PAR-2在烟曲霉菌感染的人角膜上皮细胞中的作用

doi:10.6040/j.issn.1671-7554.0.2024.1010

摘要/Abstract

摘要： 目的研究蛋白酶激活受体-2(protease activated receptor-2, PAR-2)在烟曲霉菌感染的人永生化角膜上皮细胞(human corneal epithelial cells, HCECs)中的表达,探讨PAR-2对炎症因子水平的影响及其作用机制。方法烟曲霉菌灭活菌丝刺激HCECs,制备体外烟曲霉菌感染模型,观察烟曲霉菌灭活菌丝对PAR-2表达水平的影响,并观察PAR-2 拮抗剂(FSLLRY-NH₂)预处理HCECs后,炎症因子IL-1β和TNF-α,以及信号通路分子p-p38的水平变化。结果烟曲霉菌刺激HCECs后PAR-2表达水平升高。烟曲霉菌刺激引起IL-1β、TNF-α以及p-p38的表达升高,这一升高被PAR-2拮抗剂明显抑制。结论烟曲霉菌刺激增加PAR-2的表达,并通过PAR-2/p38 MAPK信号通路诱导炎症因子的产生。

关键词: 蛋白酶激活受体-2, 角膜上皮细胞, 烟曲霉菌, 炎症因子, 信号通路

Abstract: Objective To investigate the expression and function of protease activated receptor-2(PAR-2)in human corneal epithelial cells(HCECs)stimulated with Aspergillus fumigatus(A. fumigatus). Methods 75% ethanol-killed A. fumigatus stimulated HCECs, and PAR-2 mRNA and protein expressions were tested by RT-qPCR and Western blotting. HCECs were stimulated with 75% ethanol-killed A. fumigatus with or without pretreatment of FSLLRY-NH₂, PCR, Western blotting, and ELISA tested the expressions of IL-1β, TNF-α and p-p38. Results In HCECs, PAR-2 mRNA and protein expressions were significantly increased by A. fumigatus. A. fumigatus upregulated IL-1β, TNF-α and p-p38 expressions levels. FSLLRY-NH₂ significantly inhibited IL-1β, TNF-α and p-p38 expressions compared with infected control. Conclusion A. fumigates stimulation increases PAR-2 expression and upregulates cytokine expression levels though the PAR-2/p-p38 pathway.

Key words: Protease activated receptor-2, Corneal epithelial cells, Aspergillus fumigatus, Cytokines, Signal pathway

中图分类号:

R772.2

牛雅文,李凤娇. PAR-2在烟曲霉菌感染的人角膜上皮细胞中的作用[J]. 山东大学学报 (医学版), 2024, 62(12): 96-101.

NIU Yawen, LI Fengjiao. Role of PAR-2 during Aspergillus fumigatus infection in human corneal epithelial cells[J]. Journal of Shandong University (Health Sciences), 2024, 62(12): 96-101.

