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

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(P0.05)。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(P0.01,P0.05);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(P0.05,P0.01),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(P0.05,P0.01)。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。

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

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 into12,24,48hsubgroups(n=12/group,6rats for biochemical analysis and the other 6rats for Western blot analysis).CI/R model was established by occlusion of the right middle cerebral artery for 2hours followed by reperfusion.EA(2Hz/20Hz)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 hafter CI/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+EPCs at the three time-points in the peripheral blood,and at time-points of 12 hand 24hin 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).Conclusion EA can effectively up-regulate the percentages of CD 34+EPCs in the bone marrow and peripheral blood,and increase p-AKT protein expression in the bone marrow in CI/R rats,which may contribute to its effect in improving neurological function.