屋尘螨诱导的自噬通过β-catenin-Snail信号通路影响气道上皮屏障功能

doi:10.3969/j.issn.1006-5725.2025.09.006

Abstract

Abstract:

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.

Key words: house dust mite, cellular autophagy, human bronchial epithelial cells, tight junctions

CLC Number:

R563

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.

Figures/Tables 5

References 41

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 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 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 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 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 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 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 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 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 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 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 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 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 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 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 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 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 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 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 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 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 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 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 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 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 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 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 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 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 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 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 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 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 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 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 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 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 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 doi: 10.1007/s00210-023-02572-9

[1]	Fuxia ZHENG,Lijun MIAO,Fengxiang HUANG,Shifu HUANG,Zengyan GAO,Ruixia ZHANG,Yong MENG. The correlation between the levels of APC， TXB2， and sB7⁃H3 in peripheral blood of elderly patients with pneumonia and the severity and prognosis of the disease [J]. The Journal of Practical Medicine, 2025, 41(7): 1056-1061.
[2]	Yanqing MAO,Ting FU,Juan WANG,Ya′nan WANG,Jie. LI. Expression levels of miR⁃127⁃5p， miR⁃127 and miR⁃155 in elderly patients with pneumonia infection by multidrug⁃resistant bacteria and their correlation with immune⁃related cytokines [J]. The Journal of Practical Medicine, 2025, 41(3): 385-390.
[3]	Wenfang HE,Xun ZHOU,Meizhu DING,Yiping CHEN,Qianli LIU,Xinyuan. TAN. Application of the "ASK⁃T" aspiration prevention management model in reducing the risk of aspiration in elderly patients with chronic obstructive pulmonary disease （COPD） complicated by dysphagia [J]. The Journal of Practical Medicine, 2025, 41(3): 434-441.
[4]	Yan JIANG,Xiaoqin WANG,Hong MEI,Xinxin LIU,Zhenliang LIAO,Kun YU,Banghai FENG,Song QIN. Type Ⅱ alveolar epithelial cell⁃derived exosomal miR⁃21⁃5p targeting SKP2 alleviate bronchopulmonary dysplasia [J]. The Journal of Practical Medicine, 2024, 40(23): 3298-3305.
[5]	Xinpan LI,Yi FANG,Jun QIU. Predictive value of CD3⁺/CD4⁺T lymphocyte level and neutrophil to lymphocyte ratio in radiation pneumonia [J]. The Journal of Practical Medicine, 2024, 40(20): 2923-2928.
[6]	Ling TAN,Xue MA,Jingjing WANG,Fengfeng LEI. Research progress of matrix metalloproteinase⁃28 in pulmonary disease [J]. The Journal of Practical Medicine, 2024, 40(20): 2949-2953.
[7]	Shengfang YUAN,Bu WANG,Baoli XIANG,Jianqing ZHAO,Jingjing SHEN,Zhihua. ZHANG. Prediction of immune therapy efficacy and prognosis for advanced non-small cell lung cancer using peripheral blood circulation tumor DNA [J]. The Journal of Practical Medicine, 2024, 40(15): 2110-2115.
[8]	Guomin ZHAO,Hui ZHANG,Pucong YE,Wei. CHEN. Effect of lactate dehydrogenase to albumin ratio on the short⁃term prognosis in patients with sepsis⁃associated acute kidney injury [J]. The Journal of Practical Medicine, 2024, 40(13): 1803-1807.
[9]	Yanqing MAO,Ya′nan WANG,Jie. LI. The clinical symptoms， quality of life and laboratory indexes of newly diagnosed chronic obstructive pulmonary disease with hypertension [J]. The Journal of Practical Medicine, 2024, 40(11): 1549-1553.
[10]	Shifang YANG,Xinglin GAO,Jing LI,Lupeng. JI. Sarcopenia associated with chronic obstructive pulmonary disease [J]. The Journal of Practical Medicine, 2024, 40(9): 1181-1185.
[11]	Ling XIAO,Chunlei GAO,Wei GUO,Ning WANG,Xuan ZHANG,Ming. LIU. Codonopsis polysaccharide protected LPS⁃induced acute lung injury by inhibiting MAPK/NF⁃κB signaling pathway in mice [J]. The Journal of Practical Medicine, 2024, 40(7): 948-954.
[12]	Jingcai WANG,Chunyan GUO,Lixin YANG,Xiaoqing. JING. Correlation of serum RAGE and HMGB1 expression with the occurrence of acute respiratory distress syndrome and IFN⁃γ/IL⁃4 ratio in patients with severe pneumonia [J]. The Journal of Practical Medicine, 2024, 40(4): 515-520.
[13]	Aiqin LI,Zhen ZHANG,Ya′nan XU,Jinyuan ZHU,Xu. ZHANG. Research progress on the interaction between macro⁃phages and fibroblasts in ARDS pulmonary fibrosis [J]. The Journal of Practical Medicine, 2024, 40(4): 571-574.
[14]	Lihong DING,Shijia GENG,Yujie WANG. Effects of wedelobata on apoptosis and secretion of inflammatory cytokines in the alveolar epithelium infected by Streptococcus pneumonia [J]. The Journal of Practical Medicine, 2024, 40(3): 316-320.
[15]	Changzhi ZHANG,Yuzhi TAO,Qian YU,Xunping WU,Weijia LIU,Jing HAN. Clinical efficacy evaluation and analysis of different acute pulmonary embolism prognostic scores [J]. The Journal of Practical Medicine, 2024, 40(3): 336-342.

House dust mite⁃induced autophagy affects airway epithelial barrier function through β⁃catenin⁃Snail signaling pathway

RichHTML

PDF (PC)

Knowledge

Abstract

Cite this article

share this article

Figures/Tables 5

References 41

Related Articles 15

Recommended Articles

Metrics

Comments