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Title: [A preliminary study on the safety of berberine solution in rabbit eyes with topical application].
Author: Zhang Y, Bai F, Tao H.
Journal: Zhonghua Yan Ke Za Zhi; 2020 Feb 11; 56(2):131-137. PubMed ID: 32074824.
Abstract:
Objective: To study the safety of topical berberine solution in rabbit eyes and its effect on corneal epithelial repair in rabbit eyes. Methods: Experimental Study. Ninety-two Japanese rabbits were randomly divided into two groups by random number table method: the general group (32 rabbits, 64 eyes) and the corneal injury group (60 rabbits, 60 eyes).The general groups were further divided into 4 groups by random number table method, and each group has 8 rabbits (16 eyes). According to the administration of deionized water or 0.5, 1.0, 1.5 mg/ml berberine solution, they were divided into the general control group and the general A, B, and C group. Dosing with both eyes, each eye was given a single dose, and then it was given multiple times for 4 weeks after observation for 72h. After the corneal epithelium injury model made in the right eye of rabbits in the corneal injury groups, they were divided into a corneal injury control group and a corneal injury group A, B, and C according to the administration of deionized water or 0.5, 1.0, 1.5 mg/ml berberine solution. there were 5 rabbits (15 eyes) in each group, and the solutions were given continuously for 1 week. The rabbits in the general group were observed their behavioral changes, ocular surface and iris were scored by Draize eye irritation test scoring system. IOP was measured at different time points. Electroretinogram (ERG) was used to detect b-wave amplitude. In the corneal injury group, corneal epithelial defect repairment was observed at 1, 2, 3, 4, 5, 6, and 7 days after the corneal defect. Corneal histopathology observation after discontinuation of all rabbits. The pH value of rabbit tears was described by the paired t test, and the score of Draize eye irritation test were described by the rank-sum test. The analysis of variance and SNK-q were used for IOP, electroretinogram b-wave amplitude, corneal epithelial injury area and repair time. Results: No abnormal behavior was observed in the general group rabbits after single and multiple administration. There was no significant difference in the Draize eye irritation score among the general control group and the general group A, B, C at 1, 2, and 4 weeks of multiple administrations. Among them, the Draize eye irritation score of the general group C was 7 (0, 12), 6 (0, 10), 6 (0, 16) points (χ(2)=1.640, 0.265, 1.963, 1.381; P>0.05).There were no significant difference in IOP at different times among the general control group and the general group A, B, C at different times (F=0.065, 0.292, 0.015, 0.041; P>0.05). Before multiple administrations and after administration at 2, 4 weeks, the b-wave amplitudes of the general control group were (127.75±17.12), (129.18±15.83), (128.81±13.58) μV, and the general group A were (130.68).