目的:研究蒙药珍宝丸对Fas、FasL及p53蛋白在家兔视网膜缺血再灌注损伤中的表达影响,探讨珍宝丸对家兔视网膜缺血再灌注损伤的保护作用及其相关机制。方法:通过升高眼内压法造成视网膜缺血再灌注损伤模型,采用免疫组织化学链酶卵白素一生物素复合体法和计算机图像分析系统对95只家兔视网膜中Fas、Fasl和P53蛋白表达进行测定。结果:1.组织病理学改变:6~48 h水肿逐渐加重,神经节细胞和内核层细胞数量减少,内外核层密度降低、排列紊乱。72 h视网膜明显变薄,水肿减轻,神经节细胞数量稀少,内核层明显变薄。2.相关因子表达情况:Fas在正常视网膜组织中几乎不表达,在缺血再灌注6 h开始表达,24 h达到高峰,48 h开始下降,延续至缺血再灌注后72 h仍有表达;FasL在正常视网膜组织中低表达,于缺血再灌注12 h表达升高,24 h达到高峰,48 h开始下降;再灌注72 h表达较24 h降低,但仍维持高表达。P53在正常视网膜组织中无表达,缺血再灌注后6 h开始表达,24 h达高峰,48 h仍持续强表达,72 h已明显下降。珍宝丸治疗组各观察指标表达变化规律与缺血再灌注组基本一致,但均较缺血再灌注组明显下降。其中Fas表达在6 h、12 h、24 h、48 h组较缺血再灌注组显著下降(P<0.05);FasL表达在12 h、24 h、48 h组较缺血再灌注组明显下降(P<0.05);P53在6 h、12 h、24 h、48 h组表达明显低于缺血再灌注组(P<0.05)。结论:视网膜缺血再灌注损伤导致视网膜组织中Fas/Fasl和P53蛋白表达的增高。珍宝丸可以抑制其增高,可用于临床上视网膜缺血再灌注损伤的治疗。

The study was aimed to explore basic protective effects of retinal ischemia-reperfusion injury and effects on the expression of Fas,Fasl,P53 and apoptosis occured in retinal ischemia-reperfusion injury of rabbit model.Ninety-five healthy rabbits without eye disorders were selected.They were randomly divided into four groups,which are normal control group,ischemia-reperfusion group,Zhen-Bao Pills group,and VE group.According to time after reperfusion,ischemia-reperfusion group,Zhen-Bao Pills group and VE group were further divided in to 1 h,6 h,12 h,24 h,48 h and 72 h subgroup.Rabbit model of experimental retinal ischemia-reperfusion injury was made by increasing intraocular pressure.Then,the sample was processed with HE staining and immunohistochemical(SABC) staining.Histological and ultrastructural changes in retina were observed.Results of SABC staining were analyzed by compute picture analytic system.The result showed no Fas positive cells in the normal retina.Fas expression was gradually increased at 6 h.It reached a peak at 24 h.And then,it decreased at 48 h.Similarly,Fasl expression was at peak from 24 h to 48 h in GCL and INL of retina.And the peak of P53 expression was at 24 h.Then it was decreased gradually at 72 h.Zhen-Bao Pills administered before reperfusion ameliorated the retinal tissue damage.It also diminished Fas,Fasl and P53 expression in retinal ischemia-reperfusion remarkably.Fas expression was obviously decreased at 6 h,12 h,24 h,and 48 h in treatment group.Zhen-Bao Pills decreased Fasl expression at 12 h,24 h,48 h.P53 expression was decreased at 6 h,12 h,24 h,48 h.It is concluded that retinal ischemia-reperfusion injury can cause increasing of Fas\Fasl and P53 protein in retina.Zhen-Bao Pills can inhibit the increasing of Fas\Fasl and P53 protein in retinal ischemia-reperfusion injury.Zhen-Bao Pills have protective effects against retinal ischemia-reperfusion injury.It can be used in the clinical treatment of ischemia-reperfusion injury.

内蒙古医学院内蒙古自然科学基金资助项目(2009MS1159):蒙药额尔顿-乌日勒对视网膜缺血再灌注损伤的保护机理研究,负责人:乌仁图雅