目的:研究ALA、HPD及ALA联合HPD光动力学治疗对C6胶质瘤细胞细胞内钙离子变化。方法:以不同剂量ALA(20μg/ml、40μg/ml、60μg/ml、80μg/ml、160μg/ml)、HPD(1μg/ml、2μg/ml、3μg/ml、4μg/ml、5μg/ml)及不同剂量组合ALA联合HPD光动力学治疗C6胶质瘤细胞,应用Fluo-3AM为细胞内钙离子荧光指示剂,以激光共聚焦显微镜测定对照组及光动力学组照光后培养24小时活细胞细胞内钙离子含量,及测定单激光及对照组、光动力学治疗组照光前36秒及照光20分钟至停照光后20-100分钟细胞内钙离子浓度的动态变化,每12秒测定一次。结果:空白对照组、单激光、单加光敏剂组活C6胶质瘤细胞显示钙离子荧光值,在120分钟内变化较小。而光动力学治疗组,初照光5-15分钟内荧光明显慢慢增强,以后就逐渐下降,光敏药剂量大的初5-10分钟内荧光亮度上升很快,以后下降也快,可下降为0,剂量越大变化越明显,且光敏剂ALA较HPD更明显。二光敏剂联合组以上反应更明显,HPD1-2μg/ml联合ALA40-60μg/ml与HPD 5μg/ml组动态图相似。结论:通过本实验可推测光动力学作用后细胞内钙离子超载可能在细胞凋亡及死亡中发挥重要作用。二光敏剂联合可提高疗效,降低HPD光敏剂用量,减少皮肤光毒付反应,HPD1-2μg/ml联合ALA40-60μg/ml是较合适的剂量。 Objective: To study the changes of intracellular calcium in C6 glioma cells treated by ALA, HPD and ALA combined with HPD photodynamic therapy. Methods: HPD (1μg / ml, 2μg / ml, 3μg / ml, 4μg / ml and 5μg / ml) were treated with different doses of ALA (20μg / ml, 40μg / ml, 60μg / ml, 80μg / ml, 160μg / And different doses of combination of ALA combined with HPD photodynamic therapy of C6 glioma cells, the use of Fluo-3AM as an indicator of intracellular calcium fluorescence confocal laser microscopy of the control group and photodynamic therapy group light 24 hours after incubation of living cells Intracellular calcium content, and the determination of single laser and control group, photodynamic therapy group 36 seconds before light and light 20 minutes to stop light 20-100 minutes after intracellular calcium concentration changes measured every 12 seconds. Results: C6 glioma cells in blank control group, single laser and photosensitizer alone showed fluorescence intensity of calcium ion, which changed little in 120 minutes. The photodynamic therapy group, the initial light within 5-15 minutes, the fluorescence was slowly increased gradually decreased later, the first 5-10 minutes of the photosensitizer dose fluorescent brightness rose rapidly, after the decline is also fast, can be reduced to 0, the greater the dose changes more obvious, and photosensitizer ALA more obvious than the HPD. The combination of two photosensitizers more reaction was more obvious, HPD1-2μg / ml combined with ALA40-60μg / ml and HPD5μg / ml group dynamic picture is similar. CONCLUSION: It is inferred from this experiment that intracellular calcium overload may play an important role in apoptosis and death after photodynamic therapy. The combination of two photosensitizers can improve curative effect, reduce the dosage of HPD photosensitizer, and reduce the skin phototoxicity. HPD1-2μg / ml combined with ALA40-60μg / ml is more appropriate dosage.