摘要
目的从细胞凋亡及其信号转导通路角度,探讨盐酸法舒地尔对脑缺血再灌注损伤大鼠的脑保护作用。方法雄性健康SD大鼠72只,随机分成3组:假手术组(S组,n=24)、缺血再灌注组(IR组,n=24)、盐酸法舒地尔组(F组,n=24),三组按再灌注时间再分为3个亚组,即缺血10 min后分别再灌注3、12、24 h,每个亚组动物均为8只。采用四血管闭塞法(4-vessel-occlusion,4VO)制造大鼠全脑缺血模型,应用免疫组化SP法动态观察不同时间点海马CA1区细胞色素C(cytC)表达的变化;电镜和光镜观察再灌注24 h亚组海马CA1区神经细胞病理形态和线粒体超微结构的改变。结果①大鼠脑缺血再灌注后海马CA1区cytC在再灌注3 h即有明显表达,于再灌注12 h达高峰,以后表达逐渐降低;②与S组比较,IR组和F组各时间点海马CA1区的cytC蛋白表达明显升高(P<0.01);再灌注24 h亚组海马CA1区神经细胞病理形态和线粒体形态结构受损明显;③与IR组比较,F组各时间点海马CA1区的cytC蛋白表达明显降低(P<0.05或P<0.01);再灌注24 h亚组海马CA1区神经细胞病理形态和线粒体形态结构均有不同程度的改善。结论盐酸法舒地尔对CytC介导的线粒体凋亡通路有干预作用,通过抑制cytC的释放和激活,稳定线粒体膜,保护线粒体的形态功能,从而减少细胞凋亡的发生,发挥脑保护作用。
Objective To evaluate the effects of hydrocloric fasudil on acute focal cerebral ischemia/reperfusion（IR） injury in rats.Methods Male SD rats were randomly divided into three groups： group S（sham operation group,n=24）,group IR（ischemia/reperfusion,n=24）,group F（Fasudil treated group,n=24）.In each group the rats were treated with 10 min ischemia and 3,12,24h cerebral reperfusion,respectively with 8 rats in each subgroup.The model of focal cerebral ischemia reperfusion injury was established by using Pulsinelli＇s method.Immunohistochemical SP method was performed to examine the release of cytochrome C（cytC）at different time points after reperfusion;The pathological changes of nerve cells and mitochondria ultrastructure at hippocampus CA1 region were observed at 24h after reperfusion by light microscope and electron microscope.Results The activation of cytC began to increase at 3h after reperfusion,reached to the peak at 12h and then gradually decreased.Compared with group S,the cytC levels in group IR and F were increased significantly（P0.01）.The pathological changes of nerve cells and mitochondria ultrastructure in hippocampus CA1 region were exacerbated at 24h after reperfusion.Compared with IR group,the cytC levels in group P were decreased significantly（P0.05 or P0.01）.The pathological changes of nerve cells and mitochondria ultrastructure in hippocampus CA1 region were improved at 24h after reperfusion.Conclusion Hydrocloric Fasudil intervenes in mitochondria-mediated and CytC-dependent apoptosis pathway.Hydrocloric Fasudil attenuates apoptosis,and protects nerve cells by maintaining mitochondrial basic form,stabilizing mitochondrial membrane and inhibiting the release of cytC.
出处
《同济大学学报（医学版）》
CAS
2012年第6期36-40,共5页
Journal of Tongji University(Medical Science)
关键词
盐酸法舒地尔
脑缺血再灌注损伤
线粒体
细胞色素C
hydrocloric Fasudil
cerebral ischem/reperfusion injury
mitochondria
cytochrome C
