CD147 siRNA转染对银屑病患者外周血T淋巴细胞CD147的表达及细胞增殖、活化的影响

Study on psoriatic peripheral blood T-lymphocytes： effects of CD147-targeting small interfering RNA on the expression of CD147 and on cell proliferation and activation

目的探讨转染CD147siRNA对寻常性银屑病患者外周血T淋巴细胞中CD147的表达及细胞增殖、活化的影响。方法将化学合成的CD147siRNA通过电转染至寻常性银屑病患者外周血T淋巴细胞中，采用半定量RT—PCR方法检测细胞在转染96h内CD147mRNA的表达变化。采用噻唑蓝法检测转染CD147siRNA对T淋巴细胞增殖的影响。采用流式细胞仪检测转染CD147siRNA对T淋巴细胞活化标记分子CD25表达的影响。结果与未转染组比较，转染CD147siRNA24h后，T淋巴细胞的CD147mRNA表达水平显著下降（P〈0．05），至转染后48h降低最为明显（P〈0．01），而转染后96h恢复至接近转染前水平（P〉o．05）；转染CD147siRNA24h、48h、72h后，T淋巴细胞的增殖水平显著下降，抑制率分别为（44．5±3．13）％、（50．7±3．5）％、（53．98±4．15）％（_P值均〈0．01），CD25的表达也明显降低（P值均〈0．01）。结论CD147分子与银屑病外周血T淋巴细胞的增殖、活化能力相关，可能是一种新的银屑病治疗靶分子。

To investigate the effects of small interfering RNA （siRNA） on the expression of CD147 on peripheral blood T-lymphocytes from patients with psoriasis vulgaris, and its effect on the proliferation and activation of these cells. Methods Peripheral blood monouclear cells （PBMC） were obtained from 10 patients with psoriasis vulgaris, and T lymphocytes were isolated. CD147 siRNA was chemically synthesized, then electroporated into the peripheral T-lymphocytes. Untransfected cells, blank-transfected cells and unspecifically transfected cells served as the control. After 24-, 48-, 72- and 96-hour incubation, RT-PCR was used to detect the mRNA expression of CD147 in these cells. MTT assay and flow cytometry were utilized to assess the proliferation of these cellas, and the expression of CD25 at 24, 48, and 72 hours after the transfection. Results Compared with untransfected cells, the mRNA expression of CD147 declined significantly in CD147 siRNA-transfected cells at 24 hours （P 〈 0.05）, reached to the minimum at 48 hours （P 〈 0.01 ）; there was no significant difference in the expression of CD147 between the two groups of cells at 96 hours after the transfection （P 〉 0.05）. There was a decrease of cell proliferation level by （44.5 ± 3.13 ）%, （50.7 ± 3.5）% and （53.98 ± 4.15）% in CD147 siRNA-transfected cells 24, 48 and 72 hours following the transfection, respectively; the corresponding decrease in blank-transfected cells was （37.28 ± 3.56）%, （33.73 ± 3.29）%, and （28.80 ± 1.49）%, respectively, and that in unspecifically transfected cells, （31.29 ± 2.46）%, （36.1 ± 2.62）% and （32.08± 2.78）%, respectively. A significant decrease was observed in the proliferation of CD147 siRNA-transfected cells compared with that of blank-transfected cells and unspecifically transfected cells at these three time points （P 〈 0.05, 0.01, 0.01 respectively）. The expression rate of CD25 at 24, 48 and 72 hours was （47.23 ± 3.65）%, （31.50 ±4.22）% and （23.05 ±4.15）%, respectively, on CD147 siRNA-transfected cells, and, （80.2 ±4.8）%, （81.6 ±3.35）% and （83.5 ± 4.1）%, respectively, on untransfected cells; the differences between the two groups at the three time points were statistically significant （all P 〈 0.01 ）. Conclusion CD147 is correlated with cell proliferation and activation of peripheral T lymphocytes from patients with psoriasis vulgaris, and may serve as a new treatment target for psoriasis.