5-氨基酮戊酸-光动力疗法对HaCaT细胞增殖活性、细胞凋亡、原卟啉Ⅸ及活性氧的影响  

Effect of 5- aminolevulinic acid- photodynamic therapy on cell proliferation activity,cell apoptosis,protoporphyrin Ⅸ and reactive oxygen species of HaCaT keratinocytes

作　　者：田黎明[1] 王玮蓁[1] 刘茜茜[1] 潘凯[1] 彭圆[1] 

机构地区：[1]华中科技大学同济医学院附属武汉市第一医院皮肤科,湖北武汉430022

出　　处：《新乡医学院学报》2015年第11期981-984,共4页Journal of Xinxiang Medical University

基　　金：国家自然科学基金资助项目(编号:81101212;81574037);中国皮肤科医师协会-复旦张江光动力基金(编号:2012CDA-FDZJ01);湖北省自然科学基金面上项目(编号:2015CFB577);武汉市卫生局科学基金资助项目(编号:WX12B20)

摘　　要：目的观察5-氨基酮戊酸-光动力疗法(ALA-PDT)对Ha Ca T细胞增殖活性、细胞凋亡、原卟啉IX(Pp IX)及活性氧(ROS)的影响。方法体外培养Ha Ca T细胞,分成4组,对照组不加任何刺激因素,ALA组单用1 mmol·L-1ALA溶液加入Ha Ca T细胞培养基中避光孵育3 h,PDT组单用红光治疗仪照射Ha Ca T细胞培养液30 min,ALA-PDT组在加入1 mmol·L-1ALA溶液于Ha Ca T细胞培养基中避光孵育3 h后,用红光治疗仪照射上述培养液30 min。噻唑蓝(MTT)法检测细胞增殖活性,流式细胞仪检测细胞凋亡水平,荧光显微镜观察Pp IX的含量以及ROS检测试剂盒检测细胞内ROS的水平。结果对照组、ALA组、PDT组、ALA-PDT组细胞增殖活性分别为0.892±0.009、0.879±0.024、0.722±0.013、0.534±0.029;细胞凋亡率分别为(2.702±0.019)%、(2.697±0.031)%、(6.283±0.282)%、(32.601±1.897)%;ROS活性分别为(100.000±0.000)%、(101.333±3.786)%、(112.000±3.606)%、(135.000±16.000)%;PDT组细胞增殖活性、细胞凋亡率及ROS活性与对照组和ALA组比较差异有统计学意义(P<0.05);ALA-PDT组该3个指标与对照组、ALA组和PDT组比较,差异均有统计学意义(P<0.05)。对照组和ALA组细胞内Pp IX的荧光比率分别为(4.000±1.000)%、(32.000±2.646)%,2组比较差异有统计学意义(P<0.05)。结论 ALA诱导Ha Ca T细胞内Pp IX的累积,Pp IX在PDT作用下产生大量ROS,继而抑制Ha Ca T细胞增殖,导致细胞凋亡。Objective To observe the effect of 5-aminolevulinic acid-photodynamic therapy（ ALA-PDT） on cell proliferation activity,cell apoptosis,protoporphyrin IX（ Pp IX）,reactive oxygen species（ ROS） of human Ha Ca T keratinocytes.Methods Ha Ca T cells were cultured in vitro and divided into four groups. The control group receiving no treatment; the ALA group was given 1 mmol·L- 1ALA solution which was cultured in Ha Ca T cell culture medium under avoiding light for 3 h; in PDT group,the red light therapy instrument was taken to irradiate the Ha Ca T cell culture solution for 30 min; in ALA-PDT group,the red light therapy instrument was taken to irradiate the Ha Ca T cell culture solution for 30 min after 1 mmol·L- 1ALA solution was cultured in Ha Ca T cell culture medium under avoiding light for 3 h. The cell proliferation activity was determined with methylthiazolyldiphenyl-tetrazolium bromide（ MTT） method. The levels of apoptotic cells were measured by flow cytometry. Accumulation of Pp IX was observed using an fluorescence microscope. ROS was detected by ROS detection kit. Results In control group,ALA group,PDT group and ALA-PDT group,the cell proliferation activity was 0. 892 ± 0. 009,0. 879 ± 0. 024,0. 722 ± 0. 013,0. 534 ± 0. 029,respectively; the apoptosis rate was（ 2. 702 ± 0. 019） %,（ 2. 697 ±0. 031） %,（ 6. 283 ± 0. 282） %,（ 32. 601 ± 1. 897） %,respectively; the level of ROS was（ 100. 000 ± 0. 000） %,（ 101. 333 ±3. 786） %,（ 112. 000 ± 3. 606） %,（ 135. 000 ± 16. 000） %,respectively. There were statistic differences of cell proliferation activity,apoptosis rate and the level of ROS among the PDT group and control group,ALA group（ P〈0. 05）; there were statistic differences of cell proliferation activity,apoptosis rate and the level of ROS among the ALA-PDT group and control group,ALA group,PDT group（ P〈0. 05）. The percentage of Pp IX fluorescence of control group and ALA group was（ 4. 000 ±1. 000） %,（ 32. 000 ± 2. 646） %,respectively

关 键 词：5-氨基酮戊酸 光动力治疗 HaCaT细胞 细胞凋亡 原卟啉IX 活性氧 

分 类 号：R751[医药卫生—皮肤病学与性病学]