细胞因子信号抑制因子在实验性自身免疫性葡萄膜视网膜炎外周血单个核细胞内的表达变化

目的 观察和探讨细胞因子信号抑制因子（SOCS)在实验性自身免疫性葡萄膜视网膜炎（EAU）外周血单个核细胞（PBMC）内的表达及意义。方法 光感受器间维生素A 类结合蛋白(IRBP)免疫100只Lewis大鼠诱导EAU动物模型，随机分为对照组和治疗组。治疗组从免疫后第1天至第28天，给予环孢霉素A（CSA）灌胃，20 mg/（kg·d）；对照组给予等量生理盐水。免疫前、免疫后7、14、21、28 d采用裂隙灯显微镜观察大鼠眼部变化并抽取其心脏血，酶联免疫吸附试验（ELISA）方法检测血清白细胞介素（IL）-4、IL-12、干扰素（IFN）-γ的含量；实时荧光定量聚合酶链反应（PCR）和蛋白免疫印迹法检测PBMC内SOCS mRNA和蛋白表达。结果免疫后14 d 时炎症最明显。对照组可见明显的虹膜睫状体炎，治疗组前房轻微炎性渗出，但无虹膜后粘连及前房积脓。对照组血清中 IL-12、IFN-γ免疫后14 d达到最高峰，28 d下降到基础水平，治疗组在免疫后14 d达到高峰，但是升高程度明显低于对照组，其他时间点与免疫前比较无差异；IL-4在对照组中下降不明显，治疗组在整个病程中呈上升趋势。SOCS1、SOCS5 mRNA免疫后14 d表达最高，对照组分别是免疫前的4.05和3.83倍，治疗组分别为1.15和1.16倍；两组中含SH2结构的细胞因子诱导蛋白（CIS）与SOCS3 mRNA轻度升高，对照组较治疗组略明显。对照组中SOCS1、SOCS5蛋白在免疫后7、14、21 d时显著高于免疫前，CIS、SOCS3蛋白在免疫后14、21 d显著高于免疫前；治疗组中仅SOCS1蛋白在免疫后14 d显著高于免疫前，其他时间点较免疫前无明显改变。 结论 SOCS1、SOCS5水平升高与EAU发生过程中的Th1型反应增强有关，CIS、SOCS3水平轻度升高可能是存在着Th2细胞反应的增强以对抗Th1细胞反应，实现动态免疫平衡。

Expression of suppressor of cytokine signaling in peripheral blood mononuclear cells of experimental autoimmune uveoretinitis

Objective To observe the expression and investigate the significance of suppressor of eytokine signaling(SOCS)in peripheral blood mononuclear cells(PBMC)of experimental autoimmune uveitis(EAU). Methods 100 Lewis rats were immunized with interphotoreceptor retinoid-binding protein(IRBP)to induce EAU animal model,and they were divided into control group and treatment immunization;the control group received saline buffer at equal quantity.All eyes were evaluated by slitlamp microscopy before and after 7,14,21,28 days of immunization;IL-4,IL-12,IFN-γ in the serum were measured by enzyme linked immunosorbent assay(ELISA);the SOCS mRNA and protein level in PBMC were measured by quantitative polymerase chain reaction(q-PCR)and western blot. Results The inflammation was most obvious at 14 days after immunization.The controI group showed obvious iridocyclitis;the treatment group showed mild anterior chamber inflammation but no posterior synechia and hypopyon.The highest level of IL-12 and IFN-γ were observed at 14 days after immunization.followed by decline to the baseline at 28 days after immunization in control group; the highest level of IL12 and IFN-γ were found at 14 days after immunization in treatment group,but the ievel was lower than control group obviously.Compared with the level before immunization,there are no differences at other time-point.The concentration of IL-4 decreased indistinctly in control group but increased in treatment group.SOCS1、Both of SOCS1 and SOCS5 increased tO the highest level at 14 days after immunization,as 4.05 and 3.83 times of pre-immunization in control group respectively.as 1.15 and 1.16 times in treatment group respectively.The CIS and SOCS3 mRNA increased lightly in two groups and treatment group milder than control group.Marked increased expression of SOCSl and SOCS5 protein was detected at 7,1 4,2 1days than pre-immunization,both of CIS and SOCS3 protein were significantly increased on 14,21 days in control group;only SOCS1 protein was significantly increased on 14 days in treatment group and there are no differences at other time-point compared to pre-immunization. Conclusion UPregulation of SOCS1 and SOCS5 expression maybe related to intensive response of Th1 in the development of EAU.Mild up-regulation of CIS and SOCS3 maybe associated with intensive response of Th2 which against the reaction of Th1 to carry OUt the dynamic immune balance.