论文部分内容阅读
目的:通过对鼠G422脑胶质瘤脑及腋部皮下移植瘤模型的HPPH光动力学治疗,从电镜、FCM测定细胞凋亡率,观察各剂量组疗效,并与对照组及HpD-PDT作对比,寻找HPPH-PDT治疗鼠G422脑胶质瘤合适的HPPH剂量。方法:建立鼠G422脑胶质瘤脑及腋部皮下移植瘤模型,设立空白对照组、单注射HPPH 0.45 mg/kg组、单注射HpD组、单665 nm激光照射组、单630 nm激光照射组、HPPH-PDT各组(0.15、0.3、0.45 mg/kg组)、HpD-PDT 5 mg/kg组。单注射HPPH组、HPPH-PDT组和单注射HpD组、HpD-PDT组自尾静脉注入光敏剂,24 h后以波长665 nm的半导体激光照射HPPH-PDT组和单665 nm激光组肿瘤,功率密度200 mW/cm~2,每光斑照射17 min,能量密度为204 J/cm~2;以波长630 nm的半导体激光照射HpD-PDT组及单630nm激光照射组肿瘤,功率密度200 mW/cm~2,每光斑照射20 min,能量密度为240 J/cm~2。于PDT后9 d处死鼠,取肿瘤、肿瘤边缘、正常脑组织,作电镜、FCM细胞凋亡检查。结果:以流式细胞仪及电镜观察肿瘤细胞凋亡,显示鼠G422脑胶质瘤脑及皮下移植瘤HPPH-PDT各剂量组、HpD-PDT组与空白、单注药和单照光三组对照组比较,肿瘤细胞凋亡率明显升高,P<0.01,差别有统计学意义。HPPH-PDT各组细胞凋亡率高于HpD-PDT组,0.3 mg/kg、0.45 mg/kgHPPH-PDT组的细胞凋亡率明显高于0.15 mg/kgHPPH-PDT组,HpD-PDT组,P<0.01,差别有统计学意义,0.45 mg/kg HPPH-PDT组细胞凋亡率稍高于0.3 mg/kgHPPH-PDT组,P>0.05,差别无统计学意义。故HPPH0.3 mg/kg是适宜的HPPH-PDT光敏剂剂量。HPPH-PDT0.3 mg/kg组和HpD-PDT组比较,肿瘤细胞凋亡率明显升高,P<0.01,差别有统计学意义。故由肿瘤细胞凋亡率和电镜分析,HPPH-PDT能诱导鼠G422脑胶质瘤细胞凋亡,凋亡率与HPPH剂量相关,适宜的剂量为0.3 mg/kg;HPPH-PDT诱导鼠G422脑胶质瘤细胞凋亡的作用较HpD-PDT强。结论:从电镜、FCM观察HPPH-PDT对鼠G422脑胶质瘤生长有抑制作用,能诱导肿瘤细胞凋亡及死亡。凋亡率与HPPH剂量有关,适宜的剂量为0.3 mg/kg;HPPH-PDT诱导鼠G422脑胶质瘤细胞凋亡的作用较HpD-PDT强。
OBJECTIVE: To observe the effect of each dose group by HPPH photodynamic therapy on mouse brain and axillary subcutaneously implanted tumor model of G422 glioma. The apoptosis rate was observed by electron microscopy and FCM, and compared with the control group and HpD-PDT In contrast, look for HPPH-PDT treatment of mouse G422 glioma appropriate HPPH dose. Methods: The brain and axillary subcutaneous xenografts model of G422 glioma was established. A blank control group was established. Single injection of HPPH 0.45 mg / kg group, single injection of HpD group, single 665 nm laser irradiation group and single 630 nm laser irradiation group , HPPH-PDT groups (0.15,0.3,0.45 mg / kg group) and HpD-PDT 5 mg / kg group. Single injection of HPPH group, HPPH-PDT group and single injection of HpD group, HpD-PDT group were injected photosensitizer from the tail vein, 24 h after the wavelength of 665 nm semiconductor laser irradiation HPPH-PDT group and single 665 nm laser group tumor, power At a density of 200 mW / cm ~ 2, the energy density was 204 J / cm ~ 2 at 17 min for each spot, and the tumor was irradiated with HpD-PDT group and single 630 nm laser at a wavelength of 630 nm. The power density was 200 mW / cm ~ 2, irradiating 20 min per spot, the energy density is 240 J / cm ~ 2. Rats were sacrificed 9 days after PDT, and the tumors, tumor margins and normal brain tissues were taken for electron microscopy and FCM. Results: Apoptosis of tumor cells was observed by flow cytometry and electron microscopy. The results showed that HPPH-PDT in each dose group and HpD-PDT group were significantly different from blank, single-injection and single-light control groups The apoptosis rate of tumor cells was significantly increased, P <0.01, the difference was statistically significant. The apoptosis rate in HPPH-PDT groups was higher than that in HpD-PDT group. The apoptosis rate in HPP-PDT group was significantly higher than that in HPP-PDT group (0.3 mg / kg, 0.45 mg / <0.01, the difference was statistically significant, 0.45 mg / kg HPPH-PDT group apoptosis rate slightly higher than 0.3 mg / kgHPPH-PDT group, P> 0.05, the difference was not statistically significant. Therefore, HPPH0.3 mg / kg is suitable HPPH-PDT photosensitizer dose. HPPH-PDT0.3 mg / kg group and HpD-PDT group, the apoptosis rate of tumor cells was significantly increased, P <0.01, the difference was statistically significant. Therefore, HPPH-PDT can induce the apoptosis of G422 glioma cells by the apoptosis rate of tumor cells and electron microscopy. The apoptosis rate was related to HPPH dose, the suitable dose was 0.3 mg / kg; HPPH-PDT induced G422 brain Glioma cell apoptosis than HpD-PDT stronger. Conclusion: The observation of electron microscopy and FCM showed that HPPH-PDT could inhibit the growth of G422 glioma cells and induce the apoptosis and death of tumor cells. The apoptosis rate was related to the dosage of HPPH, and the suitable dosage was 0.3 mg / kg. The apoptosis of G422 glioma cells induced by HPPH-PDT was stronger than that of HpD-PDT.