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目的:研究ALA、HPD及ALA联合HPD光动力学治疗鼠G422胶质瘤肿瘤细胞内钙离子变化。方法:以不同剂量ALA(20、40、60、80、160mg/kg、)、HPD(1、2、3、4、5mg/kg)及不同剂量组合ALA联合HPD光动力学治疗鼠G422胶质瘤肿瘤,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立即、一天、三天肿瘤活细胞细胞内钙离子含量,并同时测定单激光、单用光敏剂、及空白对照组鼠肿瘤细胞内钙离子表达。结果:空白对照组、单激光、单加光敏剂组鼠G422胶质瘤肿瘤显示3天内钙离子荧光值维持在较恒定值,光动力组在治疗后立即组可见钙离子值明显高于未作光动力学治疗的对照组,且与光敏剂剂量增加有关,光动力学治疗后一天及三天组肿瘤细胞内钙离子值明显低于立即组及未作光动力学治疗组,且与光敏剂剂量增加有关。ALA-PDT组较HPD-PDT组更明显。联合治疗组HPD1-2mg/kg联合LA40-60mg/kg光动力学治疗组肿瘤细胞内钙离子值接近HPD 5mg/kg光动力学治疗组。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,减少皮肤光毒副反应,HPD1-2 mg/kg联合ALA40-60 mg/kg是较合适的剂量。
Objective: To study the intracellular calcium changes in G422 glioma tumor cells treated by ALA, HPD and ALA combined with HPD photodynamic therapy. Methods: G422 glomerulus was treated with different doses of ALA (20, 40, 60, 80, 160mg / kg), HPD (1,2,3,4,5mg / kg) and different doses of ALA combined with HPD photodynamic therapy Fluo-3AM was used as intracellular calcium fluorescence indicator. The contents of intracellular calcium in the viable cells in the control group and the photodynamic group were measured immediately after laser irradiation by laser scanning confocal microscopy. Laser, single photosensitizer, and blank control group. Results: G422 gliomas in blank control group, single laser group and photosensitizer alone group showed that the calcium fluorescence value maintained at a constant value within 3 days. The photodynamic value of calcium group in the photodynamic therapy group immediately after treatment was significantly higher than that of the control group Photodynamic therapy of the control group, and photosensitizer dose-related, photodynamic therapy one day and three days after the tumor cell calcium was significantly lower than the immediate group and no photodynamic therapy group, and the photosensitizer Increased dose. The ALA-PDT group was more obvious than the HPD-PDT group. In combination therapy group, HPD1-2mg / kg and LA40-60mg / kg photodynamic therapy group intracellular calcium ion close to HPD 5mg / kg photodynamic therapy group. 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 the curative effect, reduce the dosage of HPD photosensitizer, and reduce the photo-toxicity of the skin. HPD 1-2 mg / kg combined with ALA 40-60 mg / kg is more appropriate dosage.