摘要
目的研究长链非编码RNA LINC01503在哮喘患儿中的表达及在巨噬细胞极化中的调控作用,为完善哮喘发病机制及发现新的防治靶点提供实验依据。方法纳入2017年8月26日至2018年1月11日复旦大学附属儿科医院(我院)呼吸科门诊确诊的哮喘患儿(哮喘组)及我院健康体检儿童(健康对照组),分离2组外周血中的单个核细胞,采用RTq PCR检测LINC01503在两组中的表达; LPS与IL-4分别诱导巨噬细胞向M1及M2方向极化,采用RT-qPCR检测LINC01503的动态变化;脂质体转染siRNA敲低LINC01503表达,采用RT-qPCR及Western blot检测LINC01503对巨噬细胞极化的影响。结果哮喘组28例,健康对照组46例。与健康对照组相比,LINC01503在哮喘组中的表达显著升高。LINC01503在M1巨噬细胞中明显下调,而在M2巨噬细胞中明显上调。siRNA敲低实验发现,LINC01503促进M1巨噬细胞的极化,上调M1标志基因如IL-6、TNF-α和CXCL10等的表达; LINC01503抑制M2巨噬细胞的极化,下调M2标志基因如CD206和CD209的表达。Western blot发现LINC01503主要通过促进细胞外调节蛋白激酶(ERK)的磷酸化而影响巨噬细胞极化。结论 LINC01503在哮喘患儿中高表达,通过负反馈调控巨噬细胞的活化,有望成为哮喘治疗的新靶点。
Objective To explore the potential role of LINC01503 in the regulation of macrophage polarization and its function in the pathology of children asthma to achieve a better understanding of the pathology of asthma and find new strategies for asthma treatment.Methods Blood was collected from asthmatic patients and healthy controls from August 26,2017 to January 11,2018 at Children's Hospital of Fudan University.PBMCs were separated from blood by gradient centrifuge.RT-qPCR was used for the detection of the expression of LINC01503 in PBMCs.LPS and IL-4 were used for the inducing of M1 and M2 polarization of macrophage separately.siRNA was used for knocking down LINC01503.RT-qPCR and Western blot were used for the detection of the effect of LINC01503 on the regulation of macrophage polarization.Results We enrolled 28 patients of asthma and 46 healthy controls.Comparing with healthy controls,the expression of LINC01503 was significantly enhanced in asthmatic patients.LINC01503 was up-regulated in M2 macrophage but down-regulated in M1 macrophage.LINC01503 inhibited the M2 macrophage polarization indicated as the down regulation of CD206 and CD209.On the opposite,LINC01503 promoted the M1 macrophage polarization indicated as the up regulation of the expression of inflammatory mediators such as IL-6,TNF-α and CXCL10 through enhancing the phosphorylation of ERK.Conclusion LINC01503 was up-regulated in the asthmatic patients and it regulated macrophage polarization through a negative feedback loop.
作者
夏莉
刘丽娟
王香
孙立成
付劲蓉
韩晓
钱莉玲
周玉峰
XIA Li;LIU Li-juan;SUN Li-cheng;FU Jin-rong;HAN Xiao;QIAN Li-ling;ZHOU Yu-feng(Children's Hospital and Institutes of Biomedical Sciences of Fudan University,Shanghai 201102,China;Key Laboratory of Neonatal Diseases,Ministry of Health,Shanghai 201102,China)
出处
《中国循证儿科杂志》
CSCD
北大核心
2019年第1期49-53,共5页
Chinese Journal of Evidence Based Pediatrics
基金
国家自然科学基金面上项目:81671561
科技部国家重点研发计划:2016YFC1305102
上海市卫生和计划生育委员会科研课题:201740065
浦江人才计划项目:16PJ1401600
国家儿童医学中心"海聚"国际联合实验室项目:EK1125180109
