李东红,李鹏熙,蒋宗林.新型靶向光敏剂诱导Hep-2细胞光氧化行为的研究[J].肿瘤学杂志,2013,19(7):539-543.
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
投稿时间:2012-09-17  
DOI:10.11735/j.issn.1671-170X.2013.07.B008
中文关键词:  靶向性光敏剂Ⅰ  Hep-2细胞  光动力治疗  氧化应激反应
英文关键词:targeting photosensitizer Ⅰ  Hep-2 cells  photodynamic therapy  oxidative stress
基金项目:国家自然科学基金资助项目(21072227)
作者单位
李东红 创伤、烧伤、复合伤国家重点实验室第三军医大学大坪医院 
李鹏熙 创伤、烧伤、复合伤国家重点实验室第三军医大学大坪医院 
蒋宗林 西华师范大学 
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中文摘要:
      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。
英文摘要:
      Abstract:[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 H2DCFDA 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 decreased 42.5%(P<0.01)and 35.0%(P<0.01)respectively 3h after PDT; but the level of MDA increased with the prolongation of time,and increased 54% 3h after PDT(P<0.01). 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.
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