摘要
目的探讨去铁胺对δ-氨基酮戊酸（ALA）诱导的皮肤成纤维细胞光动力学效应的影响。方法体外培养人皮肤成纤维细胞，分为空白对照组、ALA组、去铁胺组及ALA＋去铁胺组。37℃下避光孵育3h后，检测细胞内原卟啉Ⅸ（PpⅨ）水平、细胞增殖情况（吸光度A值）和细胞存活率。对检测出PpⅨ的细胞组予以632．8nm波长He—Ne激光照射，流式细胞术测定细胞死亡率与凋亡率。结果He—Ne激光照射前，各组细胞的吸光度A值和存活率比较，差异均无统计学意义（P〉0．05）；空白对照组与去铁胺组的细胞皆未检测出PpⅨ，而ALA组与ALA＋去铁胺组的细胞内PpⅨ水平分别为（0．47±0．04）μg／L和（0．85±0．08）μg／L，2组差异有统计学意义（P〈0．05）。He-Ne激光照射后，ALA组和ALA＋去铁胺组的细胞凋亡率分别为14．1％和21．5％，2组差异有统计学意义（P〈0．05）；细胞坏死率分别为2．7％和3．0％，2组差异有统计学意义（P〈0．05）。结论ALA和去铁胺都不影响皮肤成纤维细胞的基本生物学特征，去铁胺可促进ALA诱导的细胞内PpⅨ生成并增强光动力学效应。
Objective To investigate the effect of deferoxamine （DFO） on the photodynamic reaction in human skin fibroblasts co-cultured with δ-aminolevulinic acid （ALA）. Methods Human skin fibroblasts were isolated from juvenile foreskin tissue and co-cultured in vitro with nothing （control group） , 2 mmol/L ALA, 0.1 mmol/L DFO or ALA ＋ DFO. Three hours later, the protoporphyrin IX （PpIX） levels in the fibroblasts were determined by high performance liquid chromatography with fluorescent detection （HPLC-FLD）. Laser scanning confocal microscopy （LSCM） was used to observe the strength of fluorescence in the fibroblasts with the excitation of a 488 nm laser. The proliferation （ absorbance ratio） and survival rate of cells were detected by MTT colorimetric methods and trypan blue staining respectively. The cellular PpIX positive groups were irradiated with a 632.8 nm He-Ne laser. Flow cytometry was used to calculate cellular apoptosis and necrosis. Results There was no significant difference between the cell absorbance ratio or survival rates of the four groups before He-Ne laser iradiation. No cellular PpIX was found in the control or DFO groups, while the cellular PpIX concentrations in the ALA and ALA ＋ DFO groups were （0.47 ± 0.04 ） μg/L and （0.85 ± 0.08） μg/L respectively. After He-Ne laser irradiation, in the ALA and ALA ＋ DFO groups the cellular apoptosis rates were 14. 1% and 21.5% respectively, and the cellular necrosis rates were 2.7% and 3.0% respectively. Conclusions Neither ALA nor DFO affected the basic biological features of skin fibroblasts, but DFO could enhance the PpIX production and subsequent photodynamic reactions in human skin fibroblasts co-cultured with ALA.
出处
《中华物理医学与康复杂志》
CAS
CSCD
北大核心
2010年第2期114-117,共4页
Chinese Journal of Physical Medicine and Rehabilitation
