摘要
目的研究体外光动力疗法(PDT)诱导的血管内皮细胞和视网膜色素上皮(RPE)细胞凋亡情况。方法将体外培养的血管内皮细胞和人RPE细胞与血药浓度(1.0μg/ml)相当的光敏剂verteporfin共同孵育,根据孵育时间的不同,每种细胞分成6组0、5、15、30、60、120min组。孵育至上述时间后,分别用光敏剂最大吸收波长光照(689nm、功率密度为600mW/cm2的二极管激光),能量密度为2.4J/cm2,光照时间为83s。用流式细胞仪检测3h后各组细胞凋亡比例,实验重复进行3次。结果PDT后3h,血管内皮细胞在6组的凋亡比例分别为0.01±0.01、0.25±0.02、0.32±0.02、0.41±0.04、0.49±0.03和0.61±0.02;RPE细胞各组的凋亡比例分别为0.02±0.01、0.22±0.01、0.31±0.02、0.38±0.03、0.47±0.05和0.58±0.03;两种细胞的凋亡比例均随细胞与光敏剂孵育时间的延长而增加;两种细胞在相同时间组的凋亡比例经t检验差异均无统计学意义(P>0.05)。结论PDT可致RPE细胞和血管内皮细胞快速凋亡,在相同的体外环境下,PDT对两种细胞所诱导的凋亡反应无明显选择性。
Objective To detect the apoptosis of vascular endothelial ceils and retinal pigment epithelial （RPE） ceils in vitro induced by verteporfin-photodynamic therapy. Methods Cultured vascular endothelial cells and human RPE cells were incubated with verteporfin at a concentration of 1.0 μg/ml which was equivalent to the initial plasma level of verteporfin in clinical therapy. Each kind of cells were divided into 6 groups according to different time of incubation： 0, 5, 15, 30, 60, and 120 minutes group. After incubated, the cells were illuminated by the laser light with the maximum wavelength of absorption of verteporfin （wavelength： 689 nm, power density： 600 mW/cm^2） with the power of 2. 4 J/cm^2 for 83 seconds. The percentage of cellular apoptosis was measured by flow cytometry 3 hours after PDT, and the measurement was repeated thrice. Results The proportion of cellular apoptosis 3 hours after PDT were 0. 01±0. 01, 0. 25±0. 02, 0. 32±0. 02, 0. 41±0. 04, 0. 49±0. 03 and 0. 61±0. 02, respectively in 0-120 minutes group of vascular endothelial cells;and 0.02 ± 0.01, 0.22± 0.01, 0.31 ±0.02, 0.38±0.03, 0.47±0.05 and 0.58±0.03 respectively in 0-120 minutes group of RPE cells. The proportion of cellular apoptosis of both kinds of the cells increased as the incubation time was prolonged. There was no significant difference of the percentage of cellular apoptosis between the accordant time groups in the two kinds of cells （P〉0.05）. Conclusions Cellular apoptosis can be quickly induced by verteporfin-PDT both in human vascular endothelial cells and RPE cells; under the same condition in vitro, PDT has no obvious selection for the apoptosis of the two kinds of cells.
出处
《中华眼底病杂志》
CAS
CSCD
北大核心
2006年第4期253-255,共3页
Chinese Journal of Ocular Fundus Diseases
基金
广东省重点科技项目(99M04702G)
