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目的:研究大剂量地塞米松对免疫性血小板减少性紫癜(ITP)患者浆样树突状细胞(pDCs)功能及Toll样受体9表达的影响。方法:15例初诊的ITP患者给予地塞米松40mg/d,连用4d。采用免疫磁珠分离法体外分离15例正常对照及13例治疗有效患者治疗前后外周血中浆细胞样树突状细胞(pDCs),用CPG-ODN 2216刺激外周血pDCs并与之共培养24h,采用酶标记免疫吸附(ELISA)方法检测上清中IFN-α、IL-6、TNF-α的含量;实时定量聚合酶链反应(RT-PCR)检测pDCs的TLR9mRNA表达量。结果:①治疗前pDCs产生IFN-α、IL-6、TNF-α细胞因子的水平[(960.83±164.65)pg/ml,(156.15±39.89)pg/ml,(137.31±35.44)pg/ml]明显高于正常对照组[(616.67±105.98)pg/ml,(89.13±21.48)pg/ml,(88.53±25.81)pg/ml)](P<0.05);治疗后pDCs产生IFN-α、IL-6、TNF-α细胞因子水平分别降至(678.46±128.88)pg/ml、(97.77±26.31)pg/ml、(103.08±26.42)pg/ml,与治疗前比较差异有统计学意(P<0.05),与正常对照组比较,差异无统计学意义(P>0.05)。②治疗前pDCs的TLR9mRNA的表达水平高于正常对照组(P<0.05);治疗后pDCs的TLR9mRNA的表达水平低于治疗前(P<0.05),但与正常对照组的表达水平比较,差异无统计学意义(P>0.05)。结论:pDCs分泌的细胞因子及其表达的TLR9在ITP发病中起重要作用;地塞米松可能通过下调TLR9的表达,抑制pDCs分泌细胞因子的功能,从而对ITP起到治疗作用。
AIM: To investigate the effect of high dose dexamethasone on the function of plasma-like dendritic cells (pDCs) and the expression of Toll-like receptor 9 in patients with idiopathic thrombocytopenic purpura (ITP). Methods: Fifteen newly diagnosed ITP patients were given dexamethasone 40mg / d for 4 days. Peripheral blood plasma-like dendritic cells (pDCs) were isolated from 15 normal controls and 13 treated patients before and after treatment by immunomagnetic beads. Peripheral blood pDCs were stimulated with CPG-ODN 2216 and co-cultured for 24 h. The levels of IFN-α, IL-6 and TNF-α in supernatants were detected by enzyme-linked immunosorbent assay (ELISA). The expression of TLR9 mRNA in pDCs was detected by real-time quantitative polymerase chain reaction. Results: ① The levels of IFN-α, IL-6 and TNF-α produced by pDCs before treatment [(960.83 ± 164.65) pg / ml, (156.15 ± 39.89) pg / ml and (137.31 ± 35.44) pg / ml] Was significantly higher than that of the normal control group [(616.67 ± 105.98) pg / ml, (89.13 ± 21.48) pg / ml, (88.53 ± 25.81) pg / ml) (P <0.05), and the levels of TNF-α and cytokines decreased to (678.46 ± 128.88) pg / ml and (97.77 ± 26.31) pg / ml and (103.08 ± 26.42) pg / ml, respectively <0.05). Compared with the normal control group, the difference was not statistically significant (P> 0.05). ② The expression level of TLR9 mRNA in pDCs before treatment was higher than that in the normal control group (P <0.05). The expression of TLR9 mRNA in pDCs was lower than that before treatment (P <0.05), but the difference was not significant Statistical significance (P> 0.05). CONCLUSION: Cytokines secreted by pDCs and its expression of TLR9 play an important role in the pathogenesis of ITP. Dexamethasone may play a therapeutic role in ITP by down-regulating the expression of TLR9 and inhibiting the secretion of cytokines by pDCs.