参考文献

[1] 庞金鼎, 韦振宇, 曹凯, 等. 不同抗真菌药物治疗真菌性角膜炎有效性和安全性的网状Meta分析[J]. 中华实验眼科杂志, 2024, 42(7): 629-637. PANG Jinding, WEI Zhenyu, CAO Kai, et al. A network meta-analysis of the efficacy and safety of different antifungal drugs for fungal keratitis[J]. Chinese Journal of Experimental Ophthalmology, 2024, 42(7): 629-637.
[2] 戴晨阳,郭慧. 白细胞介素-36在真菌性角膜炎中的免疫作用及机制[J]. 山东大学学报(医学版), 2024, 62(8): 67-73. DAI Chenyang, GUO Hui. Progress in the immune effect and mechanism of IL-36 in fungal keratitis[J]. Journal of Shandong University(Health Sciences), 2024, 62(8): 67-73.
[3] Brown L, Leck AK, Gichangi M, et al. The global incidence and diagnosis of fungal keratitis[J]. Lancet Infect Dis, 2021, 21(3): 49-57.
[4] 周晓丹, 杨玉倩, 徐强崧. 真菌性角膜炎的致病菌菌属和转归及其影响因素分析[J]. 国际眼科杂志, 2022, 22(11): 1892-1895. ZHOU Xiaodan, YANG Yuqian, XU Qiangsong. Analysis of pathogenic bacterial Genera and outcome of fungal keratitis and their influencing factors[J]. International Eye Science, 2022, 22(11): 1892-1895.
[5] Mahmoudi S, Masoomi A, Ahmadikia K, et al. Fungal keratitis: an overview of clinical and laboratory aspects[J]. Mycoses, 2018, 61(12): 916-930.
[6] Mills B, Radhakrishnan N, Karthikeyan Rajapandian SG, et al. The role of fungi in fungal keratitis[J]. Exp Eye Res, 2021, 202: 108372. doi:10.1016/j.exer.2020.108372.
[7] Heinekamp T, Schmidt H, Lapp K, et al. Interference of Aspergillus fumigatus with the immune response[J]. Semin Immunopathol, 2015, 37(2): 141-152.
[8] Shpacovitch V, Feld M, Bunnett NW, et al. Protease-activated receptors: novel PARtners in innate immunity[J].Trends Immunol, 2007, 28(12): 541-550.
[9] Moretti S, Bellocchio S, Bonifazi P, et al. The contribution of PARs to inflammation and immunity to fungi[J]. Mucosal Immunol, 2008, 1(2): 156-168.
[10] Rayees S, Joshi JC, Joshi B, et al. Protease-activated receptor 2 promotes clearance of Pseudomonas aeruginosa infection by inducing cAMP-Rac1 signaling in alveolar macrophages[J]. Front Pharmacol, 2022, 13: 874197. doi:10.3389/fphar.2022.874197.
[11] Lee SE, Kim JM, Jeong SK, et al. Protease-activated receptor-2 mediates the expression of inflammatory cytokines, antimicrobial peptides, and matrix metalloproteinases in keratinocytes in response to Propionibacterium acnes[J]. Arch Dermatol Res, 2010, 302(10): 745-756.
[12] Niu YW, Zhao GQ, Li C, et al. Aspergillus fumigatus increased PAR-2 expression and elevated proinflammatory cytokines expression through the pathway of PAR-2/ERK1/2 in Cornea[J]. Invest Ophthalmol Vis Sci, 2018, 59(1): 166-175.
[13] Rivas CM, Schiff HV, Moutal A, et al. Alternaria alternata-induced airway epithelial signaling and inflammatory responses via protease-activated receptor-2 expression[J]. Biochem Biophys Res Commun, 2022, 591: 13-19. doi:10.1016/j.bbrc.2021.12.090.
[14] Tripathi T, Abdi M, Alizadeh H. Protease-activated receptor 2(PAR2)is upregulated by Acanthamoeba plasminogen activator(aPA)and induces proinflammatory cytokine in human corneal epithelial cells[J]. Invest Ophthalmol Vis Sci, 2014, 55(6): 3912-3921.
[15] Wilson SE, Torricelli AAM, Marino GK. Corneal epithelial basement membrane: structure, function and regeneration[J]. Exp Eye Res, 2020, 194: 108002. doi:10.1016/j.exer.2020.108002.
[16] Chi MH, Gu LW, Zhang LN, et al. Pentoxifylline treats Aspergillus fumigatus keratitis by reducing fungal burden and suppressing corneal inflammation[J]. Eur J Pharmacol, 2023, 945: 175607. doi:10.1016/j.ejphar.2023.175607.
[17] Wang Ms J, Kang Ms X, Huang Ms ZQ, et al. Protease-activated receptor-2 decreased zonula occlidens-1 and claudin-1 expression and induced epithelial barrier dysfunction in allergic rhinitis[J]. Am J Rhinol Allergy, 2021, 35(1): 26-35.
[18] Matos NLA, Oliveira Lima OSC, DA Silva JF, et al. Blockade of protease-activated receptor 2 attenuates allergenmediated acute lung inflammation and leukocyte recruitment in mice[J]. J Biosci, 2022, 47: 2.
[19] Silva IS, Almeida AD, Lima Filho ACM, et al. Platelet-activating factor and protease-activated receptor 2 cooperate to promote neutrophil recruitment and lung inflammation through nuclear factor-kappa B transactivation[J]. Sci Rep, 2023, 13(1): 21637. doi:10.1038/s41598-023-48365-1.
[20] de Almeida AD, Silva IS, Fernandes-Braga W, et al. A role for mast cells and mast cell tryptase in driving neutrophil recruitment in LPS-induced lung inflammation via protease-activated receptor 2 in mice[J]. Inflamm Res, 2020, 69(10): 1059-1070.
[21] Chao HH, Chen PY, Hao WR, et al. Lipopolysaccharide pretreatment increases protease-activated receptor-2 expression and monocyte chemoattractant protein-1 secretion in vascular endothelial cells[J]. J Biomed Sci, 2017, 24(1): 85. doi:10.1186/s12929-017-0393-1.
[22] Cuadrado A, Nebreda AR. Mechanisms and functions of p38 MAPK signalling[J]. Biochem J, 2010, 429(3): 403-417.
[23] Wang JH, Liu YJ, Guo YS, et al. Function and inhibition of P38 MAP kinase signaling: targeting multiple inflammation diseases[J]. Biochem Pharmacol, 2024, 220: 115973. doi:10.1016/j.bcp.2023.115973.
[24] Sarg NH, Zaher DM, Abu Jayab NN, et al. The interplay of p38 MAPK signaling and mitochondrial metabolism, a dynamic target in cancer and pathological contexts[J]. Biochem Pharmacol, 2024, 225: 116307. doi:10.1016/j.bcp.2024.116307.
[25] Saleem S. Targeting MAPK signaling: a promising approach for treating inflammatory lung disease[J]. Pathol Res Pract, 2024, 254: 155122. doi:10.1016/j.prp.2024.155122.
[26] Niu YW, Lin J, Li C, et al. Galectin-3 plays an important pro-inflammatory role in A. fumigatus keratitis by recruiting neutrophils and activating p38 in neutrophils[J]. Int Immunopharmacol, 2021, 97: 107706. doi:10.1016/j.intimp.2021.107706.
[27] Wang Q, Zhao GQ, Lin J, et al. Role of the mannose receptor during Aspergillus fumigatus infection and interaction with dectin-1 in corneal epithelial cells[J]. Cornea, 2016, 35(2): 267-273.
[28] Diao WL, Yin M, Qi YH, et al. Resveratrol has neuroprotective effects and plays an anti-inflammatory role through Dectin-1/p38 pathway in Aspergillus fumigatus keratitis[J]. Cytokine, 2024, 179: 156626. doi:10.1016/j.cyto.2024.156626.
[29] Jia YY, Li C, Yin M, et al. Kaempferol ameliorate the prognosis of Aspergillus fumigatus keratitis by reducing fungal load and inhibiting the Dectin-1 and p38 MAPK pathway[J]. Exp Eye Res, 2022, 216: 108960. doi:10.1016/j.exer.2022.108960.

