AMD3100通过阻断SDF-1／CXCR4通路减少脑慢性低灌注模型大鼠血管新生  

AMD3100 decreasing angiogenesis in rat models of cerebral chronic hypoperfusion by blocking SDF-1/CXCR4 pathway

作　　者：王凌雁[1] 张恒[1] 郭少雷[2] 黄正松[2] 

机构地区：[1]中山大学附属第一医院神经外科ICU,广州510080 [2]中山大学附属第一医院神经外科,广州510080

出　　处：《中华神经医学杂志》2015年第8期764-769,共6页Chinese Journal of Neuromedicine

基　　金：国家自然科学基金面上项目（81171102）

摘　　要：目的探讨AMD3100对脑慢性低灌注模型大鼠血管新生的影响及其作用机制。方法36只SD大鼠按随机数字表法分为正常对照组、脑慢性低灌注模型＋AMD3100组和脑慢性低灌注模型＋生理盐水组，每组12只。后两组将大鼠一侧颈外静脉和颈总动脉端侧吻合，同时夹闭双侧颈外动脉及对侧横窦引流静脉建立脑慢性低灌注模型，并分别于手术当天开始腹腔注射AMD31005mg／（kg·d1或同等体积生理盐水，连续30d。然后处死各组大鼠并取左侧大脑中动脉供血区皮层组织，根据CD34免疫组化染色结果计算微血管密度（MVD），免疫荧光染色双标法标记CD45和CXCR4共表达细胞，Westernblotting检测基质金属蛋白酶-9（MMP-9）蛋白表达情况。结果与正常对照组MVD（61．82±19．83／mm^2）及CD45、CXCR4双阳性细胞数（3．47±2．63）及MMP．9蛋白表达量（0．009±0．0031比较．脑慢性低灌注模型＋AMD3100组MVD（89．50±23．61／mm^2）及CD45、CXCR4双阳性细胞数（7．58±4．49）及MMP-9蛋白表达量（0．151±0．058）以及脑慢性低灌注模型＋生理盐水组MVD（121．70±31．12／mm^2）及CD45、CXCR4双阳性细胞数（12．33±6．11）及MMP-9蛋白表达量（0．325±0．068）均明显增高，差异均有统计学意义（P〈0．05）；脑慢性低灌注模型＋AMD3100组MVD及CD45、CXCR4双阳性细胞数及MMP-9蛋白表达量均明显低于脑慢性低灌注模型＋生理盐水组，差异有统计学意义（P〈0．05）。结论对脑慢性低灌注模型大鼠应用AMD3100可以抑制血管新生，其机制可能与阻断基质细胞衍生因子-1（SDF-1）／CXCR4通路从而影响表达CXCR4的CD45阳性细胞向脑内浸润有关。Objective To observe the effect of ADM3100 on angiogenesis in rat models of cerebral chronic hypoperfusion （CCH） and its mechanism. Methods Thirty-six healthy female SD rats, weighting 200-250 g, were randomly divided in normal control group, CCH＋AMD3100 group and CCH＋saline group （n=12）; rats in the CCH＋AMD3100 group and CCH＋saline group underwent end-to-side anastomosis between the left distal external jugular vein （EJV） and the ipsilateral common carotid artery, followed by ligation of the right vein draining the transverse sinus and bilateral external carotid arteries, and then, they were received intraperitoneal injection of 5 mg/ （kg. d） ADM3100 and physiological saline, respectively, for a consecutive 30 d. Rats were then scarified; microvessel density （MVD） was assessed based on immunohistochemistry of CD34, numbers of CXCR4 and CD45 double positive cells were confirmed by double immunofluorescence, and matrix metalloproteinases-9 （MMP-9） was evaluated by Western blotting. Results MVD was 61.82±19.83/mm^2, 89.50±23.61/mm^2 and 121.70 ±31.12/mm^2 in the normal control group, CCH＋AMD3100 group and CCH＋saline group, respectively; numbers of CXCR4 and CD45 double positive cells were 3.47±2.63, 7.58±4.49 and 12.33±6.11 per high power field （x200） in normal control group, CCH＋AMD3100 group and CCH＋saline group, respectively; significant differences between groups were noted （P〈0.05）. As compared with the expression of MMP-9 in the normal control group, that in the CCH＋AMD3100 group and CCH＋saline group was significantly increased （0.009±0.003, 0.151 ±0.058 and 0.325 ±0.068, P〈0.05）. As compared with CCH＋saline group, CCH＋AMD3100 group had significantly smaller MVD and numbers of CXCR4 and CD45 double positive cells and lower MMP-9 level （P〈0.05）. Conclusion AMD3100 will decrease angiogenesis in rat models of cerebral chronic hypoperfusion, which may be associated with inhibiting CXCR4＋ CD45＋ cells infiltration into cerebral tissu

关 键 词：脑慢性低灌注 基质细胞衍生因子-1 CXCR4 血管新生 AMD3100 

分 类 号：R743.4[医药卫生—神经病学与精神病学] R-332[医药卫生—临床医学]