盐酸奥布卡因对人角膜内皮细胞影响作用的实验研究  

Experimental studies on the effect of oxybuprocaine hydrochloride on human corneal endothelial cells

作　　者：樊廷俊[1] 温茜[1] 于苗苗[1] 葛源[1] 苗莹[1] 王德平[1] 

机构地区：[1]中国海洋大学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》2012年第8期1442-1446,共5页International Eye Science

基　　金：国家高技术研究发展863计划( No.2001AA625050)~~

摘　　要：目的:揭示眼科局部麻醉剂盐酸奥布卡因(oxybuprocaine hydrochloride,OBPC-HCl)对体外培养人角膜内皮(HCE)细胞的影响作用,为眼科临床安全用药提供实验依据。方法:用不同浓度OBPC-HCl处理体外培养的HCE细胞,在倒置显微镜下观察细胞的生长和形态变化,用吖啶橙/溴化乙锭(AO/EB)荧光双染色法检测质膜的通透性,用琼脂糖凝胶电泳法检测DNA的断片化,用透射电镜检测细胞的超微结构。结果:OBPC-HCl在62.5mg/L～4g/L的浓度范围内均能不同程度地引起HCE细胞出现细胞皱缩、胞内空泡化、质膜通透性增大、染色质凝缩、凋亡小体和DNA断片化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4g/LOBPC-HCl对HCE细胞的凋亡诱导作用最大,处理1h后HCE细胞的凋亡率已高达100%。结论:OBPC-HCl在62.5mg/L～4g/L的浓度范围内能显著诱导HCE细胞凋亡,在眼科临床应用中对HCE细胞的毒副作用极大。AIM： To examine the effect of oxybuprocaine hydrochloride （OBPC-HCI） on in vitro cultured human corneal endothelial （HCE） cells and provide foundations for clinical safety use of OBPC-HCI. METHODS： After treated with OBPC-HCI at different concentrations, the growth situation and morphology of in vitro cultured HCE cells were checked under an inverted light microscope, membrane permeability was detected by acridine orange/ethidium bromide （AO/EB） double- fluorescent staining. DNA fragmentations and ultrastructure of HCE cells were observed by DNA agarose gel electrophoresis and transmission electron microscopy （TEM）, respectively. RESULTS： In vitro cultured HCE cells treated with OBPC- HCI at a concentration between 62. 5mg/L and 4g/L showed typical characteristics of apoptosis, including cell shrinkage, intercellular vacuolation, membrane permeability increasing, chromatin condensation, apoptotic body appearance and DNA fragmentation. The apoptosis- inducing effect of OBPC-HCI was in dose- and time- dependent manners. The greatest apoptosis- inducing effect of OBPC-HCI on HCE cells was found at the clinic concentration of 4g/L, and the apoptotic rate of HCE cells reached 100% after treated for 1 hour. CONCLUSION： OBPC-HCI at a concentration of 62.5mg/L- 4g/L has an obvious apoptosis-inducing effect on HCE cells, and has huge toxic side effects on HCE cells during its clinic ophthalmic usage.

关 键 词：人角膜内皮细胞 盐酸奥布卡因 细胞凋亡 DNA断片化 凋亡小体 

分 类 号：R614[医药卫生—麻醉学]