兔眼角膜着染对眼刺激性试验结果评价的影响

doi:10.12300/j.issn.1674-5817.2022.169

实验动物与比较医学 ›› 2023, Vol. 43 ›› Issue (3): 307-313.DOI: 10.12300/j.issn.1674-5817.2022.169

兔眼角膜着染对眼刺激性试验结果评价的影响

徐红花¹^,²(), 金天¹^,², 王海¹^,², 沈梦莹¹^,², 王瑞¹^,², 周艺佳¹^,², 谭莹¹^,²()()

^1.云南省药物研究所, 昆明 650111
^2.云南省中药和民族药新药创制企业重点实验室, 昆明 650111

收稿日期:2022-11-02 修回日期:2023-02-08 出版日期:2023-06-25 发布日期:2023-07-18
通讯作者: 谭莹（1979—），女，博士，高级工程师，研究方向：药物安全性评价研究。E-mail： tanying@ynby.cn。ORCID： 0009-0007-3084-9020
作者简介:徐红花（1978—），女，本科，高级工程师，主要从事药物安全性评价研究。E-mail： 601139654@qq.com
基金资助:
云南省药物研究所第三方公共技术服务平台能力提升建设(202104AR040002)

Influence of Corneal Staining in Rabbits on the Evaluation of Eye Irritation Test Results

Honghua XU¹^,²(), Tian JIN¹^,², Hai WANG¹^,², Mengying SHEN¹^,², Rui WANG¹^,², Yijia ZHOU¹^,², Ying TAN¹^,²()()

^1.Yunnan Institute of Materia Medica, Kunming 650111, China
^2.Yunnan Province Company Key Laboratory for TCM and Ethnic Drug of New Drug Creation, Kunming 650111, China

Received:2022-11-02 Revised:2023-02-08 Published:2023-06-25 Online:2023-07-18
Contact: TAN Ying (ORCID: 0009-0007-3084-9020), E-mail: tanying@ynby.cn

摘要/Abstract

摘要：

目的观察正常实验兔眼经荧光素钠染色后的着染现象对眼刺激性试验结果评价的影响。方法在进行针对氯化钠滴眼液、四味珍层冰硼滴眼液、玻璃酸钠滴眼液、色甘酸钠滴眼液和复方门冬维甘滴眼液的实验兔眼刺激性试验（每组4只，雌雄各半）中，将给予生理盐水的兔左眼（自身阴性对照）和给予药物制剂的右眼（给药实验）进行1%荧光素钠染色，然后用裂隙灯显微镜连续31 d观察兔眼刺激反应并评分，记录兔角膜上皮着染的形态变化并计算着染发生率。观察结束后对眼球及哈氏腺进行组织病理学检查，并将左眼着染发生率检查结果与给予相应药物的右眼进行统计学对比分析。结果给予生理盐水的左眼和给予药物制剂的右眼均无眼刺激反应，评分均为0，角膜着染总发生率分别为3%、1%，左右眼每天和总着染发生率无明显差异（P>0.05）。角膜上皮着染形态有单点状着染、散在点状着染、局限性着染和大片融合着染的无规律变化，眼球及哈氏腺未见组织病理学改变，结果评价为无刺激性。结论实验兔眼无规律间断出现的角膜上皮着染对眼刺激性试验的结果评价无影响。

关键词: 实验兔, 角膜, 着染, 眼刺激性试验, 结果评价

Abstract:

Objective To observe the influence of the staining phenomenon after fluorescein sodium staining on eye irritation in normal rabbits. Methods In the experimental rabbit eye irritation test conducted with sodium chloride eye drops, Siwei Zhenceng Bingpeng eye drops, sodium hyaluronate eye drops, sodium cromoglycate eye drops, and compound aspartate eye drops (4 in each group, half male and half female), the left eyes of rabbits were administered normal saline (self-negative control) and the right eyes were administered the experimental medicine; the eyes were stained with 1% sodium fluorescein, and eye irritation was observed and scored using slit lamp microscope for 31 days. Morphological changes of corneal epithelial staining were recorded and the incidence of staining was calculated. After the observation, the eyeballs and Hasselblad glands were examined histopathologically, and the staining rate of the left eye was compared with that of the right eye which was administered the corresponding medicine. Results Neither eye had any irritation symptoms; the scores were 0, and the total incidences of corneal staining were 3% (left) and 1% (right), respectively. There was no significant difference between the two groups (P > 0.05). Corneal epithelial staining showed single-spot staining, scattered dot, localized, or large areas of fusion staining. No histopathological changes were found in the eyeballs or Hasselblad glands, and the results were evaluated as non-irritative. Conclusion The irregularity of corneal epithelial staining in rabbits did not influence the results of the ocular irritation test.

Key words: Rabbits, Cornea, Staining, Eye irritation test, Result evaluation

中图分类号:

R-332

徐红花, 金天, 王海, 沈梦莹, 王瑞, 周艺佳, 谭莹. 兔眼角膜着染对眼刺激性试验结果评价的影响[J]. 实验动物与比较医学, 2023, 43(3): 307-313.

Honghua XU, Tian JIN, Hai WANG, Mengying SHEN, Rui WANG, Yijia ZHOU, Ying TAN. Influence of Corneal Staining in Rabbits on the Evaluation of Eye Irritation Test Results[J]. Laboratory Animal and Comparative Medicine, 2023, 43(3): 307-313.

