miR-148a-3p对银屑病患者Th细胞活化调控的影响  

Effects of miR-148a-3p on the Regulation of Th Cell Activation in Patients with Psoriasis

作　　者：林玲[1] 曾菁莘 张锡宝[1] 罗权 LIN Ling;ZENG Jingxin;ZHANG Xinbao;LUO Quan(Department of Dermatology,Guangzhou Institute of Dermatology,Guangzhou 510095,China;Department of Dermatology,Guangdong No.2 Provincial People's Hospital,Affiliated to Southern Medical University,Guangzhou 510317,China)

机构地区：[1]广州市皮肤病防治所,广东广州510095 [2]广东省第二人民医院,广东广州510317

出　　处：《中国皮肤性病学杂志》2020年第7期743-747,共5页The Chinese Journal of Dermatovenereology

基　　金：广东省医学科研基金(A2016542);广东省企业技术研发与省级改造专项资金项目(2013B021800044);广东省医学科研基金(A2015305)。

摘　　要：目的 在前期研究的基础上进一步验证miR-148a-3p通过促凋亡蛋白Bim对银屑病发病机制中T细胞亚群分化的影响及相关免疫应答机制的调控。方法 培养人T淋巴白血病(Jurkat)细胞;Bim siRNA、miR-148a-3p mimics、miR-148a-3p inhibitor,分别电转CD4+T细胞,RT-qPCR检测miR-148a-3p和Bim mRNA,Western blot检测 Bim 蛋白;通过ELISA检测培养上清中细胞因子含量。结果 Bim siRNA、miR-148a-3p mimics、miR-148a-3p inhibitor分别电转CD4+T细胞(Jurkat细胞),电转效率可达84.5%。与空白组相比,过表达的miR-148a-3p可有效抑制靶基因Bim mRNA及其蛋白的表达,ELISA检测培养上清中IFN-γ及IL-17均明显升高,IL-4及IL-10均明显降低(P均<0.05);而抑制miR-148a-3p时Bim mRNA及其蛋白的表达升高,IFN-γ明显减少(P均<0.05),而IL-17表达(P>0.05)下降不明显,IL-4及IL-10均明显升高(P均<0.05)。在正常CD4+T细胞中沉默Bim的表达,IFN-γ及IL-17亦显著升高(P均<0.05)。结论 在银屑病患者中高表达的miR-148a-3p靶向调控Bim的表达可导致Th1、Th17细胞异常活化,Th2细胞数量减少及Treg细胞免疫抑制功能减弱,这提示miR-148a-3p参与寻常性银屑病病情的发生发展,为后续miR-148a-3p靶向干预寻常性银屑病病情提供了依据。Objective On the basis of previous studies,the effect of miR-148a-3p on the differentiation of T cell subsets in the pathogenesis of psoriasis and the regulation of related immune response were further verified by the proapoptotic protein Bim.Methods Cultured human T lymphoblastic leukemia(Jurkat)cells;Bim siRNA,miR-148a-3p mimics,miR-148a-3p inhibitor,respectively,electroporated CD4+ T cells,RT-qPCR detection of miR-148a-3p and Bim mRNA,Western blot detection Bim protein;cytokine content in culture supernatant was determined by ELISA.Results Bim siRNA,miR-148a-3p mimics and miR-148a-3p inhibitor electroporated CD4+ T cells(Jurkat cells),respectively,and the electrotransfer efficiency was 84.5%.Compared with the blank group,over-expressed miR-148a-3p could effectively inhibit the expression of the target gene Bim mRNA and its protein.IFN-γ and IL-17 were significantly increased in the culture supernatant,while IL-4 and IL-10 were significantly decreased(P<0.05). The expression of Bim mRNA and its protein was increased when miR-148a-3p was inhibited,IFN-γ was significantly decreased(P<0.05),IL-17 was expressed(P>0.05),and IL-4 and IL-10 were significantly increased(P<0.05).The expression of Bim was silenced in normal CD4+ T cells,and IFN-γ and IL-17 were also significantly increased(P<0.05).Conclusion The high expression of miR-148a-3p in patients with psoriasis can regulate the expression of Bim,which leads to the abnormal activation of Th1 and Th17 cells,the decrease of Th2 cell number and the weakening of Treg cell immunosuppression,suggesting that miR-148a-3p is involved in vulgaris.The results provide a basis for subsequent miR-148a-3p targeted intervention for psoriasis vulgaris.

关 键 词：寻常性银屑病 miR-148a-3p CD4+T细胞 BIM蛋白 

分 类 号：R758.63[医药卫生—皮肤病学与性病学]