相关文章 15

[1]	潘静,于潇,刘金星,刘鹏飞. 始基卵泡激活调控的研究进展[J]. 山东大学学报 (医学版), 2024, 62(8): 74-92.
[2]	闫小龙,秦英,邵将,陈东峰,管东辉,赵灿斌. 姜黄素通过Wnt/β-catenin信号通路调控骨形成的机制[J]. 山东大学学报 (医学版), 2024, 62(10): 76-86.
[3]	秦金金,曹辰媛,邢杰杰,安燕,黄煜湘. 子痫前期相关Siglec-6核心基因的预测及生物信息学分析[J]. 山东大学学报 (医学版), 2024, 62(1): 31-37.
[4]	刘金波,刘凯文,向崇鑫,程雷. 西红花苷对椎间盘退变的保护作用[J]. 山东大学学报 (医学版), 2023, 61(9): 84-93.
[5]	张嘉颖,宿荣允,王英惠,王洪刚,柳刚. ACE2基因通过调控Nrf2/HO-1通路改善肾缺血再灌注损伤[J]. 山东大学学报 (医学版), 2023, 61(4): 1-9.
[6]	芦兴晨,强晔,刘昕宇,左丹,姜子晗,张玉超,刘元涛,马小莉. NR4A1对毒胡萝卜素诱导C2C12肌管细胞糖代谢功能障碍的影响[J]. 山东大学学报 (医学版), 2023, 61(11): 38-47.
[7]	邹品衡,陈添果,胡康,李伟才. 过表达miR-27a对急性脑梗死大鼠海马神经元损伤的影响及其机制[J]. 山东大学学报 (医学版), 2022, 60(9): 59-66.
[8]	张凤,吴哲,徐俊,刘玉兰. 6例非酒精性脂肪性肝病小鼠肠道B细胞的变化[J]. 山东大学学报 (医学版), 2022, 60(9): 67-73.
[9]	张秉芬,周胜红,王哲. 延龄草皂苷通过抑制TGF-β/Smad3与Wnt/β-catenin信号通路改善大鼠肺纤维化[J]. 山东大学学报 (医学版), 2022, 60(8): 23-29.
[10]	相宇娇,刘强,刘璐,石艳. 原发免疫性血小板减少症树突状细胞异常免疫反应机制[J]. 山东大学学报 (医学版), 2022, 60(7): 89-97.
[11]	邬雨洁,张明泉,纪永利,赵璐,王越,陈沙沙. 寒痉汤及其拆方对寒凝证高血压大鼠血清炎症因子、血管内皮功能及纤维化的影响[J]. 山东大学学报 (医学版), 2022, 60(6): 10-18.
[12]	张秀芳,李沛铮,张博涵,孙丛丛,刘艺鸣. 生长分化因子15在LPS诱导的帕金森病模型中的保护作用及机制[J]. 山东大学学报 (医学版), 2022, 60(5): 1-7.
[13]	虎娜,孙苗,邢莎莎,许丹霞,海小明,马玲,杨丽,勉昱琛,何瑞,陈冬梅,马会明. 月见草油抵抗多囊卵巢综合征大鼠卵巢氧化应激[J]. 山东大学学报 (医学版), 2022, 60(5): 22-30.
[14]	申晓畅,孙一卿,颜磊,赵兴波. 芳基烃受体核转位因子样蛋白2在子宫内膜癌中的表达[J]. 山东大学学报 (医学版), 2022, 60(5): 74-80.
[15]	孙继业,王紫欧,孙晓伟,李洪涛. 中药熏蒸联合体外冲击波对72例髋关节撞击综合征临床疗效、血清炎症因子水平的影响[J]. 山东大学学报 (医学版), 2022, 60(4): 76-81.

多维度评价

Viewed

Full text

Abstract

Cited

Shared

Discussed