姿势异常、肌张力、步态活动进行脑瘫行为学评估;取大鼠锥体束注射部位的脑组织做病理组织切片进行组织病理学分析,取锥体束注射部位脑组织进行荧光定量PCR检测Notch信号通路相关基因timp1、collagen1、hes1、hey1、jag1、notch1的表达。结果与模型组比,干细胞治疗组大鼠玻璃棒悬吊时间明显增长,差异显著;姿势异常、肌张力及步行姿态得到明显改善且差异显著;组织病理学分析证实脑瘫模型组大鼠脑组织纤维束增多,而脂肪间充质干细胞治疗组纤维束明显少;脂肪间充质干细胞治疗组神经细胞数量约为脑瘫模型组的2倍,大脑神经细胞再生非常明显,脑神经细胞形态及脑组织结构破坏较脑瘫组有所改善;与模型组比,干细胞治疗组jag1、timp1、collagen1和hes1基因表达水平显著降低,hey1基因表达水平明显升高。结论脂肪间充质干细胞能够通过调控Notch信号通路促进脑瘫大鼠脑神经修复,改善脑瘫大鼠的运动功能。Objective The purpose of this study was to investigate the mechanism of adipose mesenchymal stem cells treatment on cerebral palsy induced by the injury of the vertebral tract in rats.Methods Cerebral palsy rats were established by injecting ethanol into the pyramidal tract of rats with stereotaxic technique.Rat adipose mesenchymal stem cells were injected through the tail vein.Suspension test,open field test,postural abnormalities,muscle toneand gait activity were evaluated on day6th after stem cells injection.The brain tissues at the ethanol injection site of the pyramidal tract of rats were collected for histopathological analysis.The brain tissues at the injection site of the pyramidal tract were collected to detect the expressions of Notch signaling pathway-related genes,including timp1、collagen1、hes1、hey1、jag1 and Notch1.Results Compared with model group,the suspension time of glass rod in stem cell treatment group rats were significantly increased,and the difference was significant in abnormal posture,muscle tension score and walking posture.Histopathological analysis confirmed that the number of fiber tracts in cerebral tissue of rats in the cerebral palsy model group was increased,while basically undetectable in the adipose mesenchymal stem cell treatment group.The nerve cells in the adipose mesenchymal stem cell treatment group was about two times than the cerebral palsy model group,which indicates a higher regeneration potency.Consistently,the brain nerve cell morphology and brain tissue structure of stem cell treatment group was improved compared with the cerebral palsy model group.Compared with the model group rats,the expressions of jag1,timp1,collagen1 and hes1 in the treatment group was significantly decreased,while the expression of hey1 gene was significantly increased.Conclusion Adipose mesenchymal stem cells promote cerebral nerve repair and improve motor function of cerebral palsy rats through Notch signaling pathway.

关 键 词：脑瘫 锥体束注射 脂肪间充质干细胞 神经细胞再生 NOTCH信号通路 

分 类 号：R742.3[医药卫生—神经病学与精神病学]