噻吗心安对人角膜内皮细胞的影响作用研究  

Studies on the effect of timolol maleate on human corneal endothelial cells

作　　者：樊廷俊[1] 随蓓蓓[1] 王清扬[1] 温茜[1] 孙倩[1] 于苗苗[1] 葛源[1] 

机构地区：[1]中国海洋大学海洋生命学院角膜组织工程重点实验室,中国山东省青岛市266003

出　　处：《国际眼科杂志》2013年第6期1069-1072,共4页International Eye Science

基　　金：国家高技术研究发展863计划(No.2006AA02A132)~~

摘　　要：目的:揭示抗青光眼药物噻吗心安(timolol maleate)对体外培养人角膜内皮(HCE)细胞的影响作用,为其眼科临床安全用药提供实验依据。方法:用不同浓度噻吗心安处理体外培养的HCE细胞,在倒置显微镜下观察细胞的生长、增殖和形态变化,利用吖啶橙/溴化乙锭荧光双染色法检测质膜的通透性,利用琼脂糖凝胶电泳法检测DNA的断片化,利用透射电镜观察细胞的超微结构。结果:噻吗心安在0.15625～5g/L的浓度范围内均能不同程度地引起HCE细胞出现不同程度的生长缓慢、数量减少、胞质皱缩、胞内空泡化、变圆脱落、质膜通透性增大、染色质凝缩、DNA断片化和出现凋亡小体等典型的细胞凋亡特征,并具有浓度和时间依赖性。临床使用浓度2.5～5g/L的噻吗心安对HCE细胞的凋亡诱导作用最大,处理28h的凋亡率高达83.23%～96.71%。结论:噻吗心安在0.15625～5g/L的浓度范围内能显著诱导HCE细胞凋亡,临床使用浓度时对HCE细胞的毒性作用极大,在眼科临床中应谨慎使用。AIM ： To examine the effect of timolol maleate, a topical anti-glaucoma drug, on in vitro cultured human corneal endothelial （HCE） cells and lay foundations for its secure medication clinically. METHODS ： After in vitro cultured HCE cells treated with timolol maleate at different concentrations, the situation of cellular growth, proliferation, and morphology was checked under an inverted light microscope. And plasma membrane permeability, DNA fragmentations, and ultra- structure of HCE cells were detected by acridine orange/ ethidium bromide （AO/EB） double fluorescent staining, DNA agarose gel electrophoresis, and transmission electron microscopy （TEM）, respectively. RESULTS： After treated with timolol maleate at concentrations from 0. 15625g/L to 5g/L, in vitro cultured HCE cells showed typical characteristics of apoptosis, including growth retarding, number decreasing, cytoplasmic shrinking, vacuolation, falling off from well bottom of culture plate, plasma membrane permeabilityincreasing, chromatin condensation, DNA fragmentation, appearance of apoptotic body, and so on. The apoptosis- inducing effect of timolol maleate was in dose- and time- dependent manners. The greatest apoptosis- inducing effect of timolol maleate on HCE cells was found at the clinic medication concentration of 2. 5- 5g/L, and their induced apoptotic rate of HCE ceils at 28 hours reached 83.23% -96.71%, respectively.CONCLUSION. Timolol maleate within the concentration of0. 15625-5g/L has an obvious apoptosis-inducing effect on HCE cells, and has tremendous toxicity to HCE cells at clinic medication dosage. Timolol maleate should be carefully utilized in ophthalmic clinic.

关 键 词：人角膜内皮细胞 噻吗心安 细胞凋亡 DNA断片化 凋亡小体 

分 类 号：R775[医药卫生—眼科]