光动力作用对人血管内皮细胞分泌TNF-α的影响

Effect of photodynamic treatment on the secretion of tumor necrosis factor oL by human vascular endothelial cells

目的 观察血卟啉单甲醚光动力疗法（HMME-PDT）处理人血管内皮细胞（EC-304）后分泌TNF-α浓度变化,以探讨细胞因子与光动力疗法治疗鲜红斑痣后出现炎症反应的关系.方法将EC-304接种于6孔培养板,实验分为HMME-PDT实验组,HMME对照组,激光对照组,空白对照组.实验组在光动力疗法处理后12、24、48h分别取上清液,对照组于换液后相同时间点分别取上清液.ELISA法测量4组TNF-α的分泌量.结果每一组TNF-α在不同时间点的含量存在差异（F=62.276,P〈0.01）,4组之间TNF-α的含量在每个相同的时间点不完全相等（F=11.538,P〈0.01）,经多重比较,实验组与3组对照组之间TNF-α的含量在每个相同的时间点均有差异（P〈0.01）,3组对照组之间在每个相同的时间点的含量差异无统计学意义（P〉0.05）.结论 HMME-PDT可促进EC-304分泌TNF-α.

Objective To measure the changes in levels of tumor necrosis factor α（TNF-α） secreted by human endothelial cells EC-304 after hematoporphyrin monomethyl ether-photodynamic treatment （HMME-PDT）,and to explore the relationship between cytokines and inflammation initiation after management of nevus flammeus with photodynamic therapy.Methods EC-304 cells were cultured in 6-well plates,and classified into 4 groups：HMME-PDT group pretreated with HMME followed by irradiation with laser,HMME control group treated with HMME only,laser control group irradiated with laser only,and blank control group without any treatment.Culture supematants of EC-304 cells were collected from HMME-PDT group at 12,24 and 48 hours after the irradiation,and from the other three groups at the same time points.The supernatant TNF-α level was measured with enzyme linked immunosorbent assay （ELISA）.Results The difference was statistically significant in the supernatant TNF-α level between different time points in each group （F=62.276.P〈0.01） and between the 4 groups at each time point （F=11.538,P〈0.01）.Multiple comparison analysis showed that HMME-PDT group differed significantly from the other 3 control groups in the supernatant TNF-αlevel at each time point （all P〈0.01）,while no significant difierence was observed among the other three control groups at any time point （all P〉0.05）.Conclusion HMME-PDT promotes the secretion of TNF-α by EC-304 cells.