电针介导eNOS促局灶脑缺血/再灌注大鼠骨髓EPCs动员至外周血的机制研究

The Mechanism of Electroacupuncture Mobilizes Bone Marrow Endothelial Progenitor Cells Via eNOS in Rat with Middle Cerebral Artery Occlusion/Reperfusion

目的探讨内皮型一氧化氮合酶(eNOS)在电针干预下促进局灶脑缺血/再灌注损伤后大鼠骨髓内皮祖细胞(EPCs)动员至外周血的机制。方法用线拴法制备局灶脑缺血/再灌注大鼠模型,选择"合谷"穴及左侧"四关"穴为针刺穴位。60只SD雄性大鼠随机分为假手术组(S组)、模型组(I/R组)、模型+电针组(I/RE组)、模型+电针+eNOS抑制剂(L-NAME)组(I/REL组),再灌注1.5 h后各组又分为24 h、48 h、72 h 3个亚组,每个亚组5只大鼠。采用流式细胞仪检测骨髓及外周血中CD34+EPCs数量,用酶联免疫吸附技术测定外周血eNOS蛋白的表达。结果 I/R大鼠局灶脑缺血/再灌注损伤后24 h、48 h、72 h组骨髓、外周血CD34+EPCs数量及外周血eNOS蛋白表达均明显高于S组(P<0.01),I/RE组各时间点与I/R组比较均显著增高(P<0.01),eNOS抑制剂抑制后各项指标相较于I/RE组显著降低(P<0.01)。结论电针动员骨髓内皮祖细胞至外周血可能与激活eNOS有关。

Objective To explore the mechanism of electroacupuncture（EA）mobilizes bone marrow endothelial progenitor cells（EPCs）via endothelial nitric oxide synthases （eNOS）in rats with middle cerebral artery occlusion/reperfusion（MCAO/R）.MethodsThe focal cerebral ischemia/reperfusion（I/R） model rats were established by filament occlusion of the right middle cerebral artery for 1.5 hours followed by reperfusion.Baihui（GV 20） and Hegu（LI 4）/Taichong（LR 3） were selected as acupuncture points.Sixty healthy male adult SpragueDawley（SD） rats were randomly divided into sham group（S）,model group（I/R）,electroacupuncture group（I/RE）and I/RE plus LNAME（a specific antagonist of eNOS） group（I/REL）,and they were further divided into 24 h,48 h,and 72 h subgroups after reperfusion,5 rats each group.CD34＋ EPCs cells were measured by flow cytometer in bone marrow and peripheral blood.Enzymelinked immunosorbent assay（ELISA）was used to detect the expression of eNOS protein in peripheral blood.ResultsThe number of CD34＋EPCs and the expression of eNOS protein in the peripheral blood were significantly higher in I/R 24 h,48 h,and 72 h subgroups than those in S group（P〈0.01）.Compared with I/R 24 h,48 h,and 72 h subgroups,the number of CD34＋EPCs and the expression of eNOS increased in I/RE group.Compared with I/RE group,the indicators were significantly lower in I/REL group（P〈0.01）.ConclusionEA can mobilize bone marrow EPCs,which may be associated with activation of eNOS.