论文部分内容阅读
目的:研究CASPASE-9在英国皇家外科学院(RCS)变性大鼠视网膜的时空表达状况与其视网膜电图 (ERG)以及感光细胞凋亡的关系,以探讨CASPASE-9在感光细胞凋亡中的作用。方法:不同日龄RCS变性大鼠,检查 ERG后处死取视网膜行CASPASE-9的免疫组化、荧光活力分析、WESTERN BLOTTING及凋亡TUNEL检测。结果:15及20 D 变性大鼠ERG B波潜伏期分别为(58．60±3．42)MS、(56．45±1．08)MS,振幅分别为(109．07±14．50)MV、(109．00±7．35) MV,两组间无显著性差异,25和30D组接近熄灭型。TUNEL阳性细胞只在变性大鼠的外颗粒层表达,25D最明显。 25D后的变性大鼠感光细胞内节可见明显CASPASE-9阳性表达,其余各组未见表达。20 D CASPASE-9表现出最大活力[(10．98±0．13)NMOL·G-1·MIN-1]。WESTERN BLOTTING分析显示CASPASE-9裂解条带在20 D时最明显,15 D组和对照组不表达。结论:CASPASE-9的表达与视网膜感光细胞凋亡、视功能丧失存在时空一致性。
AIM: To investigate the temporal and spatial expression of caspase-9 in the retina of the RCS rat model and its relationship with ERG and photoreceptor cell apoptosis in order to investigate the role of caspase-9 in photoreceptor cell apoptosis . Methods: The RCS-denatured rats of different ages were examined by immunohistochemistry, fluorescence assay, WESTERN BLOTTING and apoptotic TUNEL assay of CASPASE-9 after ERG. Results: The latent periods of ERG B wave in 15 and 20 D degeneration rats were (58.60 ± 3.42) MS and (56.45 ± 1.08) MS respectively, and the amplitudes were (109.07 ± 14.50) MV , (109.00 ± 7.35) MV, no significant difference between the two groups, 25 and 30D group near quenching type. TUNEL-positive cells were only expressed in the outer layer of denatured rats, 25D most obvious. After 25 days, the expression of CASPASE-9 in the degenerated rat photoreceptor cells was obvious, and the rest of the groups showed no expression. 20 D CASPASE-9 showed the highest activity [(10.98 ± 0.13) NMOL · G-1 · MIN-1]. WESTERN BLOTTING analysis showed that CASPASE-9 cleavage band at 20 D most obvious, 15 D group and the control group does not express. CONCLUSION: There is spatiotemporal consistency between CASPASE-9 expression and retinal photoreceptor apoptosis and loss of visual function.