新型靶向光敏剂诱导Hep-2细胞光氧化行为的研究  

Research on the Photooxidative Action in Hep-2 Cells Induced by A Novel Targeting Photosensitizer

作　　者：李东红[1] 李鹏熙[1] 蒋宗林[2] 郭林峰[2] 

机构地区：[1]创伤,烧伤,复合伤国家重点实验室,第三军医大学大坪医院,重庆400042 [2]西华师范大学,四川南充637002

出　　处：《肿瘤学杂志》2013年第7期539-543,共5页Journal of Chinese Oncology

基　　金：国家自然科学基金资助项目(21072227)

摘　　要：[目的]研究新型光敏剂Ⅰ诱导Hep-2细胞的光氧化行为。[方法]利用MTT法检测光敏剂Ⅰ对Hep-2细胞的细胞毒性;采用活性氧特异性探针H2DCFDA通过激光共聚焦成像观察Hep-2细胞中活性氧的生成;通过测定超氧化物歧化酶(SOD)、谷胱甘肽(GSH)和丙二醛(MDA)水平及乳酸脱氢酶(LDH)渗漏检测观察Hep-2细胞的氧化应激反应。[结果]无光照时,光敏剂Ⅰ对Hep-2细胞的毒性为零,但光照后可明显抑制该细胞的生长,且其光毒性随光照剂量的增加而加强(r=-0.962,P=0.001)。光动力治疗后,细胞内DCFDA的荧光强度逐渐增强,在4h时达到高峰,随后又逐渐降低;细胞内SOD和GSH水平逐渐降低,3h后分别降低42.5%(P<0.01)和35.0%(P<0.01),而MDA含量却随时间延长逐渐增加,3h后增加54%(P<0.01)。LDH的渗出与光照剂量呈正相关(r=0.966,P=0.007)。[结论]新型光敏剂Ⅰ可有效光诱导Hep-2细胞死亡,而细胞内氧化应激反应可能是其光诱导Hep-2细胞死亡的重要作用机制。[Purpose] To investigate the photooxidative action of Hep-2 cells induced by a novel photosensitizer Ⅰ. [Methods] The cytotoxicity of photosensitizer Ⅰ against Hep-2 cells was measured by MTT assay. The formation of active oxygen in the Hep-2 cells induced by photodynamic therapy（PDT） were observed by using an active oxygen specificity probe H 2 DCFDA through confocal laser scanning microscopy. The intracellular oxidative stress was investigated by determination of the levels of superoxide dismutase（SOD）,glutathione （GSH） and malondialdehyde （MDA） in Hep-2 cells and the effusion of lactic dehydrogenase（LDH）. [Results] No toxicity of photosensitizer Ⅰ on Hep-2 cells was observed when irradiation was not applied. Hep-2 cells were inhibited after PDT, and the photocytotoxicity of photosensitizer Ⅰ increased with the augmentation of irradiation（r=-0.962, P=0.001）. After PDT,the fluorescence intensity of DCFDA in cells increased gradually,and reached the peak at 4h,then decreased gradually; the levels of SOD and GSH decreased gradually,with de- creased 42.5% （P0.01）and 35.0% （P0.01）respectively 3h after PDT; but the level of MDA in- creased with the prolongation of time,and increased 54% 3h after PDT （P0.01）. The effusion of LDH was positively correlated with irradiation dose （r=0.966,P=0.007）. [Conclusion] PDT mediated by photosensitizer Ⅰ can effectively induce the death of Hep-2 cells,and the oxidative stress in cells maybe the main mechanism.

关 键 词：靶向性光敏剂Ⅰ HEP-2细胞 光动力治疗 氧化应激反应 

分 类 号：R73-3[医药卫生—肿瘤]