Ngn2基因诱导的神经潜能骨髓间充质干细胞移植联合Janus激酶信号转导及转录激活因子通路抑制剂治疗大鼠脑缺血  

Co-transplantation of neurogin2-induced bone mesenchymal stem cells and Janus kinase-signal transducer and activators of transcription signaling pathway inhibitor(AG490)for focal cerebral ischemia/reperfusion injury in rats

作　　者：郝怀勇 吴超[1] 向金保 陈凡宇 叶翔 李永涛[1] 黄保胜 吕守华 Hao Huaiyong;Wu Chao;Xiang Jinbao;Chen Fanyu;Ye Xiang;Li Yongtao;Huang Baosheng;Lyu Shouhua(Department of Neurosurgery,the Central People′s Hospital of Tengzhou City Affiliated to Jining Medical College,Zaozhuang 277599,China;Department of General Surgery,Shanting District People′s Hospital,Zaozhuang 277200,China;Department of Neurosurgery,Sir Run Run Hospital,Nanjing Medical University,Nanjing 210006,China)

机构地区：[1]济宁医学院附属滕州市中心人民医院神经外科,枣庄277599 [2]枣庄市山亭区人民医院普通外科,277200 [3]南京医科大学附属逸夫医院神经外科,210006

出　　处：《中华实验外科杂志》2021年第9期1685-1689,共5页Chinese Journal of Experimental Surgery

基　　金：国家自然科学基金项目(81501059);济宁医学院教师科研扶持基金(JYFC2019FKJ134)。

摘　　要：目的观察Ngn2基因(Ngn2)诱导的神经潜能骨髓间充质干细胞(MSCs)和Janus激酶信号转导及转录激活因子(JAK-STAT)信号通路抑制剂AG490联合移植对大鼠局灶性脑缺血/再灌注损伤的神经保护作用。方法体外培养MSCs(南京医科大学药学院实验室培养)并将Ngn2基因转染MSCs,细胞免疫荧光观察基因转染后神经细胞标记蛋白表达情况。制备大鼠大脑中动脉栓塞(MCAO)模型。100只SD大鼠按照随机数字表达分为5组,分别设为Sham组,1%二甲基亚砜(DMSO)组、AG490组、Ngn2-MSCs+1%DMSO移植组、Ngn2-MSCs+AG490移植组。移植治疗24 h后原位缺口末端标记法(TUNEL)法检测大鼠脑缺血区细胞凋亡水平。分别于3、7、14 d对每组大鼠进行神经缺陷症状评分,14 d后处死动物,氯化三苯四唑(TTC)法染色后计算脑梗死体积,干湿重法称量脑含水量。结果 Ngn2转染MSCs后,Ngn2-MSCs细胞表现出向神经细胞分化的潜能,Ngn2-MSCs联合AG490移植治疗组中大鼠脑缺血区细胞凋亡水平(16.2±5.6,F=197.53,P<0.01)、脑梗死体积[(12.31±6.24)%,F=35.92,P<0.01]及脑含水量[(77.55±0.53)%,F=56.18,P<0.01]较其他各组明显降低,神经缺陷评分在移植后7 d较其他各组开始降低(3.31±0.46,F=14.47,P<0.01),移植治疗14 d后评分显著较其他组降低(1.63±0.35,F=22.71,P<0.01)。结论 Ngn2-MSCs联合AG490移植疗法对脑缺血再灌注损伤有明显协同神经保护作用。Objective To investigate the neuroprotective effect of combined application of neurogenin2-induced rat bone mesenchymal stem cells(Ngn2-MSCs)and Janus kinase-signal transducer and activators of transcription(JAK-STAT)signaling pathway inhibitor(AG490)on focal cerebral ischemia in rats.Methods In vitro,MSCs were isolated and cultured,and MSCs were transfected with lenti-Ngn2.In all rats,the middle cerebral artery occlusion(MCAO)model was established by occlusion of right internal carotid artery with a nylon monofilament.Totally,100 male Sprague-Dawley(SD)rats were divided into five groups according to random digits table(n=20 in each group),including Sham group,1%dimethylsurfoxide(DMSO)transplanted group,AG490 transplanted group,Ngn2-MSCs+1%DMSO transplanted group and Ngn2-MSCs+AG490 transplanted group.Cell apoptosis was examined by terminal-deoxynucleotidyl transferase mediated nick end labeling(TUNEL).The neurological defective scores were assessed at 3,7,14 day after reperfusion respectively in each experiment and all the animals were then decapitated to determine the brain infarct volume and brain water content after 14 days.Results Co-transplantation of bone MSCs and AG490 could significantly decrease the number of apoptosis cells(16.2±5.6,F=197.53,P<0.01),infarct volumes[(12.31±6.24)%,F=35.92,P<0.01],and brain water content in ischemia hemisphere[(77.55±0.53)%,F=56.18,P<0.01].The neurological defective scores in MSCs+AG490 transplanted group were significantly lower than those in other transplanted groups(7 d:3.31±0.46,F=14.47,P<0.01;14 d:1.63±0.35,F=22.71,P<0.01).Conclusion Co-transplantation of Ngn2-bone MSCs and AG490 produces a synergistic neuroprotective effect.

关 键 词：脑缺血再灌注损伤 Neurogenin2基因 Janus激酶信号转导及转录激活因子信号通路 

分 类 号：R743.3[医药卫生—神经病学与精神病学]