GPC3-WNT-JNK通路介导脂多糖诱导肺局部细胞炎症  

Glypican3-WNT-JNK pathway mediates the molecular mechanism of local cell inflammation in lung induced by lipopolysaccharide

作　　者：陈超蕾[1] 林雨婷[1] 张桑桑 宋晨剑 何飞 董莉[1] 陈俊杰[1] 王蓓蓓 董年 李玉苹[1] 陈成水[1] CHEN Chaolei;lin Yuting;ZHANG Sangsang;SONG Chenjian;HE Fei;DONG Li;CHEN Junjie;WANG Beibei;DONG Nian;LI Yuping;CHEN Chengshui(Department of Respiratory and Critical Care Medicine,the First Affiliated Hospital of Wenzhou Medical University,Wenzhou325015,China;Department of Cardiovascular Medicine,Wenzhou Hospital of Integrated Traditional Chinese and Western Medicine,Wenzhou 325000,China)

机构地区：[1]温州医科大学附属第一医院呼吸与危重症医学科,浙江温州325015 [2]温州市中西医结合医院心血管内科,浙江温州325000

出　　处：《温州医科大学学报》2019年第10期703-711,共9页Journal of Wenzhou Medical University

基　　金：国家自然科学基金资助项目(81600062,81770074);温州市公益性科技计划项目(Y20150129,Y20180125);浙江省医药卫生科技计划项目(2018264229);浙江省自然科学基金资助项目(LY18H010006);温州医科大学附属第一医院科研孵化项目(FHY2015037)

摘　　要：目的:探讨GPC3在脂多糖(LPS)诱导的气道上皮细胞局部炎症微环境中的重要作用,并以GPC3-WNT通路介导的调控机制为核心进一步研究LPS诱导的肺局部炎症分子机制。方法:LPS经气道滴入小鼠后模拟肺损伤模型,检测肺泡灌洗液及肺组织中GPC3表达情况以及其表达位置。培养人肺16HBE细胞,LPS刺激16HBE细胞建立急性炎症细胞,予以RT-PCR检测其细胞中GPC3、TNF-α和TGF-β的mRNA表达情况,予以WesternBlot检测GPC3、TGF-β和TNF-α蛋白表达情况。加入外源性GPC3刺激16HBE细胞,建立GPC3高表达细胞模型,检测炎症因子TNF-α和TGF-β mRNA的表达情况。不同浓度GPC3-WNT通路抑制剂处理细胞,观察上述指标变化。结果:LPS诱导ALI小鼠模型显示GPC3表达水平显著提高。此外,LPS在体外也产生了GPC3表达升高的现象。同时,外源性GPC3蛋白刺激支气管上皮细胞产生多种炎症因子,可能提示GPC3具有促炎作用。此外,GPC3诱导支气管上皮细胞系炎性细胞因子的产生被WNT-JNK通路的抑制剂阻断,提示ALI/ARDS过程中GPC3参与肺局部的潜在信号通路。结论:LPS刺激支气管上皮细胞的过程中存在一条线性信号通路即GPC3-WNT-JNK。这一发现可能为ALI/ARDS的治疗和生物标志物的开发提供一个新的分子靶点和分子机制。Objective: To investigate the important role of(glypican-3) GPC3 in lipopolysaccharide(LPS)-induced microenvironment of local inflammation in airway epithelial cells, and to further investigate the GPC3-WNT pathway molecular mechanism of LPS-induced local inflammation in the lung. Methods: The expression of GPC3 in alveolar lavage and lung tissue and its expression location were detected in an ALI mouse model simulated by LPS airway aspiration. Cultured human lung 16 HBE cells were stimulated by LPS to establish acute inflammatory cells. The mRNA expressions of GPC3, TNF-α and TGF-β were detected by RT-PCR, meanwhile the expressions of GPC3, TNF-α and TGF-β proteins were assessed by Western Blot. The expression of inflammatory cytokines TNF-α and TGF-β was detected in 16 HBE cells stimulated by exogenous GPC3. Cells were treated with GPC3-WNT pathway inhibitor at different concentrations, then expression levels of TNF-α and TGF-β were measured by the above-mentioned methods. Results: GPC3 expression showed a significant increase in ALI mouse model induced by LPS. In addition, increased expression of GPC3 was also observed in LPS in vitro. Meanwhile, exogenous GPC3 protein stimulated bronchial epithelial cells to produce a variety of inflammatory factors, which may indicate that GPC3 has pro-inflammatory effect. Additionally, the production of inflammatory cytokines in bronchial epithelial cell induced by GPC3 was blocked by inhibitors of the WNT-JNK pathway, suggesting that GPC3 involved in the potential signaling pathway in local lung cells during ALI/ARDS. Conclusion: There is a linear signal pathway, GPC3-WNT-JNK, in the process of LPS stimulating bronchial epithelial cells. This discovery may provide a new molecular target and molecular mechanism for the treatment of ALI/ARDS.

关 键 词：急性肺损伤 急性呼吸窘迫综合征 支气管上皮细胞 GLYPICAN-3 WNT JNK 炎症 

分 类 号：R563.1[医药卫生—呼吸系统]