正交设计优化光动力治疗宫颈癌的作用参数

目的:采用正交实验设计,探讨姜黄素作为光敏剂介导的光动力治疗诱导宫颈癌Me180细胞凋亡的最佳作用参数。方法:选定光敏剂姜黄素的药物浓度、激光剂量、照射时间、孵育时间四个因素,分别按姜黄素浓度2.5、5、10μmol/L,激光剂量50、100、200J/cm~2,照射时间90、180、360s,孵育时间12、24、36h。每因素3个水平设计正交设计表,按正交设计表进行姜黄素作为光敏剂介导的光动力治疗。治疗后细胞继续培养,MTT法测定细胞生存率。通过双因素方差分析三个水平的姜黄素剂量和激光剂量的估计边缘平均值,寻找最佳治疗参数。结果:对细胞凋亡的影响因素:姜黄素剂量孵育时间照射时间激光剂量;各因素水平影响:姜黄素浓度:10μmol/L5μmol/L2.5μmol/L;激光剂量:200J/cm~2100J/cm~250J/cm~2;照射时间:360s180s90s;孵育时间:24h12h36h。双因素方差分析表明,姜黄素浓度2.5μmol/L、5μmol/L、10μmol/L三组之间两两比较,差异均有统计学意义(P0.05);100J/cm~2、200J/cm~2与50J/cm~2组间比较,差异均有统计学意义(P0.05),但100J/cm~2与200J/cm~2组间比较差异无统计学意义。结论:姜黄素介导光动力治疗诱导宫颈癌细胞凋亡最佳参数组合为姜黄素浓度5μmol/L,激光剂量100J/cm~2,照射时间180s,孵育时间24h。

Optimization of photodynamic therapy induced apoptosis of cervical cancer cells by or-thogonal design

Objective:To search the optimum parameters of curcumin as a photosensiti-zer of photodynamic therapy induced apoptosis of human cervical carcinoma Me180 cells by or-thogonal design. Methods:The selected photosensitizer curcumin concentration,laser dosage,ir-radiation time, incubation time four factors, according to the concentration of curcumin 2. 5μmol/L,5μmol/L,10μmol/L,50J/cm2 ,100J/cm2 ,200J/cm2 laser dose,irradiation time:90s, 180s,360s,incubation time:12h,24h,36h. 3 levels of each factor design and orthogonal design table,photodynamic therapy of curcumin by orthogonal design table as a photosensitizer,after treatment of cells,cell survival rate was measured by MTT method. The average marginal value of three levels of curcumin and laser dosage were estimated by two factor analysis of variance. Results:The order of influence factors of cell apoptosis induced by PDT is:curcumin dose >in-cubation time > the laser irradiation time >laser dosage. The level of various factors:Curcumin concentration:10μmol/L>5μmol/L>2. 5μmol/L;200J/cm2>100J/cm2>50J/cm2 laser dose;irradiation time:360s>180s>90s;incubation time:24h>12h>36h. Double factor variance analy-sis showed that curcumin concentration of 2. 5μmol/L,5μmol/L,10μmol/L among the three groups were statistically significant differences(P<0. 05);100J/cm2,200J/cm2 and 50J/cm2 group,there was a statistically significant difference (P<0. 05),100J/cm2 and 200J/cm2 but no statistically significant difference between the groups. Conclusions:Curcumin mediated photo-dynamic therapy induced apoptosis of cervical cancer cells with the best combination of curcu-min concentration of 5μmol/L,laser dose of 100J/cm2 ,irradiation time of 180s,incubation time of 24h.