摘要
目的探讨δ-氨基酮戊酸（ALA）诱导的光动力学反应强度对皮肤成纤维细胞主要生物学特征的影响。方法体外分离并培养人皮肤成纤维细胞，加入终浓度为0．5～5．0mmol／L的ALA，37℃避光孵育3h后，632．8nm波长激光照射，通过MTT比色法、流式细胞仪检测细胞的增殖水平（A值）、死亡率，并用ELISA和碱性水解法测定细胞合成基质金属蛋白酶（MMP-1、2、3）、胶原蛋白（羟脯氨酸含量）的水平。结果随着ALA浓度上升，皮肤成纤维细胞的A值由0．45±0．05逐渐下降至0．32±0．04，而死亡率由6．4％±2．0％逐渐上升至29．6％±2．2％。同时，细胞合成MMP的水平先期呈逐渐增强，最后则逐渐减退；而胶原蛋白含量在初期呈下降趋势，在后期出现反弹。结论适当强度的光动力学反应可以促进皮肤成纤维细胞分泌MMP并抑制胶原蛋白合成，更强的光动力学反应则具有相反作用。
Objective To investigate the effect of 8-aminolevulinic acid （ALA）-induced photodynamic reaction on biological characteristics of skin fibroblasts. Methods Human skin fibroblasts were isolated from the foreskin of children and cultured in the presence of 0.5 to 5.0 mmol/L ALA; three hours later, they were irradiated with 632.8 nm laser followed by additional 12-hour culture. Then, MTT assay and flow cytometry were used to measure the proliferation and apoptosis of cells. The levels of matrix metalloproteinase （MMP）- 1, -2 and -3 and hydroxyproline in the culture supernatant of irradiated cells were determined by avidinbiotin complex-based ELISA and alkaline hydrolysis-based method, respectively. Results With the rise of ALA concentration from 0.5 to 5.0 mmol/L, the proliferation level of fibroblasts expressed as the absorbance at 570 nm dropped from 0.45 ± 0.05 to 0.32 ± 0.04, and cell death rate increased from 6.4% ± 2.0% to 29.6% ± 2.2%. Meanwhile, the contents of MMP-1, -2 and -3 increased at the early stage, but decreased at the late stage, whereas the hydroxyproline level showed an inverse tendency during the increase of ALA concentration from 0.5 to 5.0 mmol/L. Conclusions Proper intensity of photodynamic reaction induced by ALA could enhance the secretion of MMP and inhibit the synthesis of collagen by skin fibroblasts, however, high concentration of ALA may exert an inverse effect.
出处
《中华皮肤科杂志》
CAS
CSCD
北大核心
2010年第1期28-31,共4页
Chinese Journal of Dermatology
