IL-35通过调控STAT1信号通路在儿童哮喘中的作用研究

The role of IL-35 in regulation of STAT1 signaling pathway in childhood asthma

目的探究白细胞介素35(IL-35)在哮喘患儿血清和单个核细胞中的表达量以及作用机制。方法收集38例哮喘儿童为研究对象,并选取40例健康儿童为对照组。酶联免疫吸附测定法(ELISA)测定血清中白细胞介素4(IL-4)、免疫球蛋白E(Ig E)、IL-35的水平,实时荧光定量PCR(qRT-PCR)检测单个核细胞中IL-35 mRNA和信号转导和转录激活因子1(STAT1)mRNA的表达,蛋白质免疫印迹法(Western blot)检测单个核细胞中STAT1蛋白的表达量,并分析哮喘患儿中IL-35与IL-4、Ig E、STAT1的相关性。结果与健康对照组相比,哮喘患儿血清中IL-4、Ig E的水平显著增加(P<0.05),二者呈显著的正相关性;哮喘组单个核细胞中IL-35 mRNA和血清中IL-35蛋白水平均显著低于健康对照组(P<0.05),血清中IL-35与IL-4呈显著的负相关,与Ig E无显著相关性;STAT1 mRNA和STAT1蛋白在哮喘患者单个核细胞中的表达量显著高于健康对照组(P<0.05),IL-35 mRNA与STAT1 mRNA呈显著负相关。结论 IL-35在哮喘患者的表达量显著下调,可能通过阻碍IL-4的表达,抑制STAT1信号通路的激活,从而阻碍哮喘病的发生、发展。

Objective To investigate the expression and mechanism of IL-35 in childhood asthma.Methods The levels of IL-4,Ig E and IL-35 in serum were measured by enzyme-linked immunosorbent assay(ELISA) of 38 patients with asthma(asthmatic children group) and40 normal subjects(healthy control group).Quantitative real-time polymerase chain reaction(qRT-PCR) were used to detect the expression of IL-35 mRNA and STAT1 mRNA in peripheral biood mononuclear cells.The expression of STAT1 protein in peripheral biood mononuclear cells were detected by Western blot.The correlation between the IL-35 and IL-4,Ig E and STAT1 in childhood asthma were analyzed.Results Compared with the healthy control group,the levels of IL-4 and Ig E in serum of asthmatic children were significantly increased(P < 0.05),and IL-4 was positively correlated with Ig E.The levels of IL-35 mRNA in peripheral biood mononuclear cells and IL-35 protein in serum of asthmatic children were lower than those in healthy controls(P < 0.05),IL-35 were negatively correlated with IL-4 in serum.The expression of STAT1 mRNA and STAT1 protein in peripheral biood mononuclear cells of asthmatic patients were significantly higher than those in healthy controls(P < 0.05).IL-35 mRNA was negatively correlated with STAT1 mRNA in asthmatic children.Conclusion The expression of IL-35 decreases in asthmatic children.The general steps of the production of asthma can be controlled by inhibiting the STAT1 activity,and down regulating the expressions of IL-4,so childhood asthma can be treated more effectively by this mechanism.