细颗粒物通过上调毒蕈碱M3受体诱导小鼠气道高反应性  

Fine particulate matter induces airway hyper-responsiveness by up-regulating muscarinic M_(3) receptor in mice

作　　者：王荣[1] 王娜娜[1] 杨宽[1] 余丽丽[1] 秦蓓[1] WANG Rong;WANG Nana;YANG Kuan;YU Lili;QIN Bei(Xi'an Key Laboratory for Research and Development of Innovative Multi-Target Antihypertensive Drugs,Xi'an Innovative Antihypertensive Drugs International Science and Technology Cooperation Base,Institute of Drug Research,Xi'an Medical University,College of Pharmacy,Xi'an Medical University,Xi'an 710021,China)

机构地区：[1]西安市多靶协同抗高血压创新药物研制重点实验室,西安市智创抗高血压药物国际科技合作基地,西安医学院药物研究所,西安医学院药学院,陕西西安710021

出　　处：《中国病理生理杂志》2025年第4期696-703,共8页Chinese Journal of Pathophysiology

基　　金：陕西省重点研发计划项目(No.2021ZDLSF03-05);国家自然科学基金资助项目(No.81903288);西安医学院校级科技创新团队(No.2021TD03、07);陕西省教育厅科研计划项目(No.23JP155);教育部产学合作协同育人项目(No.2024CXHZ-03)。

摘　　要：目的:细颗粒物(fine particulate matter,PM_(2.5))与气道高反应性(airway hyper-responsiveness,AHR)密切相关,然而,PM_(2.5)导致AHR的潜在机制仍不清楚。本研究旨在探讨大气PM_(2.5)暴露对诱导AHR的可能作用机理。方法:小鼠40只随机分为生理盐水对照组、脂多糖(lipopolysaccharide,LPS)组(100 mg/L)、PM_(2.5)低剂量组(0.0035 mg/d)、PM_(2.5)中剂量组(0.007 mg/d)和PM_(2.5)高剂量组(0.014 mg/d),PM_(2.5)鼻内滴注连续30 d。清醒动物肺功能测量系统和离体肌张力测定仪评价肺功能和气管收缩反应。酶联免疫吸附测定法(enzyme-linked immunosorbent assay,ELISA)检测血清中炎症介质和氧化应激指标。苏木精-伊红(hematoxylin and eosin,HE)和Masson染色检测小鼠肺组织病理变化。实时荧光定量PCR(quantitative real-time PCR,RT-qPCR)和Western blot检测气道收缩性受体及丝裂原活化蛋白激酶(mitogen-activated protein kinase,MAPK)信号通路的活化情况。结果:PM_(2.5)滴鼻可增加小鼠的气道阻力,增强毒蕈碱受体(muscarine receptor,M受体)介导的气管收缩。PM_(2.5)滴鼻可增加血清中肿瘤坏死因子α(tumor necrosis factor-α,TNF-α)、白细胞介素1β(interleukin-1β,IL-1β)和IL-6的含量。鼻内滴入PM_(2.5)可显著降低谷胱甘肽过氧化物酶(glutathione peroxidase,GSH-Px)水平,增加丙二醛(malondialdehyde,MDA)水平。HE和Masson染色结果显示,PM_(2.5)滴鼻造成炎性细胞浸润和肺泡上皮细胞增生等明显的肺组织病理变化,并且促进胶原沉积。PM_(2.5)暴露增加了M_(3)受体mRNA和蛋白的表达,增加了p-ERK1/2和p-p38的表达。结论:PM_(2.5)滴鼻可引起炎症反应和氧化应激,激活细胞外调节蛋白激酶1/2(extracellular signal-regulated kinase 1/2,ERK1/2)和p38MAPK通路,上调M_(3)受体,诱导气道反应性。AIM:Fine particulate matter(PM_(2.5))is closely associated with airway hyper-responsiveness(AHR).However,the underlying mechanism by which PM_(2.5) leads to AHR is still unclear.This study aimed to investigate the respiratory effects of ambient PM_(2.5) exposure.METHODS:Forty mice were randomly divided into five groups:control group(intranasal saline),lipopolysaccharide(LPS)group(100 mg/L),PM_(2.5) low-dose group(0.0035 mg/d),PM_(2.5) medium-dose group(0.007 mg/d),and PM_(2.5) high-dose group(0.014 mg/d).They were treated with intranasal instillation for 30 d.Lung function and tracheal contractile responses were evaluated using whole-body plethysmography and sensitive wire myograph.Inflammatory mediators in serum and oxidative stress parameters were detected using enzymelinked immunosorbent assay(ELISA).Lung tissues were subjected to HE and Masson staining.RT-qPCR and Western blot were used to determine the expression of contractile receptors and the phosphorylation of the mitogen-activated protein kinase(MAPK)signal pathway.RESULTS:Intranasal instillation of PM_(2.5) significantly increased airway resistance in mice and enhanced tracheal contractility in response to carbachol.PM_(2.5) elevated levels of tumor necrosis factor-α(TNF-α),interleukin(IL)-1β,and IL-6 in serum.PM_(2.5) instillation also led to a decrease in glutathione peroxidase(GSH-Px)levels and an increase in malondialdehyde(MDA)levels.Lung tissue exhibited notable pathological changes,including inflammatory cell infiltration,hyperplasia of alveolar epithelial cells,and collagen deposition.Mechanistically,exposure to PM_(2.5) increased the expression of muscarinic M_(3) receptor mRNA and protein,as well as the phosphorylation of p-ERK1/2 and p-p38 proteins following PM_(2.5) instillation.CONCLUSION:Intranasal instillation of PM_(2.5) induced inflammation and oxidative stress,along with the activation of the extracellular signal-regulated kinase 1/2(ERK1/2)and p38 MAPK pathways,resulting in the upregulation of M_(3) receptor-induced AHR.

关 键 词：细颗粒物 气道高反应性 毒蕈碱受体 MAPK信号通路 

分 类 号：R563[医药卫生—呼吸系统] X513[医药卫生—内科学] R363.2[医药卫生—临床医学]