电针对局灶性脑缺血/再灌注大鼠外周血和骨髓内皮祖细胞的作用

目的:探讨电针对局灶性脑缺血/再灌注大鼠外周血及骨髓中与血管再生相关的因子及细胞的影响。方法:54只SD大鼠随机分为正常组、模型组和电针组,采用线栓法制备局灶性脑缺血/再灌注模型,并按局灶性脑缺血2h再灌注后的观察时间点,将模型组和电针组分为1、2、3、7d各4个亚组,每个时间点6只大鼠。电针刺激大鼠双侧"合谷"穴,刺激时间15min,每日1次。采用流式细胞术检测外周血内皮祖细胞(EPCs)、趋化因子受体4+(CXCR 4+)细胞和骨髓EPCs占单核细胞的百分比,酶联免疫法检测血清中基质细胞衍生因子-1α(SDF-1α)蛋白浓度。结果:局灶性脑缺血/再灌注大鼠1d模型组和电针组较正常组外周血EPCs和CXCR 4+细胞数量均显著增高(P<0.01),2d电针组高于模型组(P<0.05);2d电针组骨髓EPCs数量显著高于正常组和模型组(P<0.05,P<0.01);模型组和电针组血清SDF-1α蛋白浓度第1天均增至最高,较正常组差异具有统计学意义(P<0.05,P<0.01),且第1、2天电针组显著高于模型组(P<0.01)。结论:电针能促进局灶性脑缺血/再灌注大鼠外周血EPCs、CXCR 4+细胞和骨髓EPCs数量的增加,该作用可能与SDF-1α表达上调相关。

Effects of electroacupuncture on endothelial progenitor cells in peripheral blood and bone marrow in local cerebral ischemia/reperfusion rats

Objective To observe the influence of electroacupuncture(EA) of "Hegu"(LI 4) on blood stromal cell-derived factor 1α(SDF-1α),CXC chemokine receptor 4+(CXCR 4+) positive cells and endothelial progenitor cells(EPCs) and bone marrow(BM) EPCs levels in local cerebral ischemia/reperfusion injury(CI/IR) rats,so as to study its mechanisms underl-ying improvement of cerebral ischemia.Methods A total of 54 SD rats were randomly divided into normal control,model,and EA groups.The latter two groups were further divided into 1 d,2 d,3 d and 7 d subgroups(4 time-points),respectively.CI/RI model was established by occlusion of the middle cerebral artery.EA(40 Hz/60 Hz,1-2 mA) was applied to bilateral "Hegu"(LI 4) for 15 min,once daily,1,2,3 and 7 days,respectively.Flow cytometer was used to detect the counts of EPCs and CXCR 4+ cells in the peripheral blood,and BM EPCs.Serum SDF-1α was detected by enzyme linked immunosorbent assay(ELISA).Results Compared with the normal control group,the percentages of blood EPCs and CXCR 4+ cells and BM EPCs,and serum SDF-1α content on the 1st day,and the percentage of blood CXCR 4+ cells on the 2nd day were increased significantly in the model group(P<0.01,P<0.05).In comparison with the model group,percentages of blood EPCs and CXCR 4+ cells on the 1st day of EA group were reduced significantly(P<0.01),but the percentage of blood EPCs on the 2nd day,those of blood CXCR 4+ cells on the 2nd day and 3rd day,serum SDF-1α content on the 2nd day,and the percentage of BM EPCs on the 2nd day in the EA group were all up-regulated significantly(P<0.05,P<0.01).No significant differences were found among the 3 groups in blood EPCs percentages on the 3rd day and 7th day,in blood SDF-1α content and BM EPCs percentage on the 7th day(P>0.05).Conclusion Generally,2 days’ EA of LI 4 can up-regulate the percentages of blood and BM EPCs and blood CXCR 4+ cells as well as blood SDF-1α protein content in CI/RI rats,which may contribute to its effect in improving cerebral ischemia.