图/表 4

表1 兔眼刺激反应分值标准

Table 1 The standard scores for eye ocular irritation in rabbits

刺激部位 Site of irritation	眼刺激反应 Reaction to eye irritation	分值 Score
角膜Cornea	无混浊	0
	散在或弥漫性混浊，虹膜清晰可见	1
	半透明区易分辨，虹膜模糊不清	2
	出现灰白色半透明区，虹膜细节不清，瞳孔大小勉强可见	3
	角膜不透明，虹膜无法辨认	4
虹膜Iris	正常	0
	皱褶明显加深、充血、肿胀，角膜周围轻度充血，瞳孔对光仍有反应	1
	出血，肉眼可见坏死，对光无反应（或其中一种）	2
结膜Conjunctiva	血管正常	0
	血管充血呈鲜红色	1
	血管充血呈深红色，血管不易分辨	2
	弥漫性充血呈紫红色	3
	无水肿	0
	轻微水肿（含眼睑）	1
	明显水肿伴部分眼睑外翻	2
	水肿至眼睑近半闭合	3
	水肿至眼睑超过半闭合	4
	无分泌物	0
	少量分泌物	1
	分泌物使眼睑和睫毛潮湿或黏着	2
	分泌物使整个眼区潮湿或黏着	3
最大总积分Maximum total integral value		16

表2 观察期间各组实验兔左右眼角膜上皮着染出现的时间和发生率

Table 2 The time and incidence of corneal epithelial staining in the left and right eyes of the rabbits in each group during the observation period

组别 Test group	观察部位 Observed position	每天发生率/% Daily incidence/%											总发生率/% Total incidence/%
组别 Test group	观察部位 Observed position	D1	D3	D9	D10	D12	D17	D18	D20	D24	D28	D31	总发生率/% Total incidence/%
氯化钠滴眼液组（n=4） Sodium chloride eye drops	左眼	0	0	25	0	25	0	0	25	25	0	0	3.2
氯化钠滴眼液组（n=4） Sodium chloride eye drops	右眼	0	25	0	0	0	25	0	0	0	25	0	2.4
四味珍层冰硼滴眼液组（n=4） Siwei Zhenceng Bingpeng eye drops	左眼	0	0	0	25	25	0	0	25	0	0	0	2.4
四味珍层冰硼滴眼液组（n=4） Siwei Zhenceng Bingpeng eye drops	右眼	0	25	0	0	0	0	25	0	0	0	0	1.6
玻璃酸钠滴眼液组（n=4） Sodium hyaluronate eye drops	左眼	0	0	25	0	25	0	0	25	0	0	0	2.4
玻璃酸钠滴眼液组（n=4） Sodium hyaluronate eye drops	右眼	0	0	0	0	0	25	0	0	0	0	0	0.8
色甘酸钠滴眼液组（n=4） Sodium cromoglycate eye drops	左眼	0	0	0	0	25	0	0	0	25	0	25	2.4
色甘酸钠滴眼液组（n=4） Sodium cromoglycate eye drops	右眼	0	0	0	0	0	0	25	0	0	0	0	0.8
复方门冬维甘滴眼液组（n=4） Compound aspartate eye drops	左眼	0	0	0	25	0	0	0	25	0	0	25	2.4
复方门冬维甘滴眼液组（n=4） Compound aspartate eye drops	右眼	0	0	0	0	0	0	0	0	0	25	0	0.8
全部实验兔（n=20） All experimental rabbits	左眼	0	0	10	10	20	0	0	20	10	0	10	3.0
全部实验兔（n=20） All experimental rabbits	右眼	0	10	0	0	0	10	10	0	0	10	0	1.0

图1 观察期间实验兔的左右眼角膜上皮着染出现的时间和形态汇总注：A，不同时间的右眼（给药）着染形态汇总：第3天（D3），散在点状着染；第17、18、28天（D17、D18、D28），局限性着染。B，不同时间的左眼（阴性对照）着染形态汇总：第9天（D9），单点状着染；第10、12、20、30天（D10、D12、D20、D30），局限性着染；第24天（D24），大片融合着染。

Figure 1 Summary of the timing and morphology of corneal epithelial staining in the left and right eyes of rabbits during the observation period.Note：(A) The morphology of staining in the right eye at different times after administration is summarized: D3, scattered dot; D17, D18, and D28, localized dot. (B) The morphology of staining in the left eye at different times after administration is summarized: D9, single dot; D10, D12, D20, and D30, localized dot; D24, large fusion.

图2 实验兔染色检查31 d后眼球及哈氏腺组织病理学观察（HE染色，×20）注：A，左眼角膜正常；B，右眼角膜正常；C，左眼视网膜正常；D，右眼视网膜正常；E，左眼哈氏腺正常；F，右眼哈氏腺正常。各图中比例尺大小为100 μm。左右眼每个组织标本样本数各为20。

Figure 2 The histopathology of the eyeballs and Harderian glands were observed after 30 days of staining （HE staining， ×20）Note： (A) Normal cornea in the left eye; (B) Normal cornea in the right eye; (C) Normal retina in the left eye; (D) Normal retina in the right eye; (E) Normal Hasselblad gland in the left eye; (F) Normal Hasselblad gland in the right eye. The scale size of each figure is 100 μm. A total of 20 tissue samples were collected from each eye.

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兔眼角膜着染对眼刺激性试验结果评价的影响

Influence of Corneal Staining in Rabbits on the Evaluation of Eye Irritation Test Results

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