摘要
目的通过免疫组织化学法、Western blot免疫印迹法测定脑缺血再灌注大鼠脑组织内巢蛋白的表达及其相对含量水平变化,研究脑脉通、骨髓间充质干细胞移植及其联合促进神经细胞再生的作用机制。方法青年SD大鼠,清洁级,190只,雌雄各半;随机数字表法进行2次随机分组:第1次分为假手术组和手术组,手术组包括模型组、BMSCs移植组(简称移植组)、脑脉通组、BMSCs移植联合脑脉通组(简称联合组),假手术组为10只大鼠,其余各组45只;第2次对手术组按取材时间点各分为7 d、14 d、28 d组,每组各15只。参照改良的Longa法制备局灶性脑缺血动物模型,术前4天脑脉通和联合组给予脑脉通[40.5 mg/(100 g·d),40.5 mg/mL]灌胃,假手术组、模型组和移植组以同等容积的生理盐水灌胃,1次/d;术后3 h缓慢拔出线栓进行缺血后再灌注;再灌注后24 h移植组和联合组由颈内动脉给予BMSCs悬液,模型组、脑脉通组颈内动脉给予同等体积的生理盐水;每只大鼠分别于术后48 h以及术后7 d、14 d、28 d评测神经功能,术后7 d、14 d、28 d取材,应用免疫组织化学法、Western blot蛋白免疫印迹法从蛋白水平观察各组大鼠脑组织中巢蛋白的表达以及相对含量的变化,结合图像分析技术及统计学分析进行结果分析。结果正常大鼠脑组织中可见巢蛋白的少量表达;缺血再灌注损伤后大鼠脑组织中巢蛋白的表达呈现先高后低再低的特点;与模型组比较,移植、脑脉通、联合组大鼠巢蛋白的表达增加显著(P<0.05,P<0.01,P<0.01);与脑脉通组比较,移植7d组巢蛋白的表达显著增加(P<0.01);与移植组比较,脑脉通14 d组、28 d组大鼠巢蛋白阳性表达显著性增加(P<0.05);与联合组比较,其各时间点巢蛋白表达均较移植组、脑脉通组增加显著(P<0.01);同组不同时间点比较,移植7组巢蛋白表达较其14 d组、28 d组增加显著(P<0.01);脑脉通7 d组、14 d组巢蛋白表达
Objective The expression and relative content of nestin in brain tissue of rats with cerebral ischemia-reperfusion were measured by immunohistochemistry and Western blot.The mechanism of Naomaitong(脑脉通),bone marrow mesenchymal stem cell transplantation and their combination promoting nerve cell regeneration were studied.Method A total of 190 young SD rats,clean grade,half male and half female were firstly divided into sham operation group and operation group.The operation group included model group,BMSCs transplantation group(hereinafter referred to as transplantation group),Naomaitong group and BMSCs transplantation combined with Naomaitong group(hereinafter referred to as combination group).The sham operation group had 10 rats and the other groups had 45 rats.For the second time,the operation group was divided into 7 d,14 d and 28 d groups,with 15 rats in each group.The animal model of focal cerebral ischemia was prepared according to the improved Longa method.Naomaitong and the combination groups were given Naomaitong[40.5 mg/(100 g·d),40.5 mg/mL]by gavage 4 days before operation,and the sham operation group,model group and transplantation group were given the same volume of normal saline once a day.The lead bolt was slowly pulled out 3 hours after operation for ischemia-reperfusion.BMSCs suspension was given by internal carotid artery in transplantation group and combined group 24 hours after reperfusion,and the same volume of normal saline was given by internal carotid artery in model group and Naomaitong group.The neurological function of each rat was evaluated at 48 h and 7 d,14 d and 28 d after operation.The materials were taken at 7 d,14 d and 28 d after operation.The expression and relative content of nestin in brain tissue of each group were observed by immunohistochemistry and Western blot.The results were analyzed by image analysis technology and statistical analysis.Result The sham operation group had weak nestin-positive expression.The relative amount of nestin in the rats of ischemia reperf
作者
冯静静
刘昱言
刘轲
孟淑辉
刘敬霞
FENG Jingjing;LIU Yuyan;LIU Ke;MENG Shuhui;LIU Jingxia(Zhengzhou Yihe Hospital,Zhengzhou 450000,Henan,China;Third Affiliated Hospital of Henan University of Traditional Chinese Medicine,Zhengzhou 450000,Henan,China;First Affiliated Hospital of Henan University of Traditional Chinese Medicine,Zhengzhou 450000,Henan,China;Henan Provincial People's Hospital,Zhengzhou 450000,Henan,China;Ningxia Medical University,Yinchuan 750004,Ningxia,China)
出处
《辽宁中医杂志》
CAS
2021年第11期181-185,共5页
Liaoning Journal of Traditional Chinese Medicine
基金
国家自然科学基金(30772842,30973744)
关键词
缺血再灌注损伤
巢蛋白
干细胞移植
脑脉通
神经再生
ischemia-reperfusion injury
nestin
stem-cell transplantion
Naomaitong(脑脉通)
nerve regeneration
