类缺血再灌注损伤对体外培养神经干细胞活性和胞内钙浓度及增殖的影响

目的观察类缺血再灌注损伤对体外培养的胚胎小鼠纹状体神经干细胞活性、胞内Ca2+浓度以及增殖的影响。方法将体外培养的小鼠神经干细胞分为类缺血再灌注组、药物预处理组(1μmol/L氟桂利嗪)和对照组。前两组进行类缺血处理并分别于再灌注0、30 min、1、2、3 h检测细胞内Ca2+浓度,于03、0 min、12、、31、2、24、36、486、0 h检测细胞活性,对照组在相同时间点检测细胞活性,比较3组间细胞内Ca2+浓度及细胞活性的差异。结果类缺血再灌注后神经干细胞内游离Ca2+浓度显著升高,在相同时间点药物预处理组胞内Ca2+浓度均低于类缺血再灌注组,其中0、30 min1、h时有显著性差异(P<0.05);类缺血再灌注后细胞活性降低,在相同时间点药物预处理组细胞活性均高于类缺血再灌注组,其中03、0 min、1、2 h时差异显著(P<0.05)。再灌注后12~60 h类缺血再灌注组和药物预处理组的细胞增殖率明显高于对照组(P<0.01)。结论类缺血再灌注可以导致胞内Ca2+浓度升高及神经干细胞活性的降低,氟桂利嗪预处理可抑制胞内Ca2+的升高,减轻神经干细胞的损伤;另外类缺血再灌注后存活的神经干细胞的增殖能力增强。

Effects of ischemia-reperfusion-like condition on intracellular Ca2+ activity and proliferation of neural stem cells cultured in vitro

Objective To study the effects of ischemia-reperfusion-like condition on intracellular Ca~(2+),activity and proliferation of neural stem cells of fetal mouse corpus striatum cultured in vitro.Methods Neural stem cells cultured in vitro were divided into three groups:ischemia-reperfusion group,pretreatment(flunatizine 1 μmol/L) group and control group.The changes of intracellular Ca~(2+) were observed at 0,30 min,1,2,3 h after ischemia-reperfusion and the cell activity were tested at 0,30 min,1,2,3,12,24,36,48,60 h after ischemia-reperfusion.The differences of intracellular Ca~(2+) and cell activity among three groups were compared.(Results)The intracellular Ca~(2+) increased after ischemia-reperfusion.The intracellular Ca~(2+) of pretreatment group was lower than that of ischemia-reperfusion group at the same time point,and the difference was obvious at 0,30 min,1 h(P<0.05).The cell activity decreased after ischemia-reperfusion.The cell activity of pretreatment group was higher than that of ischemia-reperfusion group at the same time point,and the difference was obvious at 0,30 min,1,2 h(P<0.05).Cells of ischemia-reperfusion group and pretreatment group proliferated faster than control obviously from 12 h to 60 h after ischemia-reperfusion(P<0.01).Conclusions Ischemia-reperfusion led to the increase in intracellular Ca~(2+) and the decrease in cell activity.Flunarizine can prevent the increase in intracellular Ca~(2+) and reduce the cell damage.In addition,ischemia-reperfusion-like condition may improve the proliferating ability of the surviving neural stem cells.