牛磺酸对苯扎氯铵损伤大鼠眼角膜内皮细胞无保护作用

doi:10.12300/j.issn.1674-5817.2022.034

摘要/Abstract

摘要：

目的观察2%牛磺酸对苯扎氯铵损伤大鼠眼角膜内皮细胞的保护作用。方法取3只SPF级SD大鼠（共6只眼），每只眼制备6块角膜内皮层和后弹力层组织片，随机分为3组。用组织块培养法培养大鼠眼角膜内皮细胞1 d后，对照组仅加入2%牛磺酸溶液，实验组在加入2%牛磺酸溶液后再注入质量分数分别为0.01%和0.03%的苯扎氯铵溶液。继续培养1、2、4、5、6、8 d后倒置显微镜下观察各组大鼠眼角膜内皮细胞的生长情况，并用Wright染色后光学显微镜下观察内皮细胞形态。结果用0.01%苯扎氯铵和2%牛磺酸处理1 d的实验组大鼠眼角膜组织块边缘出现多角形的内皮细胞，胞体透明；2 d后多角形细胞增多，但没有出现细胞间的融合生长现象；3 d后多角形细胞减少，内皮细胞未见核分裂象；4 d后可见内皮细胞核着色加深，多角形细胞少见；5 d后内皮细胞数量减少，出现细胞体固缩死亡现象。用0.03%苯扎氯铵和2%牛磺酸处理1 d的实验组未观察到内皮细胞生长，胞体稀少散在。仅用2%牛磺酸处理的对照组1 d后组织块边缘出现多角形内皮细胞和少数类内皮多边形细胞；3 d后组织块边缘多角形细胞增多，而且有逐渐融合生长的现象；5 d后内皮细胞个数增多，而且细胞外形多呈六角形；8 d后内皮细胞形成大片状，胞体呈六角形或类圆形，出现细胞核分裂象。左右眼角膜内皮细胞生长均匀，在0.01%和0.03%苯扎氯铵处理的实验组以及对照组内左右眼内皮细胞形态均无明显差别。结论 2%牛磺酸对被不同浓度的苯扎氯铵液损伤的大鼠眼角膜内皮细胞无保护作用。

关键词: 牛磺酸, 苯扎氯铵, 眼角膜内皮细胞, 组织培养, SD大鼠

Abstract:

Objective To observe the protective effect of 2% taurine on corneal endothelial cells injured by benzalkonium chloride in rats. Methods Six piece of corneal endodermis and elastic layer tissue slices were prepared from 6 eyes of 3 SPF SD rats and randomly divided into three groups. The corneal endothelial cells of rats were cultured by tissue block culture for 1 day, then the control group cells were added with 2% taurine solution, while the experimental group cells were added with 2% taurine solution and 0.01% or 0.03% benzalkonium chloride solution. After 1, 2, 4, 5, 6 and 8 days of continuous culture, the growth of corneal endothelial cells in each group was observed under an inverted microscope, and the morphology of endothelial cells was observed under an optical microscope after Wright staining. Results Treated with 0.01% benzalkonium chloride and 2% taurine for 1 day, polygonal endothelial cells appeared on the edge of corneal tissue mass, and the cells were transparent. After 2 days, the number of polygonal cells increased, and there was no fusion growth between cells. After 3 days, the number of polygonal cells decreased and no mitotic signs were observed in endothelial cells. After 4 days, the endothelial nuclei were deeply stained and polygonal cells were rare. After 5 days, the number of endothelial cells decreased, and cell body shrinkage and death occurred. In the experimental group treated with 0.03% benzalammonium chloride and 2% taurine for 1 day, no endothelial cell growth was observed and the cells were sparsely-scattered. In control group, polygonal endothelial cells and a few endothelium-like polygon cells appeared at the edge of tissue blocks after 1 day. After 3 days, the number of polygonal cells at the edge of tissue blocks increased, and there was a phenomenon of gradual fusion growth. After 5 days, the number of endothelial cells increased, and the cells were mostly hexagonal. After 8 days, the endothelial cells formed large sheets, the cell bodies were hexagonal or round, and the nuclei were divided. The growth of corneal endothelial cells in the left and right eyes was uniform, and there was no significant difference in the morphology of the left and right eye endothelial cells in the 0.01% and 0.03% benzalammonium chloride treatment groups and the control group. Conclusion 2% taurine had no protective effect on corneal endothelial cells injured by benzalammonium chloride.

Key words: Taurine, Benzalkonium chloride, Corneal endothelial cells, Tissue culture, SD rats

中图分类号:

R-332

葛健, 孙静芬, 吴永杰. 牛磺酸对苯扎氯铵损伤大鼠眼角膜内皮细胞无保护作用[J]. 实验动物与比较医学, 2023, 43(1): 39-43.

Jian GE, Jingfen SUN, Yongjie WU. Taurine Has no Protective Effect on Rat Corneal Endothelial Cells Injured by Benzalkonium Chloride[J]. Laboratory Animal and Comparative Medicine, 2023, 43(1): 39-43.

图/表 1

图1 大鼠眼角膜内皮细胞培养生长情况和细胞形态（Wright染色，×40）注：A～C，用2%牛磺酸+0.01%苯扎氯铵处理的实验组1大鼠眼角膜内皮细胞：A显示培养第2天，多角形细胞增多，但没有细胞间的融合现象；B显示培养第4天，可见多角形细胞少，细胞固缩，内皮细胞稀少；C显示培养第5天，内皮细胞数减少，出现细胞体固缩死亡现象。D，用2%牛磺酸+0.03%苯扎氯铵处理的实验组2大鼠眼角膜内皮细胞，培养第1天，内皮细胞无生长。E～G，用2%牛磺酸（未加入苯扎氯铵）处理的对照组大鼠眼角膜内皮细胞：E显示培养第2天，出现多量的多角形细胞，而且有细胞聚集生长现象；F显示培养第4天，可见多量的多角细胞，内皮细胞增多，胞体规则且多呈六角形，内皮细胞成片状；G显示培养第5天，可见细胞核染色质浓染，具核分裂象。

Figure 1 Growth and morphology of rat corneal endothelial cells （Wright staining， ×40）Note： A-C, corneal endothelial cells of experimental group 1 treated with 2% taurine + 0.01% benzalkonium chloride: A showed on the second day of culture, the number of polygonal cells increased, while there was no fusion growth between cells; B showed on the 4th day of culture, polygonal cells pycnosis, the number of polygonal cells and corneal endothelial cells were rare; C showed after on the 5th day of culture, the number of endothelial cells decreased, and cell body shrinkage and death occurred; D, corneal endothelial cells of experimental group 2 treated with 2% taurine + 0.03% benzalkonium chloride, after cultured 1 day, no endothelial cell growth was observed. E-G, corneal endothelial cells of control group treated with 2% taurine: E showed that on the second day of culture, a large number of polygonal cells appeared, and there was cell aggregation and growth; F showed that on the 4th day of culture, a large number of polygonal cells and endothelial cells were observed, the cell body was regular and mostly hexagonal, and the endothelial cells were lamellar; G showed that on the 5th day of culture, the nuclear chromatin was heavily stained and the mitotic phase was observed.

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