细胞因子信号抑制因子在Vogt-小柳原田病患者外周血单个核细胞的表达

Expression of suppressor of cytokine signaling in peripheral blood monocular cells of patients with Vogt-Koyanagi-Harada disease

目的探讨Vogt-小柳原田病（Vogt-Koyanagi-Harada，VKH）患者外周血单个核细胞（peripheral blood mononuclear cell，PBMC）细胞因子信号抑制因子（suppressor of cytokine signaling，SOCS）的表达。方法本研究设计为病例对照研究。抽取15例VKH活动期患者、17例静止期患者和16例正常人的外周血，用酶联免疫吸附测定方法检测血清中细胞因子IL-4、IL-12、IFN-γ的水平；用实时荧光定量PCR和蛋白免疫印迹方法检测外周血单个核细胞内SOCS基因和蛋白的表达，对三组实验结果数据采用方差分析中的两两比较进行统计。结果活动期和静止期VKH患者及正常对照组血清中IL-4水平分别为（28．40±5．93）、（34．5±9．47）和（11．25±4．43）ng／L，IL-12的水平为（24．33±8．55）、（11．53±6．11）和（5．19±2．43）ng／L，IFN-γ的水平为（18．05±2．23）、（15．53±2．63）和（1．61±3．47）ng/L，IL-4、IL-12和IFN-γ在活动期和静止期VKH患者血清中的水平均较正常对照组显著升高（P〈0．01）。IL4在静止期患者较活动期患者水平升高（P〈0．01）。IL-12和IFN-γ在静止期患者较活动期患者水平低（P〈0．01，P〈0．05）。SOSC基因表达的检测结果显示含SH2结构的细胞因子诱导蛋白（cytokine inducible SH2 containing protein，CIS）、SOCS1、SOCS2、SOCS3、SOCS5mRNA在VKH活动期患者PBMC内的表达分别是正常对照组的0．72、4．92、1．09、0．75和1．15倍，静止期患者是正常对照组的1．15、2．25、1．40、0．69和1．16倍。蛋白免疫印迹结果显示活动期患者的CIS水平显著低于静止期患者和正常人（P〈0．01），静止期和正常对照组之间差异无统计学意义（P〉0．05）；SOCS1在活动期患者体内显著高于静止期患者和正常对照组（P〈0．01，P〈0．05），静止期和正常对照组之间差异无统计学意义（P〉0．05）；SOCS2在患者和正常人之间无明显差别（均P〉0．05）；活动期和静止期患者的SOCS3均明显较正常对照组降低（P〈0．05）；SOCS5在两组患者体内的水平均高于正常对照组（P〈0．01），且静止期患者的水平显著高于活动期（P〈0．05）。结论VKH患者PBMC内SOCS1和SOCS5水平升高、CIS和SOCS3水平降低与VKH发生过程中的Th1型反应增强有关。SOCS5在葡萄膜炎发生过程中可能起保护作用。临床上静止期的葡萄膜炎患者体内的免疫环境仍处于异常状态，这可能是葡萄膜炎易复发和慢性化的原因。

Objective To investigate suppressor of cytokine signaling （SOCS） mRNA and protein expression in peripheral blood mononuclear cells （PBMC） of patients with Vogt-Koyanagi-Harada （VKH） disease. Methods Blood samples were taken from 15 VKH patients with active uveitis, 17 quiescent patients and 16 healthy individuals. IFN-7, IL-12 and IL-4 in the serum were measured by ELISA. PBMC were subjected to analysis of SOCS mRNA and protein expression using quantitative RT-PCR and western blot, respectively. Results The level of IL-γ in the serum of VKH patients and in controls were （28.40 ± 5.93 ） ng/L, （34. 5 ±9.47） ng/L and （ 11.25± 4.43） ng/L, IL-12 were （24. 33 ±8.55 ） ng/L, （ 11.53 ±6.11）ng/L and （5.19 ± 2. 43 ） ng/L, IFN-γ were （18.05±2.23）ng/L, （15.53±2.63）ng/L and （1.61± 3.47）ng/L, respectively. The level of IFN-γ, IL-12 and IL-4 were all significantly higher in the serum of VKH patients than in controls （ P 〈 0.01 ）. IL-4 in quiescent patients was higher than in active patients （P 〈0. 01 ）, IL-12 and IFN-γ were lower in quiescent patients was higher than in active patients （P 〈 0. 01, P 〈 0. 05）. Cytokine inducible SH2 containing protein （CIS） mRNA, SOCS1 mRNA, SOCS2 mRNA, SOCS3 mRNA and SOCS5 mRNA levels in PBMC of VKH patients with active uveitis are 0. 72, 4.92, 1.09, 0.75 and 1.15 folds than that in healthy volunteers, respectively. They are 1.15, 2.25, 1.40, 0. 69 and 1.16 folds in static patients, respectively. Marked decreased expression of CIS protein is detected in both active and quiescent patients with no significant difference between two groups （ both P 〈 0. O1 ）. SOCS1 protein is up-regulated significantly in active patients compares to in quiescent patients nor in healthy volunteers （P 〈 0. 01, P 〈 0. 05 ）. SOCS3 protein is significantly decreased in patients than in controls（both P 〈0. 05）. SOCS5 protein is much higher in patients than in controls（both P 〈0.01 ）, and even higher in quiescent patients than in active episode （ P 〈 0. 05 ）. Conclusions Up regulation of SOCS1 and SOCS5 expression and down-regulation of SOCS3 and CIS may correlate with the development of a Thl mediated immune response in VKH disease. There is insidious inflammation in VKH patients with clinically quiescent uveitis, and this may be one of the causes of persistence and recurrences of uveitis.