Yu Fen-fen, Liu Xing, Zhong Yi-min, Mao Zhen, Guo Xin-xing, Li Mei, Cao Dan, Chen Xiang-xi
State Key Laboratory of Ophthalmology, Zhongshan Ophthalmic Center, Sun Yat-sen University, Guangzhou 510060, China.
State Key Laboratory of Ophthalmology, Zhongshan Ophthalmic Center, Sun Yat-sen University, Guangzhou 510060, China. Email:
Zhonghua Yan Ke Za Zhi. 2013 Nov;49(11):973-80.
To investigate the protective effects of sodium hyaluronate on ocular surface toxicity induced by a prolonged use of benzalkonium chloride-preserved Brimonidine eye drops.
Experimental study. Thirty adult female New Zealand rabbits were divided into three groups with randomized numbers design. Ten rabbits were treated with 0.2% Brimonidine eye drops and PBS (PBS group), the other ten rabbits with 0.2% Brimonidine combined with sodium hyaluronate eye drops (SH group), and control group received no treatment for 60 days. Schirmer test, fluorescein (FL) and Rose Bengal (RB) staining, conjunctival impression cytology specimens collecting were performed on day 0, 31, and 61. Apoptosis of conjunctival epithelium was detected by in situ terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) assay on day 61. Conjunctival inflammation was observed by hematoxylin eosin staining. Histomorphological changes of cornea and conjunctiva were observed by light microscopy, and scanning and transmission electron microscopy at day 61. Fluorescein and Rose Bengal scores were analysed by Kruskal-Wallis test. Schirmer scores, goblet cell density and inflammatory cells infiltration were analysed by repeated measures analysis of variance.
There were significant differences in fluorescein and Rose bengal (H = 22.031, 15.303, P < 0.01) staining among the groups on day 61. Compared with the control group (FL: 0, 0-1, RB: 0, 0-1), fluorescein and Rose Bengal scores were significantly (P < 0.001) increased in PBS group (FL: 1.5, 1-2, RB: 1, 1-2), whereas was significantly (P < 0.001) decreased in SH group (FL:0, 0-1 RB:1, 0-1) when compared to PBS group. There were significant differences in aqueous tear production and goblet cell density (F = 7.980, 14.545, both P < 0.01) among the groups on day 61. Compared with the control group [(9.43 ± 0.57) mm, (87.73 ± 2.34/HP)], Schirmer scores and goblet cell density were significantly (P < 0.01) reduced in PBS-treated group [(6.61 ± 0.38) mm, (68.06 ± 3.61)/HP], but significantly (P < 0.05) increased in SH-treated group [(8.75 ± 0.57) mm, (82.31 ± 1.64)/HP] compared with PBS-treated group. The number of inflammatory cells was significant difference (F = 56.306, P < 0.001) among the groups on day 61. Compared with the control group [(39.89 ± 2.03)/HP], inflammatory cells infiltration was significantly (P < 0.01) increased in both PBS [(73.18 ± 2.17)/HP] and SH groups [(48.79 ± 2.64)/HP], however, SH-treated group was significantly lowered when compared with PBS-treated group. In addition, decrease in apoptosis, complete microvilli and cell organelles were found in the corneal and conjunctival epithelial cells in SH-treated group.
Our results demonstrate that topical application of SH reduces the ocular toxicity and protect the ocular surface in the long term anti-glaucomatous medical therapies and may be considered as a vehicles or neutralizing material for future ocular application.
探讨透明质酸钠对长期使用含苯扎氯铵的溴莫尼定滴眼液所致眼表毒性的保护作用。
实验研究。将30只成年雌性新西兰兔随机分为三组。10只兔用0.2%溴莫尼定滴眼液和磷酸盐缓冲液（PBS组）处理，另外10只兔用0.2%溴莫尼定联合透明质酸钠滴眼液处理（SH组），对照组60天不做处理。在第0、31和61天进行泪液分泌试验、荧光素（FL）和孟加拉玫瑰红（RB）染色、采集结膜印迹细胞学标本。在第61天通过原位末端脱氧核苷酸转移酶dUTP缺口末端标记（TUNEL）法检测结膜上皮细胞凋亡。通过苏木精-伊红染色观察结膜炎症。在第61天通过光学显微镜、扫描电子显微镜和透射电子显微镜观察角膜和结膜的组织形态学变化。采用Kruskal-Wallis检验分析荧光素和孟加拉玫瑰红评分。采用重复测量方差分析分析泪液分泌试验评分、杯状细胞密度和炎症细胞浸润情况。
第61天，各组间荧光素和孟加拉玫瑰红染色有显著差异（H = 22.031, 15.303, P < 0.01）。与对照组（FL: 0, 0 - 1, RB: 0, 0 - 1）相比，PBS组荧光素和孟加拉玫瑰红评分显著升高（P < 0.001）（FL: 1.5, 1 - 2, RB: 1, 1 - 2），而SH组与PBS组相比显著降低（P < 0.001）（FL: 0, 0 - 1, RB: 1, 0 - 1）。第61天，各组间泪液分泌量和杯状细胞密度有显著差异（F = 7.980, 14.545, 均P < 0.01）。与对照组[（9.43 ± 0.57）mm, （87.73 ± 2.34/HP）]相比，PBS处理组泪液分泌试验评分和杯状细胞密度显著降低（P < 0.01）[（6.61 ± 0.38）mm, （68.06 ± 3.61）/HP]，但SH处理组与PBS处理组相比显著升高（P < 0.05）[（8.75 ± 0.57）mm, （82.31 ± 1.64）/HP]。第61天，各组间炎症细胞数量有显著差异（F = 56.306, P < 0.001）。与对照组[（39.89 ± 2.03）/HP]相比，PBS组[（73.18 ± 2.17）/HP]和SH组[（48.79 ± 2.64）/HP]炎症细胞浸润均显著增加（P < 0.
