The Journal of Practical Medicine >
House dust mite⁃induced autophagy affects airway epithelial barrier function through β⁃catenin⁃Snail signaling pathway
Received date: 2025-02-07
Online published: 2025-05-20
Objective To investigate the mechanism of autophagy induced by House dust mites （HDM） on airway epithelial tight junction through β-catenin-Snail signaling pathway. Methods Human bronchial epithelial cells （16HBE） were stimulated with HDM at different time points （0， 3， 6， 12， 24， 48 h） and different concentrations （0， 40， 100， 200 μg/mL） to screen the appropriate stimulation concentration and stimulation time. 16HBE cells were treated with oxidative stress inhibitor N-acetylcysteine （NAC）， autophagy inhibitor 3-methyladenine （3-MA）， HDM， and their combinations. Cells were transfected with mCherry-EGFP-LC3B， Beclin-1-siRNA， and ATG14-siRNA lentivirus and then stimulated with NAC and HDM. Immunofluorescence was used to detect the expression levels of autophagy-related protein LC3B， tight junction-related proteins Occludin， and ZO-1 in airway epithelial cells. The level of reactive oxygen species （ROS） was detected by using DCFH-DA in each group. The protein expression levels of Occludin， ZO-1， LC3B， Beclin-1， ATG5， ATG14， P62， Snail， β-catenin and p-β-catenin were detected by Western blot method. Results Immunofluorescence results showed that compared with the control group， 200 μg/mL HDM stimulation induced cellular autophagy， increased the expression level of LC3B protein， and promoted the level of ROS， all with statistical significances（all P < 0.05）. Compared with the HDM group， the HDM + 3-MA， HDM + ATG14-si， and HDM + Beclin-1-si groupsall showed significantincreases in the expression levels of tight junction-related proteins Occludin and ZO-1 （P < 0.05）. The HDM + NAC group demonstrated significant decreases both in the level of ROS andin the expression level of LC3B protein.Western blot results revealed that compared with HDM， 3-MA and autophagy protein low-expression beads （Beclin-1-si， ATG14-si） attenuated HDM-induced cellular autophagy （P < 0.05）， inhibited HDM-induced upregulation of Snail and p-β- catenin expression， and improved HDM-induced decreases in Occludin and ZO-1（P < 0.05）. Moreover， compared with the HDM group， the NAC + HDM group exhibited significant decreases both in the conversion of LC3BⅠ to LC3BⅡ （P < 0.001） in the protein levels of Snail， p-β-catenin， Beclin-1 and ATG14 （P < 0.01）， but significant increases in the protein levels of Occludin and ZO-1 （P < 0.05）. Conclusion HDM affects the tight connections between airway epithelial cells by inducing autophagy， which may be attributed to the β-catenin-Snail signaling pathway.
