文章摘要
戴维德1,曾晶2,李晓松2,刘凡光2,顾瑛2.光动力效应致靶细胞线粒体损伤的机制研究[J].山西医科大学学报,2009,40(1):13~15
光动力效应致靶细胞线粒体损伤的机制研究
Mechanism of photodynamic effect induced mitochondrial damage of target cell
  
DOI:
中文关键词: 光动力效应  线粒体损伤  荧光探针标记术
英文关键词: photodynamic effect  mitochondrion damage  fluorescence probe labeling technique
基金项目:国家自然科学基金资助项目(60078021)
作者单位
戴维德1,曾晶2,李晓松2,刘凡光2,顾瑛2 1卫生部北京医院超声科北京1007302解放军总医院激光科 
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中文摘要:
      目的〓 探讨光动力效应对靶细胞线粒体损伤的发生机制。 〓方法〓 先将血卟啉单甲醚与ECV304细胞共同孵育12 h,采用荧光探针标记技术和细胞器-细胞荧光强度比值法对细胞内HMME进行亚细胞定位和定量分析。应用荧光显微镜汞灯照射以激发光敏剂产生光动力效应,并加入ROS探针H2DCF DA检测单线态氧的产生。分别在光照前后采集线粒体及DCF的荧光图像并进行荧光强度的定量分析。 〓结果〓 细胞总荧光光强在光照后较光照前降低26.4%;以细胞器光照前的平均荧光光强比值为基准,则光照后线粒体降低了31.1%。线粒体区域在单纯光照和有HMME存在情况下均检测到DCF荧光的增强。 〓结论〓 光动力效应会引起线粒体内光敏剂发生一定程度的光漂白作用,表明线粒体内产生了一定数量的单线态氧,后者导致线粒体损伤。
英文摘要:
      Objective〓 To explore a mechanism of target cell mitochondrial damage induced by photodynamic effect. 〓Methods〓 ECV304 cells were incubated with hematoporphyrin monomethyl ether for 12 h.Fluorescence probe labeling techniques and organelle cell fluorescence intensity ratio analysis were adopted to investigate the subcellular localization of HMME.Mercury lamp of fluorescence microscopy was used as light source to make photosensitizers excitated and produce photodynamic effects.Fluorescence probe of ROS H2DCF DA was used to detect production of singlet oxygen.Fluorescence images of mitochondrion and DCF were recorded before and after irradiation. 〓Results〓 Total fluorescence intensity of cells was decreased by 26.4% after irradiation.With mitochondrial average fluorescence intensity ratio before irradiation as the standard,the total fluorescence intensity of mitochondrion was decreased by 31.1% after irradiation.Enhancement of fluorescence of DCF was detected in mitochondrion with HMME or with single light after irradiation. 〓Conclusion〓 Photodynamic effects may cause a certain degree of photobleaching of photosensitizers in mitochondrion,which indicates that a certain quantity of singlet oxygen exist in mitochondrion to induce its damage.
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