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-PDT5 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, 24h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/cm2每光斑照射 17 min, 能量密度为 204 J/cm2; 以波长 630 nm的半导体激光照射 HpD-PDT组及单 630 nm激光照射组肿瘤, 功率密, 度 200 mW/cm2每光斑照射 20 min, 能量密度为 240 J/cm2。于 PDT后 9d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 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.