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

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。

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

Objective: The tumor cell apoptosis induced by novel photosentizer HPPH for photodynamic therapy of mice bearing G422glioma 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 inbrain and subcutaneous axillary tumours was established. Mice suffering G422 glioma were randomly divided into seven groups: blankcontrol、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 0.15、0.3、0.45mg/kg of HPPH utilizing 665nm diode laser doses of 204J/cm2 at 200mW/cm2 delivered at 24 hours post drug injection for HPPH -PDT groups. For HPD -PDT group, 24 hours after intravenousinjection of HPD5mg/kg, 630nm diode laser doses of 240J/cm2 at 200mW/cm2 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 electronmicroscope.To ascertain the adequate dosage of HPPH of HPPH-PDT in treatment of G422 glioma of rats, the comparison of effectof 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 rateof 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 G422glioma in correlation with dose of HPPH appropriate dose was 0.3mg/kg. From apoptosis level, the inhibition effect on G422 glioma byHPPH-PDT was better than HpD-PDT.