电针对局灶脑缺血/再灌注大鼠骨髓及外周血CD34^+内皮祖细胞的影响  

Effect of Electroacupuncture on CD 34^+ Endothelial Progenitor Cell Counts in Bone Marrow and Peripheral Blood in Focal Cerebral Ischemia/Reperfusion Rats

作　　者：谢宸宸[1] 罗勇[1] 庞月珊[1] 高祥[2] 李满[1] 汶海琪 陈瑞芳[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 [2]成都铁路分局医院肾内科,成都610081

出　　处：《针刺研究》2014年第6期437-442,共6页Acupuncture Research

基　　金：重庆市卫生局中医药科研重点项目(No.ZY 20131027);重庆市卫生局中医药科研项目(渝中医2005-B-24);教育部"高等学校博士学科点专项科研基金"联合资助课题(No.20095503110001)

摘　　要：目的:观察电针对局灶脑缺血/再灌注大鼠骨髓及外周血CD 34+内皮祖细胞(EPCs)数量、磷酸化蛋白激酶B(p-AKT)表达的影响,探讨电针动员骨髓CD 34+EPCs入血,促进脑缺血修复的机制。方法:雄性SD大鼠随机分为假手术组、模型组、电针组。采用线栓法制备局灶脑缺血/再灌注大鼠模型,局灶脑缺血2h后将各组分为再灌注12、24、48h3个时间点,每个时间点12只大鼠。取大鼠"百会"穴及左侧"四关"穴("合谷"/"太冲")为电针穴位,刺激时间30min,每日1次。采用神经行为学评分法评价大鼠神经功能缺损情况,流式细胞术检测骨髓及外周血CD 34+EPCs数量,Western blot法检测骨髓p-AKT蛋白的表达。结果:模型组再灌注后各时间点均有不同程度的神经功能缺损,电针可明显改善大鼠再灌注后48h神经功能评分(P<0.05)。与假手术组比较,模型组各时间点骨髓及外周血CD 34+EPCs数量及骨髓p-AKT蛋白的表达明显增多(P<0.01,P<0.05);与模型组比较,电针组再灌注后各时间点外周血CD 34+EPCs数量及骨髓p-AKT蛋白的表达亦明显增多(P<0.05,P<0.01),同时电针组再灌注后12、24h骨髓CD 34+EPCs数量相对模型组明显上调(P<0.05,P<0.01)。结论:电针可上调局灶脑缺血/再灌注大鼠骨髓及外周血CD 34+EPCs数量,促进骨髓CD 34+EPCs动员入血,其作用可能与电针进一步激活骨髓磷脂酰肌醇-3-激酶/AKT通路相关。Objective To observe the effect of electroacupuncture （EA） on CD 34^＋ endothelial progenitor cells （EPCs） in bone marrow and peripheral blood and the expression of p-AKT protein in bone marrow in focal cerebral ischemia/reperfusion （CI/R） rats, so as to investigate its mechanism underlying improvement of cerebral ischemia. Methods A total of 108 male SD rats were randomly divided into sham operation （sham） group, model （CI/R） group, and EA group which were further divided into 12, 24, 48 h subgroups （n = 12/group, 6 rats for biochemical analysis and the other 6 rats for Western blot analysis）. CI/R model was established by occlusion of the right middle cerebral artery for 2 hours followed by reperfusion. EA （2 Hz/20 Hz） was applied to ＂Baihui＇（GV 20）, left ＂Hegu＂ （LI 4） and left ＂Taichong＂ （LR 3） acupoints for 30 min, once daily. The neurological deficit scores were evaluated using Longa 5-grade standards. Flow cytometer was used to detect the percentages of CD 34^＋ EPCs in bone marrow and peripheral blood. The expression of p-AKT protein of bone marrow was detected by Western blot. Results In comparison with the CI/R model group, the neurological deficit score were gradually and significantly decreased 48 h after Cl/R in the EA group （P〈0.05）, suggesting an improvement of the neurological function after EA. Compared with the sham group, the percentages of CD 34^＋ EPCs in bone marrow and peripheral blood and the expression level of bone-marrow p-AKT protein were significantly up-regulated in the model group at the three time-points after CI/R （P^0.01, P〈0.05）. Following EA intervention, the percentages of CD 34^＋ EPOs at the three time-points in the peripheral blood, and at time-points of 12 h and 24 h in the bone marrow, and the expression levels of p-AKT protein at the three time-points were significantly further up-regulated in the EA group in comparison with the model group （P〈0.05, P〈0.01 ）. Cenclusion EA can effectively up-regul

关 键 词：电针 局灶脑缺血/再灌注 CD34^+ 内皮祖细胞 磷酸化蛋白激酶B 骨髓 外周血 

分 类 号：R245.97[医药卫生—针灸推拿学]