电针联合人脐带间充质干细胞移植对缺血性脑损伤大鼠血脑屏障及腺苷A2A受体、GSK-3β/β-catenin表达的影响  

Effects of Electro-acupuncture Combined with h UC-MSCs Transplantation on Blood-Brain Barrier,Expressions of Adenosine A2A Receptor and GSK-3β/β-catenin in Rats with Ischemic Brain Injury

作　　者：黄亮[1] 朱作磊 靖雄飞 艾威 王煜 HUANG Liang;ZHU Zuo-Lei;JING Xiong-Fei;AI Wei;WANG Yu(Xiantao First People's Hospital Affiliated to Changjiang Hosptial,Xiantao 433000 Hubei,China;Affiliated Hospital of Tianjin University of Traditional Chinese Medicine,Tianjin 300000,China)

机构地区：[1]长江大学附属仙桃市第一人民医院,湖北仙桃433000 [2]天津中医药大学附属医院,天津300000

出　　处：《广州中医药大学学报》2023年第4期935-942,共8页Journal of Guangzhou University of Traditional Chinese Medicine

基　　金：国家自然科学基金资助项目(编号:81704148)。

摘　　要：【目的】观察电针百会、风府、双侧肾俞穴联合人脐带间充质干细胞(hUC-MSCs)移植对缺血性脑损伤大鼠的脑保护作用及机制。【方法】将40只SD大鼠随机分为正常组、模型组、移植组、联合组,每组10只。除正常组,其他各组大鼠建立脑缺血再灌注损伤模型。在造模结束24 h后,移植组大鼠给予0.5 m L 2×10^(6)个hUC-MSCs细胞悬液一次性尾静脉植入,联合组在移植组治疗基础上,给予电针百会穴、风府穴、双侧肾俞穴,每次30 min,每日1次,连续针刺3周。治疗结束后,苏木素-伊红(HE)染色法观察海马组织病理形态,脱氧核糖核苷酸末端转移酶介导的缺口末端标记(TUNEL)法观察脑组织细胞凋亡情况,免疫荧光法检测脑组织腺苷A2A受体(ADORA2A)表达,免疫组织化学法检测脑组织糖原合酶激酶3β(GSK-3β)、β-连环蛋白(β-catenin)表达,Western Blot法检测脑组织跨膜蛋白闭锁蛋白(Occludin)、胞质附着蛋白(ZO-1)表达。【结果】与正常组比较,模型组脑组织细胞凋亡率升高,脑组织ADORA2A阳性表达率及GSK-3β蛋白表达水平升高,Occludin、ZO-1、β-catenin蛋白表达水平降低(P<0.05),HE染色结果显示,模型组海马组织结构明显异常;与模型组比较,移植组和联合组大鼠脑组织细胞凋亡率降低,脑组织ADORA2A阳性表达率及GSK-3β蛋白表达水平降低,Occludin、ZO-1、β-catenin蛋白表达水平升高(P<0.05),海马组织病理程度明显减轻;与移植组比较,联合组脑组织细胞凋亡率降低,脑组织ADORA2A阳性表达率及GSK-3β蛋白表达水平降低,Occludin、ZO-1、β-catenin蛋白表达水平升高(P<0.05)。【结论】电针百会、风府、双侧肾俞穴联合hUC-MSCs可改善缺血性脑损伤大鼠血脑屏障,抑制脑组织ADORA2A、GSK-3β表达,提高β-catenin表达,抑制脑组织细胞凋亡,起到脑保护作用,且作用效果优于单纯hUC-MSCs移植。Objective To observe the cerebral protective effect and mechanism of electro-acupuncture at Baihui(DU20)point,Fengfu(DU16)point and bilateral Shenshu(BL23)points combined with human umbilical cordderived mesenchymal stem cells(hUC-MSCs)transplantation on rats with ischemic brain injury. Methods Forty SD rats were randomly divided into normal group,model group,transplantation group and combination group,with 10 rats in each group. Except for the normal group,the rats in all groups were established as a model of cerebral ischemia-reperfusion injury. After 24-hour modeling,the rats in the transplantation group were given0.5 m L suspension of 2 × 10^(6)hUC-MSCs for single tail vein implantation,while the combination group was given electro-acupuncture at Baihui point,Fengfu point,and bilateral Shenshu points on the basis of treatment in the transplantation group, 30 minutes each time, once a day for consecutive 3 weeks. At the end of treatment,hippocampal histopathology was observed by hematoxylin-eosin(HE)staining,brain apoptosis was observed by deoxyribonucleotide terminal transferase-mediated nick end labeling(TUNEL)method, brain adenosine A2A receptor(ADORA2A) expression was detected by immunofluorescence, brain glycogen synthase kinase 3β(GSK-3β)and β-catenin expressions were detected by immunohistochemistry,the expressions of Occludin and zonula occludens 1(ZO-1) were detected by Western Blot. Results Compared with the normal group, the apoptosis rate of brain tissue in the model group was increased,the ADORA2A positive expression rate and GSK-3β protein expression level in brain tissue were increased,the protein expression levels of Occludin,ZO-1 andβ-catenin were decreased(P<0.05),and the HE staining results showed that the hippocampal tissue structure in the model group was significantly abnormal;compared with the model group,the apoptosis rate of brain tissue in the transplantation group and combination group was decreased,the ADORA2A positive expression rate and GSK-3β protein expression level were

关 键 词：电针 人脐带间充质干细胞(hUC-MSCs) 缺血性脑损伤 血脑屏障 腺苷A2A受体(ADORA2A) 糖原合酶激酶3β(GSK-3β) β-连环蛋白(β-catenin) 百会穴 风府穴 肾俞穴 大鼠 

分 类 号：R245[医药卫生—针灸推拿学]