GPC3-WNT-JNK通路介导脂多糖诱导肺局部细胞炎症

摘要
图/表
参考文献
相关文章 (15)

全文: PDF (2032 KB) HTML (1 KB)
输出: BibTeX | EndNote (RIS)

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Western Blot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。

	服务
	把本文推荐给朋友
	加入我的书架
	加入引用管理器
	E-mail Alert
	RSS
	作者相关文章
	陈超蕾
	林雨婷
	张桑桑
	宋晨剑
	何飞
	董莉
	陈俊杰
	王蓓蓓
	董年
	李玉苹
	陈成水

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

Abstract：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 16HBE 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 16HBE 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.

Key words： acute lung injury acute respiratory distress syndrome bronchial epithelial cells glypican-3 WNT JNK inflammation

收稿日期: 2019-01-11

基金资助: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）。

通讯作者: 陈成水，主任医师，Email：wzchencs@163.com。

作者简介: 陈超蕾（1987-），女，浙江台州人，住院医师，硕士。

链接本文:

https://xb.wmu.edu.cn/CN/ 或 https://xb.wmu.edu.cn/CN/Y2019/V49/I10/703

[1] ASHBAUGH D G, BIGELOW D B, PETTY T L, et al. Acute respiratory distress in adults[J]. Lancet, 1967, 2(7511): 319-323.
[2] BHATIA M, ZEMANS R L, JEYASEELAN S. Role of chemokines in the pathogenesis of acute lung injury[J]. Am J Respir Cell Mol Biol, 2012, 46(5): 566-572.
[3] JOHNSON E R, MATTHAY M A. Acute lung injury: epidemiology, pathogenesis, and treatment[J]. J Aerosol Med Pulm Drug Deliv, 2010, 23(4): 243-252.
[4] CHEN C, FANG X, WANG Y, et al. Preventive and therapeutic effects of phosphoinositide 3-kinase inhibitors on acute lung injury[J]. Chest, 2011, 140(2): 391-400.
[5] CHEN C, SHI L, LI Y, et al. Disease-specific dynamic biomarkers selected by integrating inflammatory mediators with clinical informatics in ARDS patients with severe pneumo-nia[J]. Cell Biol Toxicol, 2016, 32(3): 169-184.
[6] PILIA G, HUGHES-BENZIE R M, MACKENZIE A, et al. Mutations in GPC3, a glypican gene, cause the Simpson-Golabi-Behmel overgrowth syndrome[J]. Nat Genet, 1996, 12(3): 241-247.
[7] VEUGELERS M, VERMEESCH J, WATANABE K, et al. GPC4, the gene for human K-glypican, flanks GPC3 on xq26: deletion of the GPC3-GPC4 gene cluster in one family with Simpson-Golabi-Behmel syndrome[J]. Genomics, 1998, 53(1): 1-11.
[8] LIBBRECHT L, SEVERI T, CASSIMAN D, et al. Glypican-3 expression distinguishes small hepatocellular carcinomas from cirrhosis, dysplastic nodules, and focal nodular hyperplasia-like nodules[J]. Am J Surg Pathol, 2006, 30(11): 1405-1411.
[9] NASSAR A, COHEN C, SIDDIQUI M T. Utility of glypican-3 and survivin in differentiating hepatocellular carcinoma from benign and preneoplastic hepatic lesions and metastatic carcinomas in liver fine-needle aspiration biopsies[J]. Diagn Cytopathol, 2009, 37(9): 629-635.
[10] AVIEL-RONEN S, LAU S K, PINTILIE M, et al. Glypican-3 is overexpressed in lung squamous cell carcinoma, but not in adenocarcinoma[J]. Mod Pathol, 2008, 21(7): 817-825.
[11] LAPUNZINA P. Risk of tumorigenesis in overgrowth syndromes: a comprehensive review[J]. Am J Med Genet C Semin Med Genet, 2005, 137C(1): 53-71.
[12] BOSWELL S, SHARIF S, ALISA A, et al. Induction of latency-associated peptide (transforming growth factor-beta(1)) expression on CD4+T cells reduces Toll-like receptor 4 ligand-induced tumour necrosis factor-alpha production in a transforming growth factor-beta-dependent manner[J]. Immunology, 2011, 133(3): 278-287.
[13] CAPURRO M I, XIANG Y Y, LOBE C, et al. Glypican-3 promotes the growth of hepatocellular carcinoma by stimulating canonical Wnt signaling[J]. Cancer Res, 2005, 65(14): 6245-6254.
[14] STIGLIANO I, PURICELLI L, FILMUS J, et al. Glypican-3 regulates migration, adhesion and actin cytoskeleton organization in mammary tumor cells through Wnt signaling mod-ulation[J]. Breast Cancer Res Treat, 2009, 114(2): 251-262.
[15] CAPURRO M, MARTIN T, SHI W, et al. Glypican-3 binds to Frizzled and plays a direct role in the stimulation of canonical Wnt signaling[J]. J Cell Sci, 2014, 127(Pt 7): 1565-1575.
[16] MAGISTRI P, LEONARD S Y, TANG C M, et al. The glypican 3 hepatocellular carcinoma marker regulates human hepatic stellate cells via Hedgehog signaling[J]. J Surg Res, 2014, 187(2): 377-385.
[17] CAPURRO M, SHI W, IZUMIKAWA T, et al. Processing by convertases is required for glypican-3-induced inhibition of Hedgehog signaling[J]. J Biol Chem, 2015, 290(12): 7576-7585.
[18] KITTAKA N, TAKEMASA I, TAKEDA Y, et al. Molecular mapping of human hepatocellular carcinoma provides deeper biological insight from genomic data[J]. Eur J Cancer, 2008, 44(6): 885-897.
[19] LIU A, CHEN S, CAI S, et al. Wnt5a through noncanonical Wnt/JNK or Wnt/PKC signaling contributes to the differentiation of mesenchymal stem cells into type II alveolar epithelial cells in vitro[J]. PLoS One, 2014, 9(3): e90229.
[20] SABAPATHY K. Role of the JNK pathway in human dis-eases[J]. Prog Mol Biol Transl Sci, 2012, 106: 145-169.
[21] MINDEN A, KARIN M. Regulation and function of the JNK subgroup of MAP kinases[J]. Biochim Biophys Acta, 1997, 1333(2): F85-104.
[22] ZHENG Y, ZHANG M, ZHAO Y, et al. JNK inhibitor SP600125 protects against lipopolysaccharide-induced acute lung injury via upregulation of claudin-4[J]. Exp Ther Med, 2014, 8(1): 153-158.
[23] LEE C Y, YANG J J, LEE S S, et al. Protective effect of Ginkgo biloba leaves extract, EGb761, on endotoxin-induced acute lung injury via a JNK- and Akt-dependent NFkappaB pathway[J]. J Agric Food Chem, 2014, 62(27): 6337-6344.
[24] WOLF P S, MERRY H E, FARIVAR A S, et al. Stress-activated protein kinase inhibition to ameliorate lung ischemia reperfusion injury[J]. J Thorac Cardiovasc Surg, 2008, 135(3): 656-665.