血卟啉光动力作用杀伤人胆管癌细胞的实验研究

Photodynamic therapy of hematoporphyrin derivative on human cholangiocarcinoma cell

目的研究血卟啉化衍生物(hematoporphyrin derivative,HPD)光动力作用(photodynamic therapy,PDT)对人胆管癌细胞QBC939的杀伤效应。方法以人胆管癌细胞系QBC939为研究对象,采用血卟啉化合物作为光敏剂,半导体激光治疗仪为光源。实验分为对照组(空白对照组、单纯光动力组、单纯光敏剂组)和光动力作用组,以系列浓度的血卟啉经不同强度光照后,用MTT法测定PDT对QBC939细胞的相对抑制率和筛选最佳PDT参数。结果光动力作用组的吸光度(OD值)与对照组间的差异具有统计学意义(P<0.01),随着HPD浓度的增加和光照强度的增大,光动力作用对细胞的相对抑制率逐渐增大。不同的光敏剂浓度和光照强度间组合能达到相同的抑制率,如以10mg/LHPD经5J/cm2光照强度或以4mg/LHPD经15J/cm2光照强度对QBC939均可以达到92%相对抑制率。在最佳光照能量密度条件下,设计三组功率-时间组合,组间没有显著差异。结论1)PDT对人胆管癌细胞QBC939具有明确的杀伤作用,其对细胞的抑制率具有显著的剂量效应关系。光敏剂浓度和光照强度间存在交互关系,从临床角度考虑,采用较低的光敏剂浓度经较大的光照强度照射是理想的PDT治疗方案。2)改变功率时间的组合不会影响光动力对胆管癌细胞杀伤作用,采用在光纤承受范围内的大功率短时间的照射方式可达到安全快捷的目的。

Objective To investigate the killing effect of photodynamic therapy （PDT） on human cholangiocarcinoma cell in vitro, and the dosage-effect relation of photo sensitizer and laser light in photody-namic therapy. Methods Human cholangiocarcinoma cells were cultured with serial concentrations of hemato-porphylin derivatives （HPD） followed by irradiation of different dosage of laser light, then MTT colorimetric assay was applied to measure the relative inhibitory, rate of PDT for the cells. Use SPSS software to find out the best parameter for killing effect of PDT. Observe the different killing effect of three groups of efficiency-time set-up. Results Significant difference in the relative inhibitory between the PDT group and control group was observed （P〈0.01）.The relative inhibitory rate of PDT for the cells elevated along with the increase in the concentration of sensitizer and dose of laser light. Relative inhibitory rate of the lower concentration and high laser light dose group will be the same as that of higher concentration HPD and lower laser light dose group. Conclusion PDT has significant killing effect on human cholangioncarcinoma cells, and its relative inhibitory rate appears to be correlated with the dose of sensitizer and laser light irritation. Low concentration and high laser light dose set-up is the effective and safe module of PDT. Higher laser light dose and shorter time irradiation will be the best pattern of PDT.