摘要
目的观察电针对大脑中动脉闭塞(middle cerebral artery occlusion,MCAO)后缺血再灌注(ischemia-reperfusion,I/R)不同时间点大鼠海马区内源性神经干细胞(endogenous neural stem cells,e NSCs)表达的影响,探讨电针对急性脑梗死(acute cerebral infarction,ACI)远隔损害的可能机制。方法雄性SPF级SD大鼠用线栓法制作MCAO模型,采用随机数字表法分为假手术组、脑梗死组、电针组,每组30只;假手术组仅做手术创伤,脑梗死组仅作MCAO缺血再灌注处理;电针组选取"百会"和"大椎"行电针治疗,每天1次,每次30 min;治疗后第1、7、14天分别评定神经功能缺损程度,同时在各组随机选取6只大鼠处死,分离出缺血侧海马组织,采用免疫荧光法检测海马组织e NSCs增殖与分化表达。结果 (1)大鼠神经功能缺损评分方面:与假手术组比较,脑梗死组在第1、7、14天神经功能缺损评分增高(P<0.05);与脑梗死组比较,电针组在第1、7、14天神经功能缺损评分降低(P<0.05)。(2)Brd U阳性细胞表达:与假手术比较,脑梗死组缺血侧海马CA1、CA3、DG处Brd U阳性细胞数在第1、7、14天均增多(P<0.05);与脑梗死组比较,电针组Brd U阳性细胞数均增多(P<0.05)。(3)Nestin阳性细胞表达:与假手术组比较,脑梗死组海马齿状回Nestin阳性细胞数在第1、7、14天均增多(P<0.05);与脑梗死组比较,电针组Nestin阳性表达增加,但仅第7天差异明显(P<0.05)。(4)DCX阳性细胞表达:与假手术组比较,脑梗死组第1、7天DCX阳性细胞增多(P<0.05);与脑梗死组比较,电针组第7、14天DCX阳性细胞增多(P<0.05)。(5)Neu N阳性细胞表达:与假手术组比较,脑梗死组海马齿状回Neu N阳性细胞表达在第1、7、14天均增加,但仅第14天时差异明显(P<0.05);与脑梗死组比较,电针组Neu N阳性细胞表达增多,但仅第1、14天时差异明显(P<0.05)。(6)GFAP阳性细胞表达:与假手术组比较,脑梗死组缺血侧海马齿状回GFAP阳性细胞表达在第1、7、14天均明显增多(P<0.05);与脑梗死组比较,电针组GFAP阳性细胞表达增多不显著,仅第14天时差异明显(P<0.05)。结论大鼠脑梗死后海马区存在e NSCs增殖与分化,电针治疗可促进脑梗死后神经缺损功能恢复,其机制可能与电针促进脑梗死海马区e NSCs增殖与分化,抑制e NSCs过度分化为星形胶质细胞,及早长时程促进e NSCs分化为神经元,促进神经再生等机制密切相关。
Objective To observe the effects of electroacupuncture（ EA） on hippocampal endogenous neural stem cells（ eNSCs） expression of middle cerebral artery occlusion（ MCAO） model rats after cerebral ischemiareperfusion（ I / R） at different time points,and to observe possible mechanisms of EA for keeping away fromdamage in acute cerebral infarction（ ACI）. Methods MCAO model was prepared in male SPF grade SD rats by suture method. Totally 90 rats were divided into the sham-operated group,the model group,and the EA group according to random number table,3 0 in each group. Rats in the sham-operated group only received surgical trauma. Rats in the model group only received MCAO I / R injury. Rats in the EA group received EA at Baihui（ DU2 0） and Dazhui（ DU1 4）,once per day,3 0 min each time. Nerve defects of rats were tested by neural function defect scale at day 1,7,1 4 of treatment,respectively. Meanwhile,6rats were executed randomly from each group. Their hippocampus tissues were isolated. Then the proliferation and differentiation expression of eNSCs in the hippocampus area were detected by immunofluorescence method. Results（ 1） The scores of nerve function defect scale： The scores of the model group increased at day 1,7,1 4 of treatment,being higher as compared with those of the sham-operated group（P〈0.05）.The scores of the EA group were lower than those of the model group at day 1,7,1 4 of treatment（P〈0.05）.（ 2） The expression of BrdU positive cells： Compared with the sham-operated group,the expression of BrdU positive cells in the model group were increased at day 1,7,1 4 of treatment（P〈0.05）.Compared with the model group at each time points,the expression of BrdU positive cells in the EA group were increased more at day 1,7,1 4 of treatment（P〈0.05）.（ 3） The expression of Nestin positive cells：The expression of Nestin positive cells were increased more in the model group than in the sham-operated group at day 1,7,1 4 of treatment（P〈0.05）. Compared with the model group,the expression of Nestin positive cells increased more in the EA group,but only with statistical difference at day 7 of treatment（P〈0.05）.（ 4） the expression of DCX positive cells： the expression of DCX positive cells were increased more in the model group than in the sham-operated group at day 1 and 7 of treatment（P〈0.05）. Compared with the model group,the expression of DCX positive cells were increased more in the EA group at day 7and 1 4 of treatment（P〈0.05）.（ 5） the expression of Neu N positive cells： The expression Neu N of positive cells were increased more in the model group than in the sham-operated group at day 1,7,and 1 4 of treatment,but only with statistical difference at day 1 4 of treatment（P〈0.05）. Compared with the model group,the expression of Neu N positive cells were increased more obviously, but only with statistical difference at day 1 and 1 4 of treatment（P〈0.05）.（ 6） the expression of GFAP positive cells： The expression of GFAP positive cells increased more obviously in the model group than in the sham-operated group at day 1,7,and 1 4 of treatment（P〈0.05）. Compared with the model group,the expression of GFAP positive cells were not obviously increased in the EA group,but only with statistical difference at day1 4 of treatment（P〈0.05）. Conclusions The proliferation and differentiation of eNSCs exist in the hippocampus area after cerebral I / R in MCAO model rats. EA could improve the recovery of damaged nerve function. Its possible mechanism might lie in that EA could promote the proliferation and differentiation of eNSCs in hippocampus area,inhibit excessive differentiation of eNSCs into astrocytes,promote differentiation of eNSCs into neurons,and improve regeneration of nerve cells.
出处
《中国中西医结合杂志》
CAS
CSCD
北大核心
2017年第2期198-203,共6页
Chinese Journal of Integrated Traditional and Western Medicine
基金
国家自然科学基金资助项目(No.81473470
81072947)
广东省自然科学基金重点项目(No.8152800007000001
2014A030311033)
佛山市院市合作项目(No.2014HT10004)
关键词
脑梗死
内源性神经干细胞
神经再生
电针
cerebral infarction
endogenous neural stem cell
nerve regeneration
electroacupuncture
