中华皮肤科杂志 ›› 2009, Vol. 42 ›› Issue (9): 628-631.

• 论著 • 上一篇    下一篇

0.1%二甲基亚砜促进δ-氨基酮戊酸诱导HaCaT细胞光动力学效应

夏育民1,刘小明2,王首帆3,熊腊元4,徐世正3   

  1. 1. 武汉大学人民医院皮肤科江珊收转13659892065
    2. 武汉大学人民医院
    3. 武汉大学人民医院皮肤科
    4. 武汉湖北医科大学附属第一医院皮肤科 430060
  • 收稿日期:2008-08-26 修回日期:2009-05-19 出版日期:2009-09-15 发布日期:2012-03-21
  • 通讯作者: 夏育民 E-mail:xiayumin1202@163.com

δ-Aminolevulinic acid-based photodynamic reaction in HaCaT cells is enhanced by a low concentration of dimethyl sulfoxide

  • Received:2008-08-26 Revised:2009-05-19 Online:2009-09-15 Published:2012-03-21
  • Contact: Yu-min XIA E-mail:xiayumin1202@163.com

PDF (PC)

294

赞

摘要： 目的 探讨0.1%二甲基亚砜（DMSO）对δ-氨基酮戊酸诱导HaCaT细胞光动力学效应的影响。方法 将HaCaT细胞分成实验组（0.1% DMSO）和对照组（未加DMSO），加入2 mmol/L δ-氨基酮戊酸并于37 ℃避光孵育3 h。采用高效液相色谱荧光法（HPLC-FLD）检测胞内原卟啉Ⅸ（PpⅨ）水平，用激光共聚焦显微镜（LSCM）观察胞内PpⅨ荧光强度，同时检测细胞增殖（噻唑蓝法测定吸光度A值）与存活率；以632.8 nm波长激光照射HaCaT细胞后继续培养12 h，通过流式细胞仪检测细胞凋亡率与坏死率。结果 HPLC-FLD检测实验组和对照组HaCaT细胞质内PpⅨ水平分别为（0.57 ± 0.05） μg/L和（0.44 ± 0.04） μg/L（t = 2.79，P < 0.05），LSCM观察实验组胞内荧光强度高于对照组，吸光度A值分别为0.54 ± 0.06、0.51 ± 0.07（t = 1.51，P > 0.05），细胞存活率分别是（96.18 ± 2.25）%，（94.64 ± 2.40）%（χ2 = 1.84，P > 0.05）。照光后，实验组和对照组的细胞凋亡率分别为2.2%，1.5%（χ2 = 4.05，P < 0.05），细胞坏死率分别为8.9%和0.1%（χ2 = 8.23，P < 0.05）。结论 0.1% DMSO可促进δ-氨基酮戊酸所致的HaCaT细胞光动力学效应。

关键词: 角蛋白细胞, 二甲亚砜, 氨基酮戊酸, 血红素, 光化学疗法

Abstract: Objective To investigate the effect of 0.1% dimethyl sulfoxide （DMSO） on photodynamic reaction in a human keratinocyte cell line, HaCaT, induced by 5-aminolevulinic acid. Methods HaCaT cells were cultured in vitro with or without the presence of 0.1% DMSO at 37 ℃ and stimulated by δ-aminolevulinic acid （ALA） for 3 hours with light free. Then, cellular protoporphyrin Ⅸ （PpⅨ） concentration in, fluorescence intensity in, proliferation and survival rates of HaCaT cells were determined by high performance liquid chromatography with fluorescence detection （HPLC-FLD）, laser scanning confocal microscopy （LSCM）, MTT colorimetric method and trypan blue staining, respectively. Also, a portion of HaCaT cells were treated with ALA-based photodynamic therapy （ALA-PDT） and irradiated by 632.8-nm laser, and 12 hours later, cellular apoptosis and necrosis were detected by flow cytometry with annexinⅤ/PI staining. Results Increased concentrations of PpⅨ were found in DMSO-pretreated HaCaT cells compared with untreated cells （0.57 ± 0.05 μg/L vs 0.44 ± 0.04 μg/L, P < 0.05）, while no statistical difference was observed in fluorescence intensity expressed as the absorbance at 570nm （0.54 ± 0.06 vs 0.51 ± 0.07, t = 1.51, P > 0.05） or cell survival rate （（96.18 ± 2.25）% vs（94.64 ± 2.40）%, χ2 = 1.84, P > 0.05） between DMSO-pretreated and untreated cells. After ALA-PDT, the apoptosis and necrosis rate were significantly increased in DMSO-treated HaCaT cells compared with untreated HaCaT cells （2.2% vs 1.5%, χ2 = 4.05, P < 0.05; 8.9% vs 0.1%, χ2 = 8.23, P < 0.05）. Conclusion Low concentration （0.1%） of DMSO could enhance the effect of ALA-PDT on HaCaT cells.