摘要
目的探讨血卟啉衍生物介导的光动力学疗法对体外培养大鼠C6胶质瘤细胞的杀伤效应。方法①应用MTT法测定不同HpD浓度组(0,2,5,10,20,30及40mg/L)C6细胞PDT(628nm,20mW/cm2,照光5min)后的生存率;②采用光镜及电镜观察PDT后C6细胞的形态学变化;③DNA琼脂糖凝胶电泳检测各组样本的DNA条带;④TUNEL法检测各组样本PDT治疗后的细胞凋亡阳性率。结果①HpD浓度分别为2、5、10、20、30、40mg/L各组样本的细胞生存率分别为:110.31±6.25%、74.63%±6.57%、41.74%±4.16%、36.43%±2.14%、31.94%±2.25%及30.25%±2.88%;②PDT后C6细胞(HpD浓度>5mg/L时)出现细胞回缩等改变;透射电镜观察发现10mg/LHpD组C6细胞出现染色质边集、凋亡小体形成等改变;20mg/LHpD组C6细胞出现胞膜破裂崩解、细胞核肿胀等表现;③DNA琼脂糖凝胶电泳检测发现10mg/L及5mg/LHpD浓度组产生了明显的梯状条带;④TUNEL测定5mg/L及10mg/LHpD浓度组C6细胞凋亡阳性率分别为33%和63%;20mg/LHpD浓度组细胞凋亡阳性率为6%。结论当HpD浓度为2mg/L时PDT对C6胶质瘤细胞无杀伤效应;当HpD≥5mg/L时PDT治疗能够引起C6胶质瘤细胞出现细胞凋亡及坏死,较低浓度(5-10mg/L)时C6细胞以凋亡为主,较高浓度HpD(≥20mg/L)时C6细胞主要以坏死为主。
Objective To evaluate the effect of hepatoprophyrin derivative（HpD） mediated photodynamic therapy （PDT） on cultured C6 glioma cells. Methods （1）C6 glioma cells were irradiated at 628 nm, 20 mW/cm2 for 5 min afrer being incubated with different concentration of HpD （0, 2, 5, 10, 20, 30 and 40 mg/L）respectively, then cell survival rate was examined by MTT; （2）Inverted phase contrast microscope and transmitted eletron microscope （TEM） was used to examine the structural characteristics of C6 glioma cells of each group; （3）DNA agarose gel electrophoresis （AGE） was used to determine the DNA band of C6 glioma cells after HpD- PDT; （4）Apoptosis of C6 glioma cells were detected by TUNEL assay in differernt groups. Results （1）The cell survival rate of C6 cells in different HpD groups（2,5,10,20 and 40 mg/L）are 110.31 ± 6.25% .74.63± 6.57% .41.74±4.16% .36.43 ±2.14% .31.94±2.25% and 30.25 ± 2.88% respectively; （2）Morphological changes such as cell shrinkage were found after HpD-PDT; TEM observation found most C6 cells appeared a typical apoptotic changes in 10 mg/L group, While in 20 mg/L group, most of the C6 glioma cells appeared as necrotic; （3）DNA Agarose gel electrophoresis： Typical DNA ladder was seen in the 5 and 10 mg/L groups; （4）TUNEL detection showed： Some rather typical apoptotic positive cells were observed in 5 and 10 mg,/L grops with a proportionate of 33% and 63% respectively. Conclusions HpD-PDT can induce HpD concentration-related apoptosis and necrosis of C6 glioma cells, with a dominant apoptosis at low concentraion and necrosis at high concentraion; but in the low HpD concentration （2 mg/L）group, PDT didn＇t kill the C6 cells but seemed to promote cell growth.
出处
《国际神经病学神经外科学杂志》
2008年第1期12-17,共6页
Journal of International Neurology and Neurosurgery
基金
国家自然科学基金资助项目:30670506
国家十一五科技支撑计划(2007DAI05D08)
