RCS变性大鼠感光细胞凋亡及其视网膜电图与CASPASE-9时空表达的关系研究  

Relation between the time - spatiotemporal expression of caspase-9 and photoreceptor apoptosis as well as electroretinography in RCS rats

作　　者：朱晓波[1] 罗燕[1] 谢琳[1] 孟晶[1] 丁小燕[1] 李加青[1] 林少芬[1] 唐仕波[1] 

机构地区：[1]中山大学中山眼科中心,广东广州510060

出　　处：《中国病理生理杂志》2005年第12期2438-2442,共5页Chinese Journal of Pathophysiology

基　　金：国家自然科学基金资助项目(NO.30200307);广东省自然科学基金重点项目(NO.021827);广东省医学科学技术研究基金资助项目(NO.B2002045)

摘　　要：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 relation between the time - spatiotemporal expression of caspase - 9 and photoreceptor apoptosis as well as electroretinography （ERG） in Royal College of Surgeons （RCS） rats.METHODS： Twelve non- pigmented degenerative RCS rats at age of 15, 20, 25, 30 days were sacrificed after ERG examination. The caspase - 9 expression was analyzed with immunohistochenfical staining, fluoro - metric activity assay and Western blotting. Apoptotic retinal neurons were detected by terminal deoxynucleotidyl transferase- mediated dUTP nick- end labeling （TUNEL） technique. RESULTS： The latency and amplitude of b - wave were （58.60 ± 3.42） ms and （ 109.07 ± 14.50） mV, （56.45 ± 1.08） ms and （ 109.00 ± 7.35） mV, respectively, in 15 and 20 days old degenerative RCS rats. The ERG resembled fiat form in the degenerative RCS rats at age of 25 and 30 days. TUNEL- positive cells were located only in the outer nuclear layer, mostly prevailed in 25 - day - old group. Caspase - 9 protein expressed in the rat retina started to be found immunohistochemically in 25 - day - old and was most predominant in the inner segment. Huorometric activity assay showed that caspase- 9 activity reached its peak, （10.98 ± 0.13 ） nmol·g^-1, min^-1 at 20th day. Western blot assay revealed the small cleavage fragmentation at 20th day. No cleavage fragmentation was revealed in the 15 - day - old degenerative RCS rats and all other control rats. CONCLUSION： The activation and time- spatiotemporal expression of caspase - 9 are coincidence with photoreceptor apoptosis as well as the loss of vision function in RCS degenerative rats, which implicates the important role of caspase - 9 in photoreceptor apoptosis.

关 键 词：视网膜变性 细胞凋亡 半胱氨酸天冬氨酸蛋白酶-9 

分 类 号：R927[医药卫生—药学]