HPPH光动力学治疗鼠G422脑胶质瘤诱导肿瘤细胞凋亡的实验观察

Observe on Expression of the Tumor Cell Apoptosis Induced by HPPH-PDT in Treatment of G422 Glioma of Mice

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： The tumor cell apoptosis induced by novel photosentizer HPPH for photodynamic therapy of mice bearing G422 glioma was explored by FCM and electron microscope. The comparison of effect of HPPH-PDT and HpD-PDT was assessed. To ascertain the adequate dosage of HPPH of HPPH-PDT in treatment of G422 glioma of rats. Methods： The mice G422 glioma model in brain and subcutaneous axillary tumours was established. Mice suffering G422 glioma were randomly divided into seven groups： blank control, HPPH0.45mg/kg alone, 665nm laser alone, 630nm laser alone, HPPH -PDT （0.15mg/kg, 0.3mg/kg, 0.45mg/kg ） and HpD - PDT5mg/kg. Mice were injected intravenously with O.15,0.3,0.45mg/kg of HPPH utilizing 665nm diode laser doses of 204J/era2 at 200mW/cm2 delivered at 24 hours post drug injection for HPPH-PDT groups. For HPD-PDT group, 24 hours after intravenous injeetiort of HPD5mg/kg, 630rim diode laser doses of 240J/cm^2 at 200mW/cm^2 was given. Research on inhibition efficacy of tumors of Mice G422 Glioma by HPPH-based photodynamic therapy from apoptosis level of tumors cells was done by FCM and electron microscope.To ascertain the adequate dosage of HPPH of HPPH-PDT in treatment of G422 glioma of rats, the comparison of effect of HPPH-PDT and HPD-PDT was assessed. Results： No significant difference of apoptosis rate among blank control group,HPPH 0.45 mg/kg alone group, 665 nm laser alone group and 630 nm laser alone group. Compared with blank control group, apoptosis rate of HPPH-PDT groups and HPD-PDT group significantly increased, P〈0.01. Apoptosis rate of HPPH-PDT 0.3 mg/kg and 0.45 mg/kg group exceeded 0.15 mg/kg group and HPD-PDT group obviously, P〈0.01.0.45 mg/kg group exceeded 0.3 mg/kg group slightly, P〉 0.05. HPPH-PDT 0.3 mg/kg exceeded HPD-PDT group obviously, P〈0.01. Conclusion： HPPH-PDT can induce apoptosis of G422 glioma in correlation with dose of HPPH appropriate dose was 0.3mg/kg. From apoptosis level, the inhibition effect on G422 glioma by HPPH-PDT was better than HpD-PDT.