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目的阐明Th17细胞特异性RORγt、相关细胞因子IL-17A、IL-6和Treg细胞特异性FOXP3、相关细胞因子IL-10、TGF-β在川崎病不同时期的表达以及与冠状动脉损伤(CAL)、静脉注射免疫球蛋白(IVIG)治疗反应性的关系。方法选择85例川崎病患儿为病例组,30例健康儿童为对照组,病例组患儿于开始IVIG治疗前、治疗后3、21 d取外周血,采用酶联免疫吸附法(ELISA)检测外周血细胞因子IL-10、IL-17A、TGF-β及IL-6水平,实时PCR检测RORγt mRNA和FOXP3mRNA表达水平。结果病例组患儿治疗前血中IL-17A、IL-6比对照组儿童高,差异有统计学意义(P<0.01);IVIG治疗3、21 d后IL-6显著低于治疗前,差异有统计学意义(P<0.01)。血IL-17A在治疗3 d后有所下降,但与治疗前相比差异无统计学意义(P>0.05);治疗21 d后IL-17A显著下降,但仍高于对照组,差异有统计学意义(P<0.01)。IL-10明显高于对照组(P<0.01),而TGF-β与对照组差异无统计学意义(P>0.05);IVIG治疗3 d后IL-10无显著下降(P>0.05),TGF-β水平在IVIG治疗前、治疗后3、21 d差异无统计学意义(P>0.05)。与对照组儿童比,病例组患儿治疗前外周血细胞FOXP3 mRNA表达水平差异无统计学意义(P>0.05),治疗3 d时FOXP3 mRNA表达水平显著上升,RORγt mRNA显著升高,差异均有统计学意义(P<0.01);治疗3、21 d时Th17的RORγt mRNA表达水平均有所下降,仍高于对照组儿童。川崎病CAL患儿IL-10、IL-17A均高于无CAL患儿,IVIG无反应型的川崎病患儿IL-10及IL-17A均高于IVIG敏感型的患儿,差异均有统计学意义(P<0.01)。结论川崎病患儿急性期外周血中Th17的转录因子RORγt以及细胞因子IL-17A、IL-6升高,而Treg细胞特异性FOXP3及其细胞因子IL-10降低;川崎病患儿外周血IL-17A的持续高水平与川崎病IVIG无反应以及CAL有关。本研究为阐明川崎病的发病机制以及治疗提供了科学依据,为IVIG无反应以及IVIG治疗的敏感性判断提供了潜在的标志物。
Objective To investigate the expression of Th17-specific RORγt, related cytokines IL-17A, IL-6 and Treg cell-specific FOXP3, related cytokines IL-10 and TGF-β in patients with Kawasaki disease at different stages and with coronary artery injury (CAL) , Intravenous immunoglobulin (IVIG) treatment response relationship. Methods 85 children with Kawasaki disease were selected as case group and 30 healthy children as control group. Peripheral blood was taken at 3 and 21 days after IVIG treatment and before treatment with IVIG in the case group. Enzyme-linked immunosorbent assay (ELISA) The levels of IL-10, IL-17A, TGF-β and IL-6 in peripheral blood were detected by real-time PCR. The expression of RORγt mRNA and FOXP3 mRNA were detected by real-time PCR. Results The blood levels of IL-17A and IL-6 in children before treatment were significantly higher than those in the control group (P <0.01). IL-6 in IVIG treatment group was significantly lower than that before treatment There was statistical significance (P <0.01). Blood IL-17A decreased after 3 days of treatment, but there was no significant difference between before and after treatment (P> 0.05); IL-17A decreased significantly after 21 days of treatment, but still higher than that of control group Significance (P <0.01). IL-10 was significantly higher than that of the control group (P <0.01), but there was no significant difference between the TGF-β and the control group (P> 0.05) There was no significant difference in the levels of -β before and after IVIG 3 and 21 days (P> 0.05). Compared with the control group, there was no significant difference in the expression of FOXP3 mRNA in the peripheral blood cells before treatment in the case group (P> 0.05). The FOXP3 mRNA level and the RORγt mRNA level were significantly increased at 3 d (P <0.01). At 3 and 21 d, the expression of RORγt mRNA in Th17 cells was still lower than that in the control group. IL-10 and IL-17A in Kawasaki disease CAL children were higher than those in children without CAL, IL-10 and IL-17A in children with IVIG non-responsive Kawasaki disease were higher than those in IVIG-sensitive children Significance (P <0.01). CONCLUSIONS: Th17 transcription factor RORγt and cytokines IL-17A and IL-6 are increased in peripheral blood of children with Kawasaki disease in the acute phase, while Treg cell-specific FOXP3 and its cytokine IL-10 are decreased. Peripheral blood IL Sustained high levels of -17A were unrelated to Kawasaki disease IVIG and CAL. This study provides a scientific basis for clarifying the pathogenesis and treatment of Kawasaki disease and provides a potential marker for the non-response of IVIG and the sensitivity of IVIG treatment.