儿童白癜风外周血单个核细胞中CXC趋化因子10、趋化因子受体3及干扰素-γ表达研究

Expression of chemokine 10,chemokine receptor 3 and interferon-γ in the peripheral blood mononuclear cells of children with vitiligo

目的:检测寻常型白癜风患儿外周血中CXC趋化因子(CXCL)10、趋化因子受体(CXCR)3及干扰素(IFN)-γ含量及其mRNA和蛋白表达水平。方法:选取寻常型白癜风患儿100例(其中进展期及稳定期各50例)及正常对照组50例。采用酶联免疫吸附试验(ELISA)检测2组CXCR3、CXCL10及IFN-γ含量;提取外周血单个核细胞(PBMC),各组随机抽取20例采用反转录聚合酶链式反应(RT-PCR)及western blot分别检测CXCR3、CXCL10及IFN-γmRNA和蛋白表达水平;将各组余30例PBMC进行体外培养,设2个复孔(一个孔为空白,另一孔加入抗IFN-γ抗体干预),作用24 h后,采用ELISA法检测培养上清液中CXCR3、CXCL10和IFN-γ含量。结果:与正常对照组比较,2组患儿外周血及外周血PBMC上清液中CXCR3、CXCL10及IFN-γ含量均显著升高(P<0.05),且PBMC中CXCR3、CXCL10及IFN-γmRNA和蛋白表达水平均显著升高(P<0.05)。与稳定期组比较,进展期组患儿外周血及PBMC上清液中CXCR3、CXCL10和IFN-γ含量均显著升高(P<0.05),且外周血PBMC中CXCR3、CXCL10及IFN-γmRNA和蛋白表达水平均显著升高(P<0.05)。加入抗IFN-γ抗体干预后,各组PBMC上清液中CXCR3、CXCL10及IFN-γ含量均显著降低(P<0.05)。结论:儿童白癜风的发生与外周血高表达CXCR3、CXCL10及IFN-γ相关,IFN-γ的高表达可能是上调CXCR3和CXCL10表达的调控因素之一,具体作用机制有待进一步研究。

Objective:To study the expression of chemokine 10(CXCR10),chemokine receptor 3(CXCR3)and interferon-γ(IFN-γ)in the peripheral blood of children with vitiligo vulgaris.Methods:One hundred patients with vitiligo vulgaris were selected,of which 50 cases were as progressive group,and the other 50 cases were as stable group.Fifty children without vitiligo were selected as normal control group.ELISA was used to assess the levels of CXCR3,CXCL10 and IFN-γexpression in all groups.Peripheral blood mononuclear cells(PBMCs)were extracted,and 20 cases were randomly selected from each group to evaluate the mRNA and protein expression of CXCR3,CXCL10 and IFN-γby RT-PCR and western blotting,respectively.The PBMCs of remaining 30 cases were cultured in vitro,and two holes were established(one hole with no treatment,the other one with IFN-γtreatment).After 24 hours,the levels of CXCR3,CXCL10 and IFN-γin culture supernatant were measured by ELISA.Results:Compared with the normal control group,the expression levels of CXCR3,CXCL10 and IFN-γin peripheral blood and culture supernatant of PBMCs in vitiligo patients,both progressive group and stable group,were significantly increased(P<0.05).The mRNA and protein expression levels of CXCR3,CXCL10 and IFN-γin PBMCs of vitiligo patients,both progressive group and stable group,were also significantly increased(P<0.05),compared with that of control group.Compared with the stable group,the levels of CXCR3,CXCL10 and IFN-γexpression in peripheral blood and culture supernatant of PBMCs in progressive group were significantly increased(P<0.05),and the levels of mRNA and protein expression of CXCR3,CXCL10 and IFN-γin PBMCs were significantly increased(P<0.05).After anti-IFN-γintervention,the levels of CXCR3,CXCL10 and IFN-γin PBMCs culture supernatant of vitiligo patients,both groups,were significantly decreased(P<0.05).Conclusions:The occurrence of vitiligo in children is associated with high expression of CXCR3,CXCL10 and IFN-γin peripheral blood.The high expression of IFN-γmay be one of the regulatory factors that up-regulate the expression of CXCR3 and CXCL10.Further studies are necessary to determine the mechanism of action.