【摘要】 目的：选择性干扰黄斑区Müller细胞，试图建立特发性黄斑旁毛细血管扩张症(idiopathic perifoveal telangiectasis,IPT)动物模型。方法：恒河猴12只随机分为6组，前3组单眼视网膜下注入DLαAAA(DLαaminoadipic acid)5，10，50mmol/L 30μL，后3组单眼玻璃体腔分别注入DLαAAA 16，50，80mmol/L 100μL。术前1wk及术后6，12wk行眼底彩色照相、荧光素眼底血管造影、黄斑自发荧光、光学相干断层成像(optical coherence tomography，OCT)、多焦视网膜电图(multifocal roretinogram，mfERG)检测，并摘除眼球行光镜检查。结果：视网膜下5，10mmol/L DLαAAA及玻璃体腔50mmol/L DLαAAA，给药后6wk均出现IPT且OCT和光镜亦有相应病理改变;视网膜下50mmol/L DLαAAA，及玻璃体腔80mmol/L DLαAAA，病理损伤严重但未出现IPT。结论：视网膜下5～10mmol/L DLαAAA、玻璃体腔50mmol/L DLαAAA均可诱发IPT。
【关键词】 恒河猴;Müller细胞;DLαAAA;IPT;视网膜下给药
Induction of idiopathic perifoveal telangiectasis by DLαAAA in rhesus monkeysJun Zhang, YunTao Hu, ZhiZhong Ma, XunLun ShengEye Center of Peking University, Beijing 100083, China; Department of Ophthalmology, Affiliated Hospital of Ningxia Medical University, Yinchuan 750004, Ningxia Hui Autonomous Region, China;Department of Ophthalmology, the First Hospital of Qingdao Development Zone, Qingdao 050051, Shandong Province, ChinaAbstractAIM: Selective disturbance of macular Müller cells on the neural retina and retinal vasculature in nonhuman primates. Try to establish the animal model of idiopathic perifoveal telangiectasis(IPT). METHODS: Twelve rhesus monkeys (24 eyes) were divided into 6 groups randomly. In the former 3 groups, DLαaminoadipic acid (DLαAAA) solution 30μL of 5, 10, 50mmol/L were injected subretinally respectively, one eye for each rhesus monkey, as 3 experimental groups. The control groups were injected subretinally PBS 30μL in other eyes. In the later 3 groups, DLαAAA solution 100μL of 16, 50, 80mmol/L were injected intravitreally respectively, one eye for each rhesus monkey, as 3 experimental groups. The control groups were injected intravitreally PBS 100μL in other eyes. All eyes were examined by fundus color photography, fluorescein angiography, macular autofluorescence, optical coherence tomography(OCT), multifocal electroretinography(mfERG) and optical microscopy at 1st week before the operation and 6th, 12nd week after the operation.
RESULTS: After 6th week of operation, IPT were happened in the groups of subretinal 5mmol/L, 10mmol/L and the group of intravitreal 50mmol/L. In the same concentration groups, corresponding pathological changes were found by OCT and optical microscopy. The group of subretinal 50mmol/L and the group of intravitreal 80mmol/L response a more serious pathological changes, but IPT has not occurred in those groups. CONCLUSION: DLαAAA as ah agentspecific interference of Müller cells in retina, the concentration between subretinal 5mmol/L to 10mmol/L and the concentration intravitreal 50mmol/L could induce IPT.
KEYWORDS: rhesus monkeys; Müller cells; DLαAAA; IPT; subretinal injection0 引言
特发性黄斑旁毛细血管扩张症(idiopathic perifoveal telangiectasis，IPT)是一种罕见的潜在致盲性眼病[1]。由于病源稀少又缺乏病因学的研究。IPT动物模型的创建，无疑成为攻克IPT的研究热点。通过近期研究，发现IPT最初的异常发生于视网膜的神经和胶质成分——Müller细胞[2]。我们设想通过选择性干扰黄斑区Müller细胞以建立IPT动物模型，旨在为今后IPT的实验研究及临床研究提供依据。
1 材料和方法
1.1 材料
成年恒河猴12只，体质量6±1.5kg，雌雄不限，购自北京大学医学部动物实验中心。
1.2 方法
每只动物随机取单眼分成6组，每组2眼。前3组视网膜下分别注入5，10，50mmol/L DLαAAA (sigmaaldrich)30μL，其余对侧眼视网膜下均注入PBS 30μL作为对照。后3组玻璃体腔内分别注入16，50，80mmol/L DLαAAA 100μL，其余对侧眼玻璃体腔内均注入PBS 100μL作为对照。溶液以PBS配制。速眠新、盐酸氯胺酮1∶1配比全身麻醉，贝诺喜滴眼液表面麻醉。前3组复方托吡咔胺点眼以散大瞳孔，于角膜缘后2mm处，颞上方进导光纤维照明，鼻上方进自制视网膜下注药系统，将DLαAAA 30μL注入黄斑颞下方2CD处，对照组注入PBS 30μL;后3组于颞上方角膜缘后2mm处30G针头穿刺向玻璃体腔内注入DLαAAA 100μL，对照组注入PBS 100μL。每组注液后均前房穿刺放出少许房水降低眼压至正常。每眼注药前1wk和注药后6，12wk行眼底彩色照相、荧光素眼底血管造影、黄斑自发荧光、光学相干断层成像(optical coherence tomography，OCT)、多焦视网膜电图(multifocal electroretinogram，mfERG)检测;术后12wk处死动物摘除眼球，40g/L多聚甲醛溶液固定，视网膜组织石蜡包埋，切片5μm HE染色行光镜检查。
统计学分析：用SPSS 13.0统计学软件对数据(均值±标准差)进行处理。
2 结果
所有实验猴眼均未发生眼内炎及玻璃体混浊。眼底彩色照相所有眼均无玻璃体积血，但前3组有明显视网膜下浅脱离。视网膜下5，10mmol/L给药组光镜下可见外丛状层破坏明显;50mmol/L组可见感光细胞层和外丛状层破坏明显(图1);玻璃体腔16mmol/L给药组未见明显异常;50，80mmol/L组感光细胞层和外丛状层破坏明显。
2.1 FFA检查
各组均可见黄斑旁微血管瘤，其中视网膜下5，10mmol/L给药组及玻璃体腔50mmol/L给药组出现IPT表现(图2)。
2.2 OCT检查
术后12wk视网膜下药液被完全吸收。其中视网膜下5mmol/L给药组可见小血管扩张;10，50mmol/L组可见轻微黄斑囊样扩张;玻璃体腔16mmol/L给药组未见明显异常;50mmol/L组可见黄斑前膜(图3);80mmol/L组可见小血管扩张。
2.3 mfERG检查
各实验组黄斑区b波振幅的变化随术后时间的进展呈下降趋势(图4)。
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