大鼠局灶脑缺血/再灌注后骨髓内皮祖细胞动员轨迹的追踪  

Tracing the trajectory of the mobilization of bone-marrow EPCs in rats after middle cerebral artery occlusion/reperfusion

作　　者：朱艳含 罗勇[1] 胥虹贝 王盼欣 

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3A期229-235,共7页Chinese Journal of Histochemistry and Cytochemistry

基　　金：教育部"高等学校博士学科点专项科研基金"联合资助(20095503110001);重庆市卫生局中医药科研重点资助(ZY20131027);重庆市卫生局中医药科研项目资助(渝中医2005-B-24)

摘　　要：目的观察大鼠局灶脑缺血/再灌注后骨髓内皮祖细胞(EPCs)能否动员至外周血进而归巢到缺血脑区。方法线栓法制备局灶脑缺血/再灌注模型。130只SD雄性大鼠完全随机分为模型+生理盐水组、模型+DIL-acLDL组。荧光共聚焦技术观察1d后骨髓腔内细胞摄取DIL-acLDL的情况;1d、2d、3d、7d观察骨髓、外周血CD34+DIL-acLDL+双阳性细胞及缺血大脑皮质区DIL-acLDL+阳性细胞情况;免疫组织化学法比较各组大鼠大脑缺血侧及未缺血侧皮质区CD34+血管表达;荧光定量PCR法比较各组大鼠大脑缺血侧及未缺血侧皮质区CD34mRNA表达。结果生理盐水组荧光共聚焦结果为阴性,DIL-acLDL组1d后骨髓腔内观察到红色标记的DIL-acLDL+阳性细胞,并分别于1d、2d、3d、7d后在骨髓及外周血中观察到CD34+DIL-acLDL+双阳性细胞,在缺血大脑皮质区未检测到DIL-acLD L+阳性细胞,但3d、7d缺血大脑皮质侧CD34+血管及CD34m RNA的表达明显高于未缺血侧(P<0.01,P<0.05),且表达量均随时间的推移而增多(P<0.01)。结论DIL-acLDL可标记活体大鼠骨髓腔内细胞,且骨髓腔内EPCs可动员至外周血并促大脑缺血皮质区血管再生。Objective To observe the mobilization of bone-marrow endothelial progenitor cells(EPCs)to peripheral blood and homing to the ischemic brain regions of rats after focal cerebral ischemia/reperfusion.Methods The rats received occlusion of the right middle cerebral artery for 1.5hours followed by reperfusion.A total of 130 healthy male adult Sprague-Dawley(SD)rats were randomly divided into I/R+NS group and I/R+DIL-acLDL group.Immunofluorescence was used to test bone marrow DIL-acLDL+cells,bone marrow and peripheral blood CD34+DIL-acLDL+ cells,and DIL-acLDL+ cells in ischemic brain regions.Immunohistochemical staining was used to detect CD34+microvessels.The expression of CD34 mRNA was tested by fluorescence quantitative PCR.Results The results of the I/R+NS group were negative.DIL-acLDL+cells were found in bone marrow 1dafter intra-bone marrow injection,and CD34+DIL-acLDL+cells were found 1d,2d,3dand 7dafter intra-bone marrow injection respectively in bone marrow and peripheral blood.DIL-acLDL+cells were not found in ischemic brain regions after intra-bone marrow injection.Compared with the non-ishemic cerebral cortex,the expression of CD34+vessels and CD34 mRNA significantly increased in the ischemic cerebral cortex(P<0.01,P<0.05),and the expression quantity was increasing with the passage of time(P<0.01).Conclusion In vivo,bone-marrow cells can be tagged by DIL-acLDL in rats after focal cerebral ischemia/reperfusion.Bone-marrow CD34+DILacLDL+ EPCs can also be tested in peripheral blood,demonstrating the moving trajectory of CD34+DILacLDL+ EPCs directly.

关 键 词：局灶脑缺血/再灌注 内皮祖细胞 DIL-acLDL CD34 

分 类 号：R743.33[医药卫生—神经病学与精神病学]