VEGF提高MCAO/R小鼠的EPCs表达及促进脑梗死的康复

目的：观察局灶性脑缺血再灌注模型（MCAO/R）小鼠外周循环中血管内皮祖细胞（EPCs）的数量变化,并应用血管内皮生长因子（VEGF）动员小鼠自体骨髓中的EPCs,达到治疗脑梗死的目的。方法：以MCAO/R小鼠为研究对象,通过腹腔注射VEGF动员体内的EPCs,（每日一次,持续一周）,在动员过程中的第1d、4d、7d、14d从内眦静脉采血,使用流式细胞仪检测MCAO/R组及MCAO/R＋VEGF组小鼠外周血中EPCs的数量;然后断头取脑,用免疫组化法检测新生血管的密度,并对脑组织TTC染色后计算并比较小鼠脑梗死体积的变化。结果：MCAO/R＋VEGF组在动员过程中,外周血中的EPCs在给药第1d开始增加,第4d达到高峰并持续到第7d,第14d高峰持续存在;并且在这四个时间点上,MCAO/R＋VEGF组与其它两组比较均具有统计学意义（P〈0.01）。新生血管密度：MCAO/R＋VEGF组1d开始有新生血管生成,4d血管新生明显,14d达高峰,且各时间点MCAO/R＋VEGF组血管新生数量明显多于MCAO/R组。TTC染色后通过图像分析系统计算脑梗死体积：MCAO/R＋VEGF组在给药第14d较第1d、4d、7d梗死灶体积明显缩小（P〈0.01）,且MCAO/R＋VEGF组在给药第4d,7d、14d的梗死灶体积较同时间点的MCAO/R组有显著缩小（P〈0.01）。结论：在急性脑缺血/再灌注损伤的情况下,应用VEGF能显著动员小鼠骨髓中的EPCs,增强出生后的血管新生,达到缩小梗死灶,治疗脑梗死的目的。

Experimental study on the treatment of brain infarction by VEGF through mobilizing endothelial progenitor cells

Objective：To observe the change of number of endothelial progenitor cells（EPCs） in peripheral circulation after acute MCAO/R and evaluate treatment of mouse brain infarction after acute MCAO/R by VEGF through mobilizing bone marrow-derived EPCs.Methods： We injected VEGF intraperitoneally to mobilize bone marrow-derived EPCs once a day for one week.At day 1,4,7,14 during mobilization,blood samples were taken from angular vein.Then we detected the number of EPCs in peripheral circulation in MCAO/R group and MCAO/R＋VEGF group by flow cytometry.Finally,mice were decapitated and brains sliced and stained with TTC.Infarct volumes were calculated and compared among groups.Results：EPCs in MCAO/R＋VEGF group began to increase at day 1 after treatment,and peaked at day 4 and sustained to day 7 and 14;while number of EPCs in operatedmobilization group had significant difference between groups（p〈0.01）,blood vessel density： MCAO/R＋VEGF group ld began to angiogenesis,4d angiogenesis significantly,14d reached the peak,and each time point MCAO/R＋VEGF group was higher than the number of angiogenesis in MCAO/R group.After TTC staining,infarct volume was calculated using specific image analyzing system.Infarct volume decreased significantly in operated-mobilization group at day 14 compared with that at day 1（p〈0.01） and as is the case at day 14 compared with that at day 4,day 7（p 〈0.01）.It showed significant difference when infarct volumes were compared between MCAO/R＋VEGF group and MCAO/R group at day 14 and day 4,day 7 respectively（p〈0.01）,infarct volume in MCAO/R＋VEGF group was smaller than that in MCAO/R group.Conculsion： In the state of acute MCAO/R,VEGF can mobilize EPCs in mouse bone barrow.The mobilized EPCs can increase angiogenesis to decrease infarction area and treat brain infarction.