电针通过eNOS促进局灶脑缺血/再灌注大鼠脑缺血皮质区血管再生  

Effects of electroacupuncture on angiogenesis via eNOS in rat ischemic cerebral cortex after focal cerebral ischemia/reperfusion

作　　者：胥虹贝 罗勇[1] 朱艳含 李娟[1] 王盼欣 张莹[1] 

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：《中国组织化学与细胞化学杂志》2015年第5期381-387,共7页Chinese Journal of Histochemistry and Cytochemistry

基　　金：国家临床重点专科建设项目经费资助(卫办医政函[2012]649号);重庆市卫生局中医药科研重点项目(ZY20131027);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)

摘　　要：目的观察电针对局灶脑缺血/再灌注大鼠大脑缺血皮质区e NOS、MMP-9表达及CD34+微血管密度变化的影响,探讨电针促进大脑缺血皮质区血管再生的机制。方法 120只SD雄性大鼠随机分为正常组(NC组)、模型组(I/R组)、模型+电针组(I/RE组)、模型+电针+e NOS特异性拮抗剂L-NAME组(I/REL组)。采用线栓法制备大鼠局灶脑缺血/再灌注模型,缺血90min后将I/R、I/RE和I/REL组分各为再灌注1d、3d和7d三个亚组。取大鼠"百会"穴(GV 20)及左侧"四关"穴(合谷LI 4/太冲LR 3)为电针穴位,刺激时间为20min/d,最长持续7d。采用免疫组化法检测各组大脑缺血皮质区e NOS、MMP-9和CD34表达,荧光定量PCR技术检测大脑缺血皮质区e NOS mRNA表达。结果与NC组比较,I/R、I/RE与I/REL组大脑缺血皮质区e NOS mRNA及蛋白表达随再灌注时间延长呈进行性增加,与I/R组比较,I/RE组各时间点e NOS mRNA及蛋白表达均显著升高,在L-NAME作用下,I/REL组e NOS mRNA及蛋白表达明显低于I/RE组。再灌注后1d、3d,I/RE组MMP-9表达明显低于I/R和I/REL组,7d时I/RE组MMP-9表达较I/R、I/REL组升高。I/RE组各时间点CD34+微血管密度升高较I/R组显著,I/REL组微血管密度明显低于I/RE组。结论电针可通过上调局灶脑缺血/再灌注大鼠脑缺血皮质区e NOS和MMP-9表达,促进血管再生。Objective To explore the mechanism of electroacupuncture in promoting angiogenesis in ischemic cortex by investigating the effect of electroacupuncture on eN OS,MMP-9 and CD34+microvessels in rats after focal cerebral ischemia / reperfusion.Methods A total of 120 healthy male adult Sprague-Dawley( SD) rats were randomly assigned to normal control group( NC),model group( I / R),electroacupuncture group( I / RE) and I / RE plus L-NAME( A specific antagonist of eN OS) group( I / REL). The model of focal cerebral ischemia / reperfusion was prepared by the thread embolism method,and all groups were divided into 1d,3d and 7d subgroups after reperfusion except for the NC group. Electroacupuncture was given at 'Baihui'( GV20) and 'Siguan'on the left side( Hegu LI 4 /Taichong LR 3) acupoints for 20 min per day. Immunohistochemical method was used to detected the expression of eN OS,MMP-9 and CD34+microvessels. The mRNA expression of eN OS was measured by fluorogenic quantitative PCR. Results Compared with those of the NC group,the protein and mRNA expressions of eN OS in the other 3 groups were significantly upregulated with lengthening of reperfusion time,and obviously higher in the I / RE group than in I / R and I / REL groups. The protein expression of MMP-9in the I / RE group was lower than that in I / RE and I / REL groups at 1d and 3d,but higher in the I / RE group than in the other 3groups at 7d after reperfusion. The amount of CD34+microvessels in the I / RE group was higher than that of I / R and I / REL groups.Conclusion Electroacupuncture can effectively promote angiogenesis by upregulating the expression of eN OS,MMP-9 in the ischemic cerebral cortex of rats after focal cerebral ischemia / reperfusion.

关 键 词：局灶脑缺血/再灌注 电针 L-NAME ENOS MMP-9 血管再生 

分 类 号：R245.97[医药卫生—针灸推拿学]