雷公藤内酯醇对原发免疫性血小板减少症患者浆细胞样树突细胞功能及成熟的影响

Regulations of function and maturation of plasmacytoid dendritic cells from primary immune throm- bocytopenia patients by triptolide

目的探讨雷公藤内酯醇对原发免疫性血小板减少症（ITP）患者浆细胞样树突细胞（pDC）功能及成熟的影响。方法取22例初治ITP患者（初治组）、22例糖皮质激素治疗后完全缓解（CR）ITP患者（CR组）及22名健康志愿者（正常对照组）外周静脉血，分离单个核细胞，用流式细胞术分选pDC，分别加入0、5、10、30μg/L的雷公藤内酯醇共孵育，24h后收集上清液，用ELISA法检测IFN-α、IL-6、TNF—α水平；5d后收集细胞，用流式细胞术检测髓样树突细胞（mDC）CD11c、CD80、CD86表达，光镜下观察mDC的形态，电镜观察mDC的超微结构。结果加入10μg/L雷公藤内酯醇后初治组IFN—α、IL-6、TNF—α水平分别为（451．32±85．77）、（105．68±23．85）、（135．78±30．62）ng／L，CR组分别为（391．71±72．49）、（84．73±17．77）、（108．16±23．21）ng／L，正常对照组分别为（335．51±67．54）、（73．62±21．82）、（95．58±32．85）ng／L，初治组及CR组高于正常对照组（P〈0．05），初治组高于CR组（P〈0．05），并呈雷公藤内酯醇浓度依赖性（P〈0．05）；初治组及CR组mDC表型CD11c、CD80、CD86阳性率高于正常对照组（P〈0．05），初治组高于CR组（P〈0．05），加入雷公藤内酯醇后mDC表型阳性率呈浓度依赖性降低（P〈0．05）。加入雷公藤内酯醇后mDC较对照组突起变少、变短，形态上不成熟。结论雷公藤内酯醇能够减弱ITP患者pDC的功能，并抑制其向mDC的分化和成熟。

Objective To explore the mechanism of immunomodulatory activity of triptolide on pri- mary immune thrombocytopenia（ITP） patients-derived plasmacytoid dendritic cells （pDCs）. Methods pDCs in peripheral blood of ITP patients before therapy （ group 1 ） , ITP patients in complete response （ ITP-CR, group 2） and healthy donors （ group 3 ） were sorted by flow cytometry, then incubated with triptolide at 0, 5, 10 or 30 μg/L. After 24 hours, we collected the supernatants and then detected the concentrations of IFN-α, IL-6 and TNF-α using ELISA. After 5 days, the cultured cells were collected and CDlle, CDSO and CD86 expressions of myeloid dendritic cells （mDCs） were analyzed by flow cytometry, the morphology of mDC was observed by light microscope and electron microscope. Results After incubation with triptolide at 10 μg/L, the levels of IFN-α, IL-6 and TNF-α in group 1 [ （451.32 ±85.77） ng/L, （ 105.68 ±23.85 ） ng/L and （ 135.78 ± 30.62） ng/L] and group 2 [ （ 391.71 ±72.49 ） ng/L, （ 84.73 ±17.77） ng/L and （ 108.16 ± 23.21 ）ng/L] were significantly higher than those in group 3 [ （335.51 ± 67.54） ng/L, （73.62 ± 21.82 ） ng/L and （95.58 ± 32.85 ） ng/Ll （ all P 〈 0.05 ） ; the levels of IFN-α, IL-6 and TNF-α in group 1 were significantly higher than those in group 2 （all P 〈 0.05 ） in a dose-dependent manner （P 〈 0.05）. CD1 lc, CDS0 and CD86 expressions of mDC in groupl and group2 were significantly higher than those in group 3 （ all P 〈 0.05）; CDIIc, CD80 and CD86 expressions of mDC in group 1 were significantly higher than those in group 2 （ all P 〈 0. 05 ） also in a dose-dependant manner （ all P 〈 0. 05 ）. Triptolide could inhibit pDCs from differentiation into mDCs, the latter displayed more immature morphology than untreated-pDCs. Conclusion Triptolide could decrease the immune function of pDCs from ITP, inhibit pDCs from differentiation and maturation.