5-氨基酮戊酸介导的光动力对人增生性瘢痕成纤维细胞的影响

Effects of 5-aminolaevulinic acid-mediated photodynamic therapy on cultured dermal fibroblasts from hypertrophic scar

目的探讨5-氨基酮戊酸介导的光动力对人增生性瘢痕中成纤维细胞的影响。方法选取人增生性瘢痕增生期组织5例，体外培养其中的成纤维细胞。取培养后的第3、4代成纤维细胞，添加5-氨基酮戊酸培养后应用激光共聚焦显微镜检测其代谢产物原卟啉Ix在细胞内的积聚，并在5-氨基酮戊酸作用5h后给予635nm波长的红光照射，照射功率密度10mW／em。，能量密度0．5～8．0J／em。。24h后，应用CCK-8试剂盒分析5-氨基酮戊酸介导的光动力对成纤维细胞的杀伤作用。结果5-氨基酮戊酸作用4h后，成纤维细胞中有原卟啉IX的积聚；作用5h后，原卟啉Ix的积聚达到高峰，此时给予激光照射，成纤维细胞的成活率降低，并与照射强度呈量效依赖关系。结论5．氨基酮戊酸介导的光动力能够杀伤增生性瘢痕中的成纤维细胞，是一种治疗增生性瘢痕的新方法。

Objective To investigate the effects of 5-aminolaevulinic acid-mediated photodynamic therapy （5-ALA-PDT） on cultured dermal fibroblasts from hypertrophic scar. Methods Hypertrophic scar samples were obtained from 5 patients who underwent surgery. Dermal fibroblasts were isolated and then cultured in vitro. The 3rd or 4th generation fibroblasts were used and co-cultured with different concentrations of 5-ALA. Intracellular protoporphyrin IX （PpIX） was measured under laser confocal microscope. The cells treated with 5-ALA for 5 hours were irradiated by red laser （635 nm wavelength） at a power density of 10 mW/cm= with ener- gy density from 0.5 to 8.0 J/cm2. The cell survival was measured by CCK-8 Kit at 24 hours. Results Intra-cellular PpIX accumulation was observed at 4 hours after 5-ALA-treatment. The peak level of PpIX accumula- tion was achieved at 5 hours after 1 mmol/L 5-ALA treatment. Viable cells were decreased after laser irradiation at a dose dependent manner with energy density. Conclusion Dermal fibroblasts from hypertrophic scar could efficiently accumulate PpIX by exogenous 5-ALA administration and be killed by laser irradiation. It is a new method to prevent clinical hyperplastic scar from formation.