Ziling ZENG , Xing WANG , Hongmei TANG , Zhibin WANG , Ning MA , Yuejiao LI , Xiaoyun WANG , Xiefang YUAN , Guofeng XU , Qiaoqiao WANG , Wen ZHANG , Jiayao DUAN , Yun ZHANG . House dust mite⁃induced autophagy affects airway epithelial barrier function through β⁃catenin⁃Snail signaling pathway[J]. The Journal of Practical Medicine, 2025 , 41(9) : 1309 -1318 . DOI: 10.3969/j.issn.1006-5725.2025.09.006
| 1 | HE Z， FENG J， XIA J， et al. Frequency of Signs and Symptoms in Persons With Asthma ［J］. Respir Care， 2020， 65（2）： 252-264. doi:10.4187/respcare.06714 |
| 2 | WANG J Y. The innate immune response in house dust mite-induced allergic inflammation ［J］. Allergy Asthma Immunol Res， 2013， 5（2）： 68-74. doi:10.4168/aair.2013.5.2.68 |
| 3 | STEVENSON C S， BIRRELL M A. Moving towards a new generation of animal models for asthma and COPD with improved clinical relevance ［J］. Pharmacol Ther， 2011， 130（2）： 93-105. doi:10.1016/j.pharmthera.2010.10.008 |
| 4 | WANG Q， GUO L， ZENG Z， et al. Neferine Attenuates HDM-Induced Allergic Inflammation by Inhibiting the Activation of Dendritic Cell ［J］. Inflammation， 2023， 46（6）： 2433-2448. doi:10.1007/s10753-023-01891-6 |
| 5 | GON Y， HASHIMOTO S. Role of airway epithelial barrier dysfunction in pathogenesis of asthma ［J］. Allergol Int， 2018， 67（1）： 12-17. doi:10.1016/j.alit.2017.08.011 |
| 6 | ZIHNI C， MILLS C， MATTER K， et al. Tight junctions： from simple barriers to multifunctional molecular gates ［J］. Nat Rev Mol Cell Biol， 2016， 17（9）： 564-580. doi:10.1038/nrm.2016.80 |
| 7 | GEORAS S N， REZAEE F. Epithelial barrier function： At the front line of asthma immunology and allergic airway inflammation ［J］. J Allergy Clin Immunol， 2014， 134（3）： 509-520. doi:10.1016/j.jaci.2014.05.049 |
| 8 | PARK S W， LEE E H， LEE E J， et al. Apolipoprotein A1 potentiates lipoxin A4 synthesis and recovery of allergen-induced disrupted tight junctions in the airway epithelium ［J］. Clin Exp Allergy， 2013， 43（8）： 914-927. doi:10.1111/cea.12143 |
| 9 | GANAPATHY A S， SAHA K， SUCHANEC E， et al. AP2M1 mediates autophagy-induced CLDN2 （claudin 2） degradation through endocytosis and interaction with LC3 and reduces intestinal epithelial tight junction permeability ［J］. Autophagy， 2022， 18（9）： 2086-2103. doi:10.1080/15548627.2021.2016233 |
| 10 | LIU M， WANG Q， WU W， et al. Glaesserella parasuis serotype 5 breaches the porcine respiratory epithelial barrier by inducing autophagy and blocking the cell membrane Claudin-1 replenishment ［J］. PLoS Pathog， 2022， 18（10）： e1010912. doi:10.1371/journal.ppat.1010912 |
| 11 | ZHOU J S， ZHAO Y， ZHOU H B， et al. Autophagy plays an essential role in cigarette smoke-induced expression of MUC5AC in airway epithelium ［J］. Am J Physiol Lung Cell Mol Physiol， 2016， 310（11）： L1042-1052. doi:10.1152/ajplung.00418.2015 |
| 12 | DICKINSON J D， ALEVY Y， MALVIN N P， et al. IL13 activates autophagy to regulate secretion in airway epithelial cells ［J］. Autophagy， 2016， 12（2）： 397-409. doi:10.1080/15548627.2015.1056967 |
