5-氨基酮戊酸光动力疗法对体外培养的增生性瘢痕成纤维细胞生长和增殖影响的初步研究

Effects of 5-Aminolaevulinic aacid photodynamic therapy on cell growth and proliferation of fibroblast in vitro of hypertrophic scar

目的:探讨5-氨基酮戊酸光动力疗法(ALA-PDT)对体外培养的增生性瘢痕组织的成纤维细胞生长和增殖的影响,观察细胞的形态学及超微结构的变化,并探讨可能的机制。方法:采用四甲基偶氮唑盐(MTT)比色法检测ALA,激光照射对体外培养的成纤维细胞增殖的影响;1mmol/L的ALA接受在不同的时间激光照射对成纤维细胞增殖的影响;在光镜和电镜下观察ALA-PDT对细胞形态和超微结构的影响;用分光光度法检测ALA-PDT对细胞氧自由基含量、超氧化物歧化酶活性的影响。结果:单一的激光照射对成纤维细胞的生长没有影响;1、10、102和103mmol/L的ALA对细胞生长有一定的抑制作用(P<0.05;1mmol/L的ALA接受20min、40min和60min激光照射后细胞的增殖明显下降(P<0.05),呈时间依赖关系;ALA-PDT作用于细胞后,细胞体积变大,正常的细胞连接消失,细胞内的线粒体减少、体积增大,内质网呈空泡化;ALA-PDT作用于细胞后,细胞ROS含量升高,SOD活性降低(P<0.05)。结论:ALA-PDT对体外成纤维细胞具有明显的损伤作用,对细胞的增殖和生长有一定的抑制作用;其作用机制可能与ALA-PDT对细胞的氧化损伤有关。

Objective To investigate the effect of 5-aminolaevulinic acid photodynamic therapy（ALA- PDT） on growth and proliferation of fibroblast of hypertrophic scar in vitro and its possible mechanism. Methods The effect of ALA-PDT on fibroblast activities was determined by M-IT assay. The morphological and ultrastructure changes of fibroblasts exposed to ALA-PDT were observed with invert microscope and transmission electron microscope. The content of reactive oxygen species （ROS） in cells and activities of superoxide dismutase （SOD） in cells were determined by spectrophotometric method. Results The proliferation of fibroblasts was inhibited by ALA-PDT. After ALA-PDT, the fibroblasts displayed typical morphological and ultrastructure changes of cell damage. After ALA-PDT, the content of ROS significantly increased, while the activities of SOD decreased（both P〈0.05）. Conclusion ALA-PDT can inhibit the proliferation and growth of the skin fibroblasts. Furthermore, oxidative damage may be involved the injury of ALA-PDT to cells.