摘要
目的探讨兔皮下注射氢溴酸东莨菪碱建立干眼模型的有效性和眼表病理损害机制。设计实验研究研究对象健康新西兰大白兔9只。方法将9只白兔随机分为三组,每组3只。实验组皮下注射氢溴酸东莨菪碱,其中低剂量组(L,1.0 mg/次)、高剂量组(H,2.0 mg/次),对照组注射等量生理盐水。注射时间点为每天8点、11点、14点和18点,连续30天。主要指标不同时间点用药前后泪液分泌量、泪膜破裂时间,角膜荧光素染色情况,结膜印迹联合PAS染色评价眼表上皮细胞表型。观察期末取角结膜、泪腺组织行HE染色光镜下观察。结果酚红棉线法测得高剂量(H)组注射前泪液分泌量为(16.25±2.299)mm,第21d时下降为(6.75±1.982)mm,对照组为(16.50±2.619)mm,差异具有统计学意义(P=0.000<0.01),同期低剂量组(L)为(15.00±2.390)mm,与对照组相比无明显差异。H组第3d出现BUT缩短,L组则第7d发生BUT的变化,此后BUT值均低于5s,对照组BUT大于20s。实验组均出现角膜荧光素染色阳性,H组出现时间早(3 d),持续时间长。L组和H组PAS染色与对照组相比,杯状细胞形态紊乱、核淡染,部分核固缩,L组和H组之间无明显差异。HE染色显示H组泪腺周围见散在淋巴细胞和腺腔凋亡性改变,角膜上皮呈重度瘤样增生基质层变性伴水肿,结膜杯状细胞反应性增多,固有层见淋巴细胞浸润数量增加。L组角膜浅层上皮轻度增生,细胞排列稍紊乱,结膜和泪腺组织病理改变轻微。结论兔皮下注射氢溴酸东莨菪碱,4次/d,2.0 mg/次,可有效维持药物外周浓度,抑制泪腺分泌,引起角结膜上皮损伤,成功建立以泪液缺乏为主,炎症反应参与的干眼动物模型,为进行干眼发病机制研究和药物干预提供了理想的实验平台。
Objective To evaluate the characteristic of rabbit tear-deficient dry eye model by subcutaneous injection of scopolamine hydrobromide and explore the mechanism of ocular surface. Design Experimental study. Participants Nine healthy New Zealand white rabbits. Methods The rabbits were randomly divided into three groups, 3 rabbits in each group. Subcutaneous injection of scopolamine hydrobromide was made in experimental group including 1.0mg in low dose group, 2.0mg in high dose group and normal saline in control group, at 8am, llam, 14pro and 18pm each day for 30 days. Main Outcome Measures Schirmer I test, tear membrane break-up time（BUT）, corneal fiuorescein staining and conjunctival impression cytology test with PAS staining to evaluate the phe- notype of epithelia ceils were performed before and 7, 14, 21, 30 days ＇after injection. On the thirtieth day, cornea, conjunctiva and lacrimal glands tissues biopsy were carried out by H-E staining . Results Schirmer I test by phenol red thread test of H group was （16.25＋2.299）mm before injection and decreased to （6.75 ±1.982）mm on the 21st day, with a statistically difference （P=0.000〈0.01） compared to （ 16.50±2.619 ）mm of the control group, however, it was （ 15.00±2.390）mm of L group with no statistically difference. BUT decreased at the 3rd day in high dose group and the 7th Day in low dose group after which it kept shorter than 5s, BUT kept greater than 20s in control group. Flurescein staining of corneal were found in two experimental groups. It was initially observed in H group at the 3rd day and last for a long time. Compared with the control group, the morphology of goblet cells were irregular and nucleus were slightly stained, some of which were pyknotic in two experimental groups. There was no significant difference between H and L group. H-E stainning showed that lymphocyte dispersedly distributed around lacrimal gland, apoptosis in glandular cavity, severe tumor-like hyperplasia in corneal epithelium, degeneration with edema in corneal stroma layer, conjunctival goblet cell increased reactively and increased the number of cell infiltration in lamina propria in H group, Slight hyperptasia, cell disarrangement in corneal and mild histopathology changes in conjunctiva and lacrimal gland. Conclusions Subcutaneous injection of scopolamine hydrobromide, 2mg every time for 4 times a day can effectively resulting in lacrimal secretion decrease and damage to epithelium of coneal and conjunctiva, which mainly caused by tear deficient and inflammation may also participate in.
出处
《眼科》
CAS
2009年第6期404-409,共6页
Ophthalmology in China
基金
广东省自然科学基金(7001679)
教育部新教师基金(20070558282)
