电针介导eNOS动员内源性EPCs促MCAO/R大鼠脑内血管再生  

Electroacupuncture promotes revascularization via eNOS mobilizing EPCs in the middle cerebral artery occlusion/reperfusion rat

作　　者：朱艳含 罗勇[1] 胥虹贝 王盼欣 

机构地区：[1]重庆医科大学附属第一医院神经内科重庆市神经病学重点实验室,重庆400016

出　　处：《中国实验动物学报》2015年第3期291-296,共6页Acta Laboratorium Animalis Scientia Sinica

基　　金：教育部"高等学校博士学科点专项科研基金"联合资助课题(20095503110001);重庆市卫生局中医药科研重点(ZY20131027);重庆市卫生局中医药科研项目(渝中医2005-B-24)

摘　　要：目的探讨内皮型一氧化氮合酶(e NOS)在电针干预下对局灶脑缺血/再灌注大鼠骨髓及外周血中内皮祖细胞(EPCs)数量变化的影响,及其促大鼠脑内血管再生的机制。方法线栓法制备大鼠局灶脑缺血/再灌注模型,针刺"百会"穴及左侧"四关"穴。100只SD雄性大鼠随机分为正常组(N组)、模型组(I/R组)、模型+电针组(I/RE组)、模型+电针+L-NAME组(I/REL组)。除N组外的各组局灶脑缺血1.5 h后分为再灌注1、2、7 d三个亚组,每个亚组10只大鼠。在各时间点采用流式细胞术检测外周血及骨髓中VEGFR2+EPCs数量,荧光定量PCR技术检测缺血皮质区VEGFR2 mRNA的表达,免疫组织化学法检测缺血皮质区VEGFR2阳性细胞表达及CD34标记的微血管计数。结果与I/R组比较,电针可明显提高骨髓及外周血中EPCs的数量(P<0.01,P<0.05),而I/REL组的VEGFR2+EPCs数量相比于I/RE组则显著降低(P<0.01)。各时间点I/RE组缺血皮质区VEGFR2阳性细胞表达,VEGFR2 mRNA表达及CD34微血管计数明显高于I/R组(P<0.01),而I/REL组与之比较则显著减少(P<0.01)。结论电针可动员局灶脑缺血/再灌注大鼠内源性EPCs促大鼠缺血脑区血管再生,该作用在e NOS被抑制后减弱,推测电针动员EPCs促血管再生的作用与e NOS的激活有关。Objective To explore the effect and mechanism of electroacupuncture on e NOS in mobilizing endothelial progenitor cells( EPCs) in rat bone marrow and peripheral blood to promote revascularization in focal cerebral ischemia / reperfusion rat. Methods A total of 100 healthy male adult Sprague-Dawley( SD) rats were randomly divided into normal group( N),model group( I / R),electroacupuncture group( I / RE) and I / RE plus L-NAME( A specific antagonist of e NOS) group( I / REL),and were further divided into 1 d,2 d and 7 d subgroups after reperfusion,10 rats in each group,in addition to the N group. The rats received filament occlusion of the right middle cerebral artery for 1. 5 hours followed by reperfusion. 'Baihui'( GV 20) /'Siguan'( Hegu LI 4 / Taichong LR 3) were selected as acupucture points. Flow cytometer was used to detect the percentage of EPCs in bone marrow and peripheral blood. The expression of VEGFR2 mRNA was tested by fluorescence quantitative PCR. Immunohistochemical staining was used to detect VEGFR2+cells and to stain the CD34+microvessels. Results Compared with the I / R group,there was a significant up-regulation of the percentage of EPCs in bone marrow and peripheral blood by electroacupuncture( P < 0. 01,P < 0. 05),but decreased by the inhibitor of eNOS( P < 0. 01). Compared with the I / R group,the VEGFR2+cells,expression of VEGFR2 mRNA and CD34+microvessels were significantly increased in the I / RE group( P < 0. 01),but decreased in the I / REL group( P < 0. 01).Conclusions Electroacupuncture can effectively mobilize EPCs to promote the revascularization in focal cerebral ischemia /reperfusion rat. This effect is attenuated by inhibitor of eNOS,suggesting that the activation of eNOS mediated by electroacupuncture may be related to mobilizing EPCs to promote the revascularization.

关 键 词：局灶脑缺血/再灌注 EPCS ENOS 电针 血管再生 脑 大鼠 

分 类 号：R245[医药卫生—针灸推拿学]