| 13 | CHEN Z H， WU Y F， WANG P L， et al. Autophagy is essential for ultrafine particle-induced inflammation and mucus hyperproduction in airway epithelium ［J］. Autophagy， 2016， 12（2）： 297-311. doi:10.1080/15548627.2015.1124224 |
| 14 | FAN Y， HOU T， DAN W， et al. Silibinin inhibits epithelial? mesenchymal transition of renal cell carcinoma through autophagy?dependent Wnt/β?catenin signaling ［J］. Int J Mol Med， 2020， 45（5）： 1341-1350. |
| 15 | TIAN Y， WU J， ZENG L， et al. Huaier polysaccharides suppress triple-negative breast cancer metastasis and epithelial-mesenchymal transition by inducing autophagic degradation of Snail ［J］. Cell Biosci， 2021， 11（1）： 170. doi:10.1186/s13578-021-00682-6 |
| 16 | YU F， YU C， LI F， et al. Wnt/β-catenin signaling in cancers and targeted therapies ［J］. Signal Transduct Target Ther， 2021， 6（1）： 307. doi:10.1038/s41392-021-00701-5 |
| 17 | LI J， LIU N， ZHOU H， et al. Immunoproteasome inhibition prevents progression of castration-resistant prostate cancer ［J］. Br J Cancer， 2023， 128（7）： 1377-1390. doi:10.1038/s41416-022-02129-2 |
| 18 | KANLAYA R， KAPINCHARANON C， FONG-NGERN K， et al. Induction of mesenchymal-epithelial transition （MET） by epigallocatechin-3-gallate to reverse epithelial-mesenchymal transition （EMT） in SNAI1-overexpressed renal cells： A potential anti-fibrotic strategy ［J］. J Nutr Biochem， 2022， 107： 109066. doi:10.1016/j.jnutbio.2022.109066 |
| 19 | GREGORY L G， LLOYD C M. Orchestrating house dust mite-associated allergy in the lung ［J］. Trends Immunol， 2011， 32（9）： 402-411. doi:10.1016/j.it.2011.06.006 |
| 20 | ZEKI A A， YEGANEH B， KENYON N J， et al. Autophagy in airway diseases： A new frontier in human asthma？ ［J］. Allergy， 2016， 71（1）： 5-14. doi:10.1111/all.12761 |
| 21 | RACANELLI A C， KIKKERS S A， CHOI A M K， et al. Autophagy and inflammation in chronic respiratory disease ［J］. Autophagy， 2018， 14（2）： 221-232. doi:10.1080/15548627.2017.1389823 |
| 22 | HE C， KLIONSKY D J. Regulation mechanisms and signaling pathways of autophagy ［J］. Annu Rev Genet， 2009， 43： 67-93. doi:10.1146/annurev-genet-102808-114910 |
| 23 | ALBANO G D， GAGLIARDO R P， MONTALBANO A M， et al. Overview of the Mechanisms of Oxidative Stress： Impact in Inflammation of the Airway Diseases ［J］. Antioxidants （Basel）， 2022， 11（11）： 2237. doi:10.3390/antiox11112237 |
| 24 | SUZUKI Y， AONO Y， AKIYAMA N， et al. Involvement of autophagy in exacerbation of eosinophilic airway inflammation in a murine model of obese asthma ［J］. Autophagy， 2022， 18（9）： 2216-2228. doi:10.1080/15548627.2022.2025571 |
| 25 | 颜鹏， 邓育琼， 黄杏兰， 等. 地塞米松对哮喘模型小鼠肺组织活性氧及线粒体基因MTCO1的影响 ［J］. 实用医学杂志， 2022， 38（6）： 731-737. |
| 26 | FILOMENI G， DE ZIO D， CECCONI F. Oxidative stress and autophagy： The clash between damage and metabolic needs ［J］. Cell Death Differ， 2015， 22（3）： 377-388. doi:10.1038/cdd.2014.150 |
| 27 | QIU Y N， WANG G H， ZHOU F， et al. PM2.5 induces liver fibrosis via triggering ROS-mediated mitophagy ［J］. Ecotoxicol Environ Saf， 2019， 167： 178-187. doi:10.1016/j.ecoenv.2018.08.050 |
| 28 | WANG B， WANG Y， ZHANG J， et al. ROS-induced lipid peroxidation modulates cell death outcome： Mechanisms behind apoptosis， autophagy， and ferroptosis ［J］. Arch Toxicol， 2023， 97（6）： 1439-1451. doi:10.1007/s00204-023-03476-6 |
