1,25-(OH)_2D_3对激素抵抗型哮喘患者T淋巴细胞JNK/AP-1和糖皮质激素受体的影响  

Effect of 1,25-(OH)_2D_3 on JNK/AP-1 and glucocorticoid receptor of T lymphocytes in steroid-resistant asthmatics

作　　者：杨海华[1] 龙丰[1] 李圣青[2] 张霞[1] 金先桥[2] YANG Haihua LONG Feng LI Shengqing ZHANG Xia JIN Xianqiao(Department of Respiratory Medicine, Huashan Hospital North, Fudan University, Shanghai, 201907, P.R.China Department of Respiratory Medicine, Huashan Hospital, Fudan University, Shanghai, 200040, P.R,China)

机构地区：[1]复旦大学附属华山医院北院呼吸科,上海201907 [2]复旦大学附属华山医院呼吸科,上海200040

出　　处：《中国呼吸与危重监护杂志》2017年第2期155-159,共5页Chinese Journal of Respiratory and Critical Care Medicine

基　　金：上海市卫生和计划生育委员会青年科研项目(20134Y003)

摘　　要：目的观察激素抵抗型支气管哮喘(简称SR哮喘)患者外周血维生素D水平,探讨维生素D对SR哮喘患者T淋巴细胞JNK/AP-1和糖皮质激素受体的影响。方法收集2014年至2015年期间门诊和住院急性发作的哮喘患者62例,其中激素敏感型支气管哮喘(简称SS哮喘)26例,SR哮喘36例;选取同期健康体检正常者25例作为对照。收集入组者临床资料,取外周静脉血检测25-羟维生素D[25-(OH)D]水平并分离T淋巴细胞。T淋巴细胞培养分组如下:健康对照组(A组),SS哮喘对照组(B组),SR哮喘对照组(C组),SR哮喘JNK抑制剂(SP600125)+1,25-(OH)2D3组(D组),SR哮喘JNK抑制剂组(SP600125)(E组),以及SR哮喘1,25-(OH)2D3组(F组)。T淋巴细胞共培养48 h,培养结束后采用Western blot方法检测T淋巴细胞中磷酸化JNK(p-JNK)和磷酸化糖皮质激素受体(p-GR)的表达,RT-PCR法检测c-Jun m RNA的表达。结果 B组和C组血清25-(OH)D水平均低于A组(P<0.05),且C组低于B组(P<0.05)。B组和C组T淋巴细胞中p-JNK蛋白水平高于A组(P<0.05),且C组高于B组(P<0.05),E组和F组均低于C组(P<0.05),D组低于F组(P<0.05)。C组T淋巴细胞中p-GR蛋白低于A组和B组(P<0.05),E组和F组均高于C组(P<0.05),D组高于F组(P<0.05)。RT-PCR法检测B组和C组T淋巴细胞中c-Jun m RNA水平高于A组(P<0.05),C组高于B组(P<0.05),E组和F组均低于C组(P<0.05),D组低于F组(P<0.05)。C组患者25-(OH)D水平与p-JNK和c-Jun m RNA水平呈负相关(r=–0.69,r=–0.65,P<0.05),与p-GR水平呈正相关(r=0.72,P<0.05)。结论维生素D缺乏在SR哮喘患者中患病率高。1,25-(OH)2D3可通过抑制SR哮喘T淋巴细胞JNK/AP-1信号通路而促进p-GR的表达,这可能是维生素D改善SR哮喘患者的糖皮质激素抵抗的机制之一。Objective To observe the level of vitamin D in patients with steroid resistant （SR） asthma, and investigate the effect of 1,25-（OH）2D3 on JNK/AP-1 and glucocorticoid receptor of T lymphocytes in SR asthmatics. Methods Sixty-two outpatients and inpatients with asthma with acute exacerbation between 2014 and 2015 were recruited in the study, including 26 cases of steroid sensitive （SS） asthmatics and 36 cases of SR asthmatics. Meanwhile 25 healthy volunteers were recruited as control. Clinical data were collected and peripheral venous blood was sampled for measuring the level of 25-（OH）D and separating the T lymphocytes. T lymphocytes were assigned to six groups, ie. a healthy control group （Group A）, a SS asthmatics control group （Group B）, a SR asthmatics control group （Group C）, a SR asthmatics with JNK inhibitor （SP600125）＋1,25-（OH）2D3 group （Group D）, a SR asthmatics with JNK inhibitor （SP600125） group （Group E）, and a SR asthmatics with 1,25-（OH）2D3 group （Group F）, T lymphocytes were cultured for 48 hours. By the end of culture, the expression of phospho-INK （p-JNK） and phospho-glucocorticoid receptor （p-GR） of T lymphocytes were detected by Western blot method, and the expression of c-Jun mRNA was detected by RT-PCR method. Results The level of 25-（OH） D was lower in Grotlp B and Group C than Group A （P〈0.05）, and lower in Group C than Group B （P〈0.05）. The level ofp-JNK was higher in Group B and Group C than Group A （P〈0.05）, higher in Group C than Group B （P〈0.05）, lower in Group E and Group F than Group C （P〈0.05）, lower in Group D than Group F （P〈0.05）. The level ofp-GR was lower in Group C than Group A and Group B （P〈0.05）, higher in Group E and Group F than Group C （P〈0.05）, higher in Group D than Group F （P〈0.05）. The level ofc-Iun mRNA was higher in Group B and Group C than Group A （P〈0.05）, higher in Group C than Group B （P〈O.05）, lower in Group E and Group F than

关 键 词：维生素D 激素抵抗型支气管哮喘 T淋巴细胞 JNK AP-1 糖皮质激素受体 

分 类 号：R562.25[医药卫生—呼吸系统]