中华眼视光学与视觉科学杂志
中華眼視光學與視覺科學雜誌
중화안시광학여시각과학잡지
CHINESE JOURNAL OF OPTOMETRY OPHTHALMOLOGY AND VISUAL SCIENCE
2015年
6期
350-355
,共6页
视网膜变性%N-亚硝基-N-甲基脲%显微镜检查,电子%视网膜电描记术%小鼠
視網膜變性%N-亞硝基-N-甲基脲%顯微鏡檢查,電子%視網膜電描記術%小鼠
시망막변성%N-아초기-N-갑기뇨%현미경검사,전자%시망막전묘기술%소서
Retinal degeneration%N-Nitroso-N-methylurea%Microscopy,electron%Electroretinogram%Mice
目的 探讨N-亚硝基-N-甲基脲(MNU)诱导的小鼠视网膜退行性病变的形态和电生理特征.方法 实验研究.32只5周龄的成年C57/BL小鼠随机分为对照组和MNU造模组,分别腹腔注射0.9％氯化钠溶液和MNU (60 mg·kg-1).在给药后3、7和14 d取视网膜,免疫荧光染色观察光感受器形态、氧化损伤情况和神经节细胞的数量.电镜观察细胞核、线粒体和带状突触(synaptic ribbon)的超微结构.Ganzfeld视网膜电图(ERG)检测暗适应最大混合反应.采用独立样本t检验进行数据分析.结果 免疫荧光显示:注射MNU后外核层(ONL)的厚度逐渐减小,OPN lSW标记的光感受器细胞的外节(0S)逐渐损伤,GFAP标记的Müller细胞增生明显,ONL层硝基酪氨酸(Nitrotyrosine)着色增多提示MNU能导致氧化损伤.Bm-3a标记的神经节细胞数量减少不明显.扫描电子显微镜显示:MNU处理后,光感受器细胞的核呈浓染色,线粒体和带状突触消失.ERG结果显示:MNU处理后3d最大混合反应的a波和b波振幅下降.结论 MNU导致视网膜外核层和外丛状层的凋亡,电生理表现和视网膜色素变性疾病一致.
目的 探討N-亞硝基-N-甲基脲(MNU)誘導的小鼠視網膜退行性病變的形態和電生理特徵.方法 實驗研究.32隻5週齡的成年C57/BL小鼠隨機分為對照組和MNU造模組,分彆腹腔註射0.9％氯化鈉溶液和MNU (60 mg·kg-1).在給藥後3、7和14 d取視網膜,免疫熒光染色觀察光感受器形態、氧化損傷情況和神經節細胞的數量.電鏡觀察細胞覈、線粒體和帶狀突觸(synaptic ribbon)的超微結構.Ganzfeld視網膜電圖(ERG)檢測暗適應最大混閤反應.採用獨立樣本t檢驗進行數據分析.結果 免疫熒光顯示:註射MNU後外覈層(ONL)的厚度逐漸減小,OPN lSW標記的光感受器細胞的外節(0S)逐漸損傷,GFAP標記的Müller細胞增生明顯,ONL層硝基酪氨痠(Nitrotyrosine)著色增多提示MNU能導緻氧化損傷.Bm-3a標記的神經節細胞數量減少不明顯.掃描電子顯微鏡顯示:MNU處理後,光感受器細胞的覈呈濃染色,線粒體和帶狀突觸消失.ERG結果顯示:MNU處理後3d最大混閤反應的a波和b波振幅下降.結論 MNU導緻視網膜外覈層和外叢狀層的凋亡,電生理錶現和視網膜色素變性疾病一緻.
목적 탐토N-아초기-N-갑기뇨(MNU)유도적소서시망막퇴행성병변적형태화전생리특정.방법 실험연구.32지5주령적성년C57/BL소서수궤분위대조조화MNU조모조,분별복강주사0.9％록화납용액화MNU (60 mg·kg-1).재급약후3、7화14 d취시망막,면역형광염색관찰광감수기형태、양화손상정황화신경절세포적수량.전경관찰세포핵、선립체화대상돌촉(synaptic ribbon)적초미결구.Ganzfeld시망막전도(ERG)검측암괄응최대혼합반응.채용독립양본t검험진행수거분석.결과 면역형광현시:주사MNU후외핵층(ONL)적후도축점감소,OPN lSW표기적광감수기세포적외절(0S)축점손상,GFAP표기적Müller세포증생명현,ONL층초기락안산(Nitrotyrosine)착색증다제시MNU능도치양화손상.Bm-3a표기적신경절세포수량감소불명현.소묘전자현미경현시:MNU처리후,광감수기세포적핵정농염색,선립체화대상돌촉소실.ERG결과현시:MNU처리후3d최대혼합반응적a파화b파진폭하강.결론 MNU도치시망막외핵층화외총상층적조망,전생리표현화시망막색소변성질병일치.
Objective To explore the characteristics of N-Nitroso-N-methylurea (MNU) induced retinal degeneration in mice.Methods Experimental study.Thirty-two C57/BL mice (5 weeks) in total were randomly divided into two groups:a control group and a MNU group.Sixty mg·kg-1 of MNU was intraperitonealy injected in mice,leading to damage of retinal photoreceptors,while the control group was given physiological saline.The eyes of each group were enucleated at 3 d,7 d and 14 d after injection.The slides were stained by immunofluorescence labeling to observe the morphological changes to the photoreceptors in MNU-induced retinal damage.Retinal fiat-mount immunofluorescence was performed to observe the change in ganglion cells.The ultrastructures of the nucleus,mitochondria and synaptic ribbon were observed under transmission electron microscope.Ganzfeld electroretinogram ERG was used to test the electrophysiological changes.The data were analyzed using independent t test.Results After MNU administration,immunofluorescence labeling showed a gradual decrease in the thickness of the outer nuclear layer (ONL),the loss of photoreceptors labeled with anti-OPN1SW,reactive gliosis with anti-glial fibrillary acidic protein (GFAP) and oxidative damage in photoreceptors labeled with anti-nitrosylation.Brn-3a labeled ganglion cells did not appear to be lost two weeks after MNU administration.Electron microscopy showed concentrated coloring of the nucleus in the photoreceptor nucleus and the disappearance of mitochondria and metabolic synaptic ribbons after MNU administration.ERGs revealed that mice with MNU treatment showed a-waves and b-waves with reduced amplitudes.Conclusion MNU induced retinal apoptosis of ONL and outer piexiform layer.MNU administrated mice show similar electrophysiological results to retintis pigmentosa.
