血管内皮细胞热损伤规律的初步探讨

Pilot Study on Thermal Damage Rules of Endothelial Cel

目的观察血管内皮细胞在不同温度条件下的坏死率和凋亡率,以期了解光动力治疗中的热效应规律。方法人血管静脉内皮细胞接种于96孔细胞培养板内。待细胞生长良好,数量达80%后,置于水浴锅加热,时间15 min。按不同的加热温度分为5组,分别为46℃组、49℃组、51℃组、55℃组和61℃组,每组接种10孔。Annexin-FITC染色后,用流式细胞仪检测细胞坏死率和凋亡率。结果 46℃组,人血管静脉内皮细胞的坏死率(10.09±1.44)%,凋亡率(17.57±2.42)%;49℃组,坏死率(9.68±1.53)%,凋亡率(12.55±1.60)%;51℃组,坏死率(36.97±2.78)%,凋亡率(21.29±2.87)%;55℃组,坏死率(47.04±3.06)%,凋亡率(16.19±1.43)%;61℃组,坏死率(61.99±2.40)%,凋亡率(20.30±4.65)%。细胞坏死率在51℃后呈逐步上升趋势,而凋亡率变化不大。经方差分析,细胞坏死率46℃组和49℃组比较,差异无显著意义(P>0.05),其余各组间差异有显著意义(P<0.01);而凋亡率49℃和51℃组比较,差异有显著意义(P<0.05),其余各组间差异无显著意义(P>0.05)。结论人血管静脉内皮细胞在加热15 min条件下,坏死率随温度升高而升高,而凋亡率变化不明显。

Objective Despite of being injured mainly by reactive oxygen species,endothelial cells may also be affected by the heat of thermal effect coming from tissue absorption of laser energy during photodynamic therapy（PDT） on port wine stains（PWSs）.This pilot study investigated the necrosis and apoptosis of endothelial cells in different temperatures,and tried to show the rules of thermal effect during PDT.Methods The human being endothelial cells were planted in 96 well plates.The plates were heated for 15 min in a water-bath when the wells were 80% full with cells of good conditions.The temperatures of the groups（each having 10 wells） were 46℃,49℃,51℃,55℃ and 61℃ respectively.The rates of necrosis and apoptosis were tested with flow cytometry after the dying with Annexin-FITC.Results The necrosis rate of endothelial cells was（10.09 ± 1.44） % and the apoptosis rate（17.57 ± 2.42） % at a temperature of46℃.The necrosis rate was（9.68 ± 1.53） % and the apoptosis rate（12.55 ± 1.60） % at a temperature of 49℃.The necrosis rate was（36.97 ± 2.78） % and the apoptosis rate（21.29 ± 2.87） % at a temperature of 51℃.The necrosis rate of endothelial cells was（47.04 ± 3.06） % and the apoptosis rate was（16.19 ± 1.43） % at a temperature of 55℃.The necrosis rate of endothelial cells was（61.99 ± 2.40） % and apoptosis rate（20.30 ± 4.65） % at a temperature of 61℃.The necrosis rate increased when the temperature climbed over 51℃,but the apoptosis rate rarely changed.The statistical analysis showed that there was no difference in necrosis rate between group 46℃ and group 49℃,and significant difference was found between other groups（P〈0.01）.The apoptosis rate was statistically different between group 49℃ and group 51℃（P〈0.05）,and no difference was observed between other groups.Conclusions The necrosis rate of human being endothelial cells increased along with the rise of temperature,and the apoptosis rate did not change obviously after a heating of 15 min.