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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 study the apoptosis of vascular endothelial cells (RPE) and retinal pigment epithelium (RPE) induced by photodynamic therapy (PDT) in vitro. Methods Vascular endothelial cells and human RPE cells cultured in vitro were incubated with the verteporfin photosensitizer at a concentration of 1.0μg / ml. According to the incubation time, each cell was divided into 6 groups of 0, 5, 15, 30, 60,120 min group. After the above incubation, the maximum absorption wavelength of the photosensitizer (689 nm, diode laser with a power density of 600 mW / cm2) was used, respectively, with an energy density of 2.4 J / cm2 and a light time of 83 s. Flow cytometry 3h after each group of apoptosis ratio, the experiment was repeated 3 times. Results The apoptosis rates of vascular endothelial cells in 6 groups 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. The percentage of apoptosis in each group 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. The percentages of apoptosis of both cells increased with the incubation time of cells and photosensitizers. The two cells were in the same There was no significant difference in the percentage of apoptosis in the time group by t test (P> 0.05). Conclusion PDT can induce rapid apoptosis of RPE cells and vascular endothelial cells. Under the same in vitro environment, PDT has no obvious selectivity to the apoptosis induced by the two kinds of cells.