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为了揭示常用青光眼药物贝特舒(betaxolol)对人角膜上皮(HCEP)细胞的毒性及其作用机理,使用不同质量浓度贝特舒对体外培养HCEP细胞进行了处理,并利用倒置显微镜跟踪观察了细胞的生长和形态,用吖啶橙/溴化乙锭(AO/EB)荧光双染色法检测了质膜的通透性,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。发现在0.175 00~2.800 00 g/L的质量浓度范围内,贝特舒对HCEP细胞具有显著的毒性,并随着质量浓度的提高和处理时间的延长而逐渐增大,处理24 h即可使大部分细胞皱缩死亡;进一步的研究结果发现,质量浓度为0.175 00~2.800 00 g/L的贝特舒能显著提高HCEP细胞的质膜通透性,并使其出现DNA断片化、染色质凝缩和形成凋亡小体等凋亡细胞的结构变化。可见,在贝特舒0.175 00~2.800 00 g/L的质量浓度范围内对HCEP细胞具有显著的毒性,并具有质量浓度和时间依赖性,其毒性作用的发挥是通过诱导细胞凋亡实现的,在眼科临床应用中毒副作用极大。
In order to reveal the toxicity of betaxolol, a commonly used glaucoma drug, on human corneal epithelial cells (HCEP), HCEP cells cultured in vitro were treated with different concentrations of bettose, and the cells were observed by inverted microscopy The morphology and morphology of the plasma membrane were examined by acridine orange / ethidium bromide (AO / EB) fluorescence double staining. DNA fragmentation was detected by agarose gel electrophoresis. The cells were detected by transmission electron microscopy Ultrastructure. It was found that Bertucx was significantly toxic to HCEP cells in the mass concentration range of 0.17500 ~ 2.800 00 g / L and gradually increased with the increase of mass concentration and the prolongation of treatment time. Treatment with 24 h Most of the cells shrink to death; further study found that the concentration of 0.17500 ~ 2.800 00 g / L of Beitou can significantly increase the plasma membrane permeability of HCEP cells, and make it appear DNA fragmentation, chromatin Condensation and formation of apoptotic bodies and other apoptotic cells. It can be seen that in the mass concentration range of 0.17500 ~ 2.800 00 g / L, Berthell has significant toxicity to HCEP cells in a concentration-dependent and time-dependent manner, and its toxicity is exerted through induction of apoptosis, Ophthalmology in clinical toxic side effects.