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目的:通过细胞学水平研究确定ALA配合小剂量HPD激光光动力疗法促使肿细胞凋亡及死亡的光敏药用药剂量及与两者单独使用的差别。方法:以不同剂量的ALA(20μg/ml、40μg/ml、60μg/ml、80μg/ml、160μg/ml)、HPD(1μg/ml、2μg/ml、3μg/ml、4μg/ml、5μg/ml)、两者联合与C6胶质瘤瘤细胞一起培养,随后以630nm半导体激光200mW/cm2,照射肿瘤细胞,照射20分钟,及与不用光敏剂单照激光、单用光敏剂不照光、不用光敏剂不照激光空白对照组比较。以流式细胞仪及倒置显微镜、电镜观察促使肿瘤细胞凋亡或死亡的差别,寻找促使肿瘤细胞凋亡或死亡的最小合适剂量。结果:ALA40μg/ml配合HPD2.5μg/ml与C6胶质瘤细胞同时培养,630半导体激光200mW/cm2,照射20分钟组从倒置显微镜及电镜形态学观察及流式细胞仪测定是能达到较佳的细胞凋亡的最小光敏剂剂量。单纯ALA-PDT组中能达到小鼠C6胶质瘤细胞明显凋亡的最小剂量是ALA(80μg/ml),单纯HPD-PDT组中能达到小鼠C6胶质瘤细胞明显凋亡最小剂量是HPD(5μg/ml)。结论:ALA40mg/kg配合HPD2.5mg/kg,630半导体激光200mW/cm2,照射20分钟是能达到小鼠C6胶质瘤细胞明显凋亡的最小光敏剂剂量。
OBJECTIVE: To determine the dose of photosensitizer that ALA combined with low-dose HPD laser photodynamic therapy can induce the apoptosis and death of swollen cells by cytology level and the difference with the two alone. Methods: The effects of different doses of ALA (20μg / ml, 40μg / ml, 60μg / ml, 80μg / ml, 160μg / ml), HPD (1μg / ml, 2μg / ml, 3μg / ml, 4μg / ml, 5μg / ml ), The two combined with C6 glioma cells cultured together, followed by 630nm semiconductor laser 200mW / cm2, irradiation of tumor cells, irradiation for 20 minutes, and with no photosensitizer single-shot laser, single photosensitizer does not light, no photosensitive Comparison of blank control group with no dose of laser. Observed by flow cytometry and inverted microscope, electron microscopy to promote the difference of apoptosis or death of tumor cells to find the smallest suitable dose to promote tumor cell apoptosis or death. Results: ALA 40μg / ml with HPD 2.5μg / ml and C6 glioma cells cultured at the same time, 630 semiconductor laser 200mW / cm2, irradiation 20 minutes from the inverted microscope and electron microscopy morphological observation and flow cytometry is better The minimum dose of photosensitizer for apoptosis. The minimal dose of ALA (80μg / ml) which could reach the obvious apoptosis of mouse C6 glioma cells in the ALA-PDT group was the lowest, which could reach the minimum level of obvious apoptosis in C6 glioma cells in the HPD-PDT group HPD (5 μg / ml). Conclusion: ALA40mg / kg with HPD2.5mg / kg, 630 semiconductor laser 200mW / cm2, irradiation for 20 minutes is the minimum photosensitizer dose that can achieve obvious apoptosis in mouse C6 glioma cells.