天津医药

天津医药 ›› 2016, Vol. 44 ›› Issue (6): 694-699.doi: 10.11958/20160042

• 实验研究 • 上一篇    下一篇

血管内皮祖细胞-血管再生在缺血再灌注损伤中的作用探讨

张智申， 董贝贝， 谢克亮， 于泳浩   

  1. 天津医科大学总医院麻醉科， 天津市麻醉学研究所
  • 收稿日期:2016-01-29 修回日期:2016-03-14 出版日期:2016-06-15 发布日期:2016-07-04
  • 基金资助:
    国家自然科学基金资助项目 （815010139）

The potential effects of EPCs-angiogenesis on ischemia-reperfusion injury

ZHANG Zhishen, DONG Beibei, XIE Keliang, YU Yonghao   

  1. Anesthesiology Department of General Hospital of Tianjin Medical University, Anesthesiology Research Institute of Tianjin Medical University, Tianjin 300052, China
  • Received:2016-01-29 Revised:2016-03-14 Published:2016-06-15 Online:2016-07-04

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摘要： 摘要： 目的 探讨血管内皮祖细胞 （EPCs） -血管再生途径在血红素结合蛋白 （HPX） 改善大鼠脑缺血再灌注 （I/R）后认知功能障碍中的作用。方法 SD 雄性大鼠依处理方式不同分为假手术（Sham）组 31 只、 局灶性 I/R（MCAO，0.9%生理盐水 10 μL）组 30 只、 MCAO+叠氮钠（Vehicle， 0.1% Vehicle 10 μL）组 30 只、 MCAO+ HPX（1.86 g/L 的 HPX10 μL）组 30 只。采用改良的神经功能缺损评分（mNSS）评价大鼠 I/R 后神经功能缺损程度，Morris 水迷宫（MWM）实验检测大鼠 I/R 后的学习记忆能力改变； 流式细胞技术 （FCM）结合 CD34/CD133 双抗体标记技术检测大鼠 I/R 后外周血中 EPCs 数量变化； 免疫组织化学技术结合 CD31/vWF 免疫荧光显色技术检测大鼠 I/R 后缺血半暗带皮质区新生血管数量。 结果 与 Sham 组相比， MCAO 组大鼠 I/R 后 mNSS 升高， 逃避潜伏期延长， 目标象限时间百分比减少， 外周血中 EPCs 数量增加， 缺血半暗带皮质区血管数量增加（均 P < 0.05）； MCAO+Vehicle 组大鼠与 MCAO 组相比， I/R 后 mNSS、 逃避潜伏期、 目标象限时间百分比、 外周血中 EPCs 数量和缺血半暗带皮质区血管数量均无明显变化 （均 P > 0.05）； MCAO+HPX 组大鼠与 MCAO 组、 MCAO+Vehicle 组相比， I/R 后 mNSS 显著降低， 逃避潜伏期显著缩短， 在目标象限时间百分比明显增加， 外周血中 EPCs 计数显著增加， 缺血半暗带皮质区血管数量明显增加（均 P <0.05）。结论 EPCs-血管再生机制在 HPX 改善大鼠局灶性 I/R 后认知功能障碍中发挥积极的作用。

关键词: 脑缺血, 再灌注损伤, 血红素结合蛋白, 血管内皮祖细胞, 血管再生

Abstract: Abstract：Objective To explore the potential effects of endothelial progenitor cells (EPCs)- angiogenesis on mechanism of alleviating cognitive dysfunction in rats subjected to cerebral ischemia-reperfusion (I/R) injury. Methods Atotal of 121 male Sprague–Dawley (SD) rats were randomly divided into four groups: Sham group (n=31), focal I/R(MCAO,0.9% saline 10 μL, n=30) group, MCAO+Vehicle (sodium azide, 0.1% Vehicle 10 μL, n=30) group and MCAO+HPX (1.86 g/L HPX 10 μL, n=30) group. The modified neurological severity scores (mNSS) was carried out to determine neurological function deficit after I/R. Morris water maze (MWM) was carried out to assess learning and memory abilities after I/R. The circulating EPCs after I/R were counted by flowcytometry (FCM) combined with double- immunofluorescence staining of CD34 and CD133. Angiogenesis in rat penumbra cortex after I/R was assessed by immunohistochemical technique combined with immunofluorescent chromogenic detection of CD31 and vWF. Results Compared with sham group, the mNSS scores, the escape latency and the circulating EPCs count were increased after I/R, the time percentage spent in target quadrant was reduced, and the new vessel density in penumbra cortex was increased after I/R in MCAO group (P < 0.05 respectively).There were no significant differences in mNSS score, the escape latency, the time percentage spent in target quadrant, the circulating EPCs count and the new vessel density in penumbra cortex between MCAO group and MCAO+Vehicle group (P >0.05). The mNSS score and the escape latency were significantly decreased, the circulating EPCs count and new vessel density in penumbra cortex were significantly increased after I/R in MCAO+HPX group compared with those of MCAO+Vehicle and MCAO group (P < 0.05). Conclusion EPCs-angiogenesis signaling plays positive effects on HPX alleviating cognitive dysfunction in rats subjected to focal cerebral ischemia reperfusion injury.

Key words: brain ischemia, reperfusion injury, hemopexin, endothelial progenitor cells, angiogenesis