±18.75), (131.38±16.96), (130.62±12.18) μV,and the general group B were (128.00±16.74), (128.44±16.64), (129.06±16.16) μV, and the general group C were (131.81±19.37), (132.13±18.36), (129.94±12.60) μV. There was no statistically significant difference in b-wave amplitude in the groups at different times before and after administration (F=0.037, 0.011, 0.017, 0.702; P>0.05). There was no significant difference in the results of corneal histopathology among the general control group and the general group A, B, C. The area of corneal epithelial defect in each corneal injury group was statistically significant at different time (F=5.316, 25.864, 127.613; P<0.05). The corneal injury control group compared with the corneal injury group A, B, C, the corneal epithelial defect area in the corneal injury group C was significantly larger than the other three groups, with statistical differences (q=5.153, 10.313, 6.976; P<0.05). The repair time of corneal epithelial in control group and the group A,B,C of corneal injury were (83.0±1.85), (82.9±2.07), (83.7±2.09) and (101.6±2.20) h. The corneal epithelium defect repair time in group C was longer and the difference was statistically significant (F=301.437, P=0.000). Comparing the corneal injury control group and corneal injury group A and B, there was no statistical difference in the repair time of corneal epithelial defect (F=0.813, P=0.450). After repair, there was no significant difference in the pathological results of the corneal tissue between the corneal injury groups. Conclusions: Berberine solution in rabbit eyes with topical application was safety, and has no obvious toxic effect on the ocular surface and ERG of normal rabbits. 1.5 mg/ml berberine solution delayed the repair of experimental corneal epithelial defect, but had no effect on the integrity of corneal tissue after repair. (Chin J Ophthalmol, 2020, 56: 131-137). 目的： 研究兔眼部局部应用小檗碱溶液的安全性及其对兔眼角膜上皮修复的影响。 方法： 实验研究。日本大耳白兔92只，随机数字表法分为一般情况组（32只兔，64只眼）和角膜损伤组（60只兔，60只眼）。一般情况组再用随机数字表法分为4个组，每组8只兔（16只眼），根据给予去离子水或0.5、1.0、1.5 mg/ml小檗碱溶液，分为一般情况对照组及一般情况A、B、C组；双眼给药，每只眼先单次给药，观察72 h后再连续4周多次给药。角膜损伤组兔右眼制作角膜上皮损伤模型后，根据给予去离子水或0.5、1.0、1.5 mg/ml小檗碱溶液，分为角膜损伤对照组及角膜损伤A、B、C组，每组15只兔（15只眼），连续给药1周。一般情况组兔进行行为学观察，并应用Draize眼刺激性评分系统，对兔角膜、结膜、虹膜受累的面积、程度和反应进行评分，测量不同时间眼压，视网膜电图（ERG）检测b波振幅。角膜损伤组兔于角膜缺损后1、2、3、4、5、6、7 d观察角膜上皮缺损修复情况。所有兔停药后进行角膜组织病理学观察。兔泪液pH值比较采用配对t检验，Draize眼刺激性评分采用秩和检验，眼压、视网膜电图b波振幅及角膜上皮损伤面积和修复时间的多组比较采用方差分析及SNK-q检验。 结果： 单次给药和多次给药后一般情况组兔均无明显异常行为。一般情况对照及A、B、C组在多次给药1、2、4周时Draize眼刺激评分差异均无统计学意义，其中一般情况C组的Draize眼刺激评分分别为7（0，12）、6（0，10）、6（0，16）分（χ(2)=1.640，0.265，1.963，1.381；P>0.05）。一般情况对照及A、B、C组在多次给药前后不同时间的眼压差异均无统计学意义（F=0.065，0.292，0.015，0.041；P>0.05）。在多次给药前及给药后2、4周一般情况对照组的b波振幅分别为（127.75±17.12）、（129.18±15.83）、（128.81±13.58）μV，一般情况A组为（130.68±18.75）、（131.38±16.96）、（130.62±12.18）μV，一般情况B组为（128.00±16.74）、（128.44±16.64）、（129.06±16.16）μV，一般情况C组为（131.81±19.37）、（132.13±18.36）、（129.94±12.60）μV，各组在给药前后不同时间b波振幅差异均无统计学意义（F=0.037，0.011，0.017，0.702；P>0.05）。一般情况对照组与一般情况A、B、C组角膜组织病理学检查结果无明显差异。角膜损伤各组在不同时间的角膜上皮缺损面积有统计学意义（F=5.316，25.864，127.613；P<0.05），角膜损伤对照组与角膜损伤A、B、C组之间两两比较，角膜损伤C组的角膜上皮缺损面积明显大于其他3个组，差异具有统计学意义（q=5.153，10.313，6.976；P<0.05）。角膜损伤对照及A、B、C组的修复愈合时间为（83.0±1.85）、（82.9±2.07）、（83.7±2.09）和（101.6±2.20）h，角膜损伤C组角膜上皮缺损修复时间长，差异有统计学意义（F=301.437，P=0.000），角膜损伤对照组及角膜损伤A、B组之间比较，角膜上皮缺损修复时间差异无统计学意义（F=0.813，P=0.450）；修复结束后，角膜损伤组之间角膜组织的病理学检查结果无明显差异。 结论： 兔眼局部应用小檗碱溶液较安全，兔眼眼表未见明显的毒性反应，对视网膜未见明显影响。1.5 mg/ml浓度小檗碱溶液可延迟兔眼角膜上皮缺损修复，但对修复后的角膜组织结构的完整性无影响。（中华眼科杂志，2020，56：131-137）.

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