| 29 | KIM N Y， SHIVANNE GOWDA S G， LEE S G， et al. Cannabidiol induces ERK activation and ROS production to promote autophagy and ferroptosis in glioblastoma cells ［J］. Chem Biol Interact， 2024， 394： 110995. doi:10.1016/j.cbi.2024.110995 |
| 30 | RABY K L， MICHAELOUDES C， TONKIN J， et al. Mechanisms of airway epithelial injury and abnormal repair in asthma and COPD ［J］. Front Immunol， 2023， 14： 1201658. doi:10.3389/fimmu.2023.1201658 |
| 31 | HUANG Z Q， LIU J， SUN L Y， et al. Updated epithelial barrier dysfunction in chronic rhinosinusitis： Targeting pathophysiology and treatment response of tight junctions ［J］. Allergy， 2024， 79（5）： 1146-1165. doi:10.1111/all.16064 |
| 32 | STEELANT B， FARRé R， WAWRZYNIAK P， et al. Impaired barrier function in patients with house dust mite-induced allergic rhinitis is accompanied by decreased occludin and zonula occludens-1 expression ［J］. J Allergy Clin Immunol， 2016， 137（4）： 1043-1053.e1045. doi:10.1016/j.jaci.2015.10.050 |
| 33 | VINCENT T， NEVE E P， JOHNSON J R， et al. A SNAIL1-SMAD3/4 transcriptional repressor complex promotes TGF-beta mediated epithelial-mesenchymal transition ［J］. Nat Cell Biol， 2009， 11（8）： 943-950. doi:10.1038/ncb1905 |
| 34 | ZHANG X， YU X， JIANG G， et al. Cytosolic TMEM88 promotes invasion and metastasis in lung cancer cells by binding DVLS ［J］. Cancer Res， 2015， 75（21）： 4527-4537. doi:10.1158/0008-5472.can-14-3828 |
| 35 | KYUNO D， KOJIMA T， YAMAGUCHI H， et al. Protein kinase Cα inhibitor protects against downregulation of claudin-1 during epithelial-mesenchymal transition of pancreatic cancer ［J］. Carcinogenesis， 2013， 34（6）： 1232-1243. doi:10.1093/carcin/bgt057 |
| 36 | MARTíNEZ-ESTRADA O M， LETTICE L A， ESSAFI A， et al. Wt1 is required for cardiovascular progenitor cell formation through transcriptional control of Snail and E-cadherin ［J］. Nat Genet， 2010， 42（1）： 89-93. doi:10.1038/ng.494 |
| 37 | GNEMMI V， BOUILLEZ A， GAUDELOT K， et al. MUC1 drives epithelial-mesenchymal transition in renal carcinoma through Wnt/β-catenin pathway and interaction with SNAIL promoter ［J］. Cancer Lett， 2014， 346（2）： 225-236. doi:10.1016/j.canlet.2013.12.029 |
| 38 | 肖知周， 黄莺， 卞华伟. 基于Wnt/β-catenin信号通路介导的自噬和凋亡探讨苦豆子碱对骨质疏松小鼠骨代谢的影响 ［J］. 实用医学杂志， 2025， 41（4）： 500-508. |
| 39 | DAI X M， ZHANG Y H， LIN X H， et al. SIK2 represses AKT/GSK3β/β-catenin signaling and suppresses gastric cancer by inhibiting autophagic degradation of protein phosphatases ［J］. Mol Oncol， 2021， 15（1）： 228-245. doi:10.1002/1878-0261.12838 |
| 40 | OVERHOFF M， TELLKAMP F， HESS S， et al. Autophagy regulates neuronal excitability by controlling cAMP/protein kinase A signaling at the synapse ［J］. EMBO J， 2022， 41（22）： e110963. doi:10.15252/embj.2022110963 |
| 41 | CHENG L， XU Y， LONG Y， et al. Liraglutide attenuates palmitate-induced apoptosis via PKA/β-catenin/Bcl-2/Bax pathway in MC3T3-E1 cells ［J］. Naunyn Schmiedebergs Arch Pharmacol， 2024， 397（1）： 329-341. doi:10.1007/s00210-023-02572-9 |
/
| 〈 |
|
〉 |