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Biological effects observed 5 - aminolevulinic acid photodynamic therapy (5-aminolevulinic acid photodynamic therapy, ALA-PDT) in peripheral blood mononuclear cells (peripheral blood mononuclear cells, PBMCs) of healthy people and patients with psoriasis vitro explore ALA-PDT in the treatment of psoriasis immune mechanism, feasibility and optimal treatment parameters to provide a theoretical basis for the clinical application of this therapy. l, isolation and culture of psoriasis patients and healthy human peripheral blood mononuclear cells. MTT colorimetric detection of healthy human PBMCs were treated with 0.1 mmol / L, 0.6 mmol / L, 1.2 mmol / L, 1.8 mmol / L, 3.6 mmol / L of ALA incubated in the dark 0.5h, 1h, 3h, 5hPDT survival and filter out the best drug concentration and optimum Incubate time. 3, Annexin V / PI double staining combined with flow cytometry healthy human PBMCs were treated with 0.1 mmol / L, 0.6 mmol / L, 1.2 mmol / L, 1.8mmol / L to 3.6mmol / L of ALA incubated in the dark 0.5h 1h, 3h apoptosis situation 5hPDT, the further validate optimal drug concentration and the optimal dark time. 4, transmission electron microscopy healthy human PBMCs by 0.6mmol / L of ALA dark incubation 1h PDT Ultrastructural Changes. 5, RT-PCR to detect expression of Th1, Th2 cytokines affect ALA-PDT of psoriasis patients and healthy human PBMCs. 1, MTT method showed that healthy human PBMCs were treated with 0.1 mmol / L, 0.6 mmol / L, 1.2mmol / L and 1.8mmol / L 3.6mmol/LALA incubated in the dark 0.5h, 1h, 3h and 5h after PDT results to 0.6mmol / L ALA Incubate 1h ALA-PDT group lowest cell viability, compared with the blank control group, the difference was statistically significant (P lt; 0.05). 2, compared with the blank control group, healthy human PBMCs 0.6mmol / L of ALA Incubate 1h PDT obvious apoptosis after Annexin V / PI staining combined with flow cytometry showed the group of early apoptosis was 8.5 ± 0.5% (P lt; 0.05). Observed under transmission electron microscope to healthy human PBMCs by 0.6mmol / L of ALA dark incubated for 1h PDT showed typical apoptotic changes, no apoptotic cells in the blank control group. 4, RT-PCR assay showed that patients with psoriasis vulgaris PBMCs, IFN-γ, TNF-αmRNA expression levels compared to healthy individuals significantly increased expression levels decreased significantly by 0.6 mmol / L ALA dark incubated for 1h PDT , in which the expression of IFN-ymRNA reduced to normal levels; significant decrease in IL-4 mRNA expression in PBMCs than healthy people, significantly increased after ALA-PDT, but did not reach the level of healthy people; patients the PBMCs the IL-10mRNA of expression than healthy people slightly reduce elevated after ALA-PDT, but there was no statistical significance. 1, healthy human PBMCs by 0.6mmol / L ALA Incubate 1h after PDT for dark time optimal drug concentration and the most appropriate. 2, healthy human PBMCs by 0.6mmol / L of ALA Incubate 1h after PDT can significantly promote apoptosis of peripheral blood mononuclear cells. 3 psoriasis patients with peripheral blood mononuclear cells after ALA-PDT response by Thl advantage to shift the direction of Th1/Th2 balance.
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