摘要
目的:通过研究ALA-PDT对体外培养的PBMCs的生物效应,探讨ALA-PDT对银屑病的免疫功能的影响、治疗的可行性及优化参数。方法:将健康人及银屑病患者新鲜外周血分别进行PBMCs分离与培养,分设空白对照组、单纯ALA组、单纯照光组以及不同ALA浓度与不同用药孵育时间组进行PDT。Annexin V/PI双染法、MTT法检测PBMCs存活率,以确定ALA最佳药物浓度和最佳用药孵育时间;TEM观察PBMCs细胞超微结构的变化;RT-PCR检测PBMCs表达Th1、Th2相关细胞因子的水平。结果:0.6 mmol/LALA避光孵育1 h后PDT组PBMCs存活率最低(P<0.05);ALA-PDT后PBMCs结构呈现典型凋亡改变;银屑病患者PBMCs的IFN-γ、TNF-αmRNA表达量与健康人相比显著升高,ALA-PDT后表达量显著下降、IL-4 mRNA表达量较健康人显著降低,经ALA-PDT后显著升高(P<0.05);IL-10mRNA表达量较健康人略有减低,经ALA-PDT后有所升高,但无统计学意义。结论:0.6 mmol/LALA避光作孵育1 h后PDT可明显促进PBMCs凋亡,银屑病患者PBMCs经ALA-PDT后由Th1优势应答向Th1/Th2平衡方向转变。
Objective：In the paper,we observed the effects of ALA-PDT on PBMCs,thus to explore the immune effect,treatment feasibility and optimized parameter of ALA-PDT on psoriasis.Methods：we isolated peripheral blood mononuclear cells from healthy people and psoriatic patients respectively.The experiment was divided into control group,ALA only group,light only group,and ALA-PDT groups including different ALA concentrations and different periods of incubation time.Annexin V/PI double-staining combined with flow cytometry（FCM） was used to detect the apoptosis of PBMCs to further verify the selected concentration and incubation time.Transmission electron microscope（TEM） was used to observe the ultra-structure changes of PBMCs after ALA-PDT. RT-PCR was used to detect the mRNA expression of T helper1/T helper2（Th1/Th2）-type cytokines of PBMCs.Results：The ALA-PDT group incubated with 0.6 mmol/L ALA for 1 hour showed the highest PBMCs apoptosis rate（P0.05）.Apoptotic PBMCs were observed by TEM after treatment with ALA-PDT,but none was observed in control group.RT-PCR showed that ALA-PDT can clearly decrease the mRNA expression of IFN-γand TNF-αin psoriatic PBMCs,which was significantly higher than that in healthy control before treatment.ALA-PDT markedly increased the mRNA expression of IL-4 in psoriatic PBMCs,which was lower than that in healthy control before treatment.IL-10 mRNA in psoriatic PBMCs was a little bit lower than that in healthy control,and raised some after ALA-PDT.Conclusions：ALA-PDT could markedly promote cell apoptosis of PBMCs at 0.6mmol/L concentration and 1 hour incubation time.Abnormal Th1 immune responses of psoriatic PBMCs was downregulated after ALA-PDT,meanwhile Th2 immune responses was upregulated.
出处
《应用激光》
CSCD
北大核心
2011年第6期518-523,共6页
Applied Laser
