Silencing a disintegrin and metalloproteinase‑33 attenuates the proliferation of vascular smooth muscle cells via PI3K/AKT pathway: Implications in the pathogenesis of airway vascular remodeling

Yan,Fang; Hao,Yanyan; Gong,Xinji; Sun,Hu; Ding,Jianbing; Wang,Jing

doi:10.3892/mmr.2021.12141

Silencing a disintegrin and metalloproteinase‑33 attenuates the proliferation of vascular smooth muscle cells via PI3K/AKT pathway: Implications in the pathogenesis of airway vascular remodeling

Authors:
- Fang Yan
- Yanyan Hao
- Xinji Gong
- Hu Sun
- Jianbing Ding
- Jing Wang
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Affiliations: School of Public Health, Xinjiang Medical University, Urumqi, Xinjiang Uyghur Autonomous Region 830011, P.R. China, State Key Laboratory of Pathogenesis, Prevention and Treatment of High Incidence Diseases in Central Asia, Department of Respiratory Medicine, The First Affiliated Hospital of Xinjiang Medical University, Urumqi, Xinjiang Uyghur Autonomous Region 830054, P.R. China, Department of Immunology, College of Basic Medicine, Xinjiang Medical University, Urumqi, Xinjiang Uyghur Autonomous Region 830011, P.R. China
Published online on: May 9, 2021 https://doi.org/10.3892/mmr.2021.12141
Article Number: 502
Copyright: © Yan et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

Accumulating evidence suggests that pulmonary expression of a disintegrin and metalloproteinase‑33 (ADAM33) serves a key role in the pathogenesis of airway remodeling‑related diseases, including asthma. Airway vascular proliferation has been recognized as a key feature of airway remodeling. Our previous study showed that ADAM33 is constitutively expressed in airway vascular smooth muscle cells in patients with asthma, suggesting a potential role of ADAM33 in regulating airway vascular remodeling. Using in vitro human aortic smooth muscle cells (HASMCs) and lentiviral vector carrying short hairpin RNA for ADAM33, the present study aimed to evaluate the influence of ADAM33 silencing on the proliferation and apoptosis of HASMCs and the underlying molecular pathways. Cellular proliferation was observed using the Cell Counting Kit‑8 method. Cellular apoptosis was evaluated with Annexin V‑PE/7‑AAD staining and flow cytometry. Reverse transcription‑quantitative PCR and western blotting were used to evaluate the changes in mRNA and protein levels of involved signaling molecules. It was found that silencing of ADAM33 expression in HASMCs significantly inhibited proliferation, but induced the apoptosis of HASMCs. These changes were accompanied by inhibition of the PI3K/AKT/ERK pathway and Bcl‑2, but an increase in Bax expression. These results suggested that constitutive expression of ADAM33 may be important to maintain a proliferative phenotype in HASMCs. The influences of ADAM33 on proliferation and apoptosis of HASMCs may involve regulation of PI3K/AKT/ERK and Bax/Bcl‑2 pathways. These findings suggested an important role of ADAM33 in airway vascular remodeling and potential therapeutic significance of ADAM33 inhibition in airway remodeling‑related diseases.

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1	Hough KP, Curtiss ML, Blain TJ, Liu RM, Trevor J, Deshane JS and Thannickal VJ: Airway remodeling in asthma. Front Med (Lausanne). 7:1912020. View Article : Google Scholar : PubMed/NCBI
2	Kardas G, Kuna P and Panek M: Biological therapies of severe asthma and their possible effects on airway remodeling. Front Immunol. 11:11342020. View Article : Google Scholar : PubMed/NCBI
3	Fehrenbach H, Wagner C and Wegmann M: Airway remodeling in asthma: What really matters. Cell Tissue Res. 367:551–569. 2017. View Article : Google Scholar : PubMed/NCBI
4	Zhou-Suckow Z, Duerr J, Hagner M, Agrawal R and Mall MA: Airway mucus, inflammation and remodeling: Emerging links in the pathogenesis of chronic lung diseases. Cell Tissue Res. 367:537–550. 2017. View Article : Google Scholar : PubMed/NCBI
5	Harkness LM, Kanabar V, Sharma HS, Westergren-Thorsson G and Larsson-Callerfelt AK: Pulmonary vascular changes in asthma and COPD. Pulm Pharmacol Ther. 29:144–155. 2014. View Article : Google Scholar : PubMed/NCBI
6	Olivieri D and Chetta A: Therapeutic perspectives in vascular remodeling in asthma and chronic obstructive pulmonary disease. Chem Immunol Allergy. 99:216–225. 2014. View Article : Google Scholar : PubMed/NCBI
7	Barbato A, Turato G, Baraldo S, Bazzan E, Calabrese F, Panizzolo C, Zanin ME, Zuin R, Maestrelli P, Fabbri LM and Saetta M: Epithelial damage and angiogenesis in the airways of children with asthma. Am J Respir Crit Care Med. 174:975–981. 2006. View Article : Google Scholar : PubMed/NCBI
8	Alagappan VK, de Boer WI, Misra VK, Mooi WJ and Sharma HS: Angiogenesis and vascular remodeling in chronic airway diseases. Cell Biochem Biophys. 67:219–234. 2013. View Article : Google Scholar : PubMed/NCBI
9	Bakakos P, Patentalakis G and Papi A: Vascular biomarkers in asthma and COPD. Curr Top Med Chem. 16:1599–1609. 2016. View Article : Google Scholar : PubMed/NCBI
10	Rzucidlo EM: Signaling pathways regulating vascular smooth muscle cell differentiation. Vascular. 17 (Suppl 1):S15–S20. 2009. View Article : Google Scholar : PubMed/NCBI
11	Klein T and Bischoff R: Active metalloproteases of the A Disintegrin and Metalloprotease (ADAM) family: Biological function and structure. J Proteome Res. 10:17–33. 2011. View Article : Google Scholar : PubMed/NCBI
12	Dreymueller D, Uhlig S and Ludwig A: ADAM-family metalloproteinases in lung inflammation: Potential therapeutic targets. Am J Physiol Lung Cell Mol Physiol. 308:L325–L343. 2015. View Article : Google Scholar : PubMed/NCBI
13	Li HF, Yan LP, Wang K, Li XT, Liu HX and Tan W: Association between ADAM33 polymorphisms and asthma risk: A systematic review and meta-analysis. Respir Res. 20:382019. View Article : Google Scholar : PubMed/NCBI
14	Lee JY, Park SW, Chang HK, Kim HY, Rhim T, Lee JH, Jang AS, Koh ES and Park CS: A disintegrin and metalloproteinase 33 protein in patients with asthma: Relevance to airflow limitation. Am J Respir Crit Care Med. 173:729–735. 2006. View Article : Google Scholar : PubMed/NCBI
15	Poon AH, Houseman EA, Ryan L, Sparrow D, Vokonas PS and Litonjua AA: Variants of asthma and chronic obstructive pulmonary disease genes and lung function decline in aging. J Gerontol A Biol Sci Med Sci. 69:907–913. 2014. View Article : Google Scholar : PubMed/NCBI
16	Hur GY and Broide DH: Genes and pathways regulating decline in lung function and airway remodeling in asthma. Allergy Asthma Immunol Res. 11:604–621. 2019. View Article : Google Scholar : PubMed/NCBI
17	Kozlik P, Zuk J, Bartyzel S, Zarychta J, Okon K, Zareba L, Bazan JG, Kosalka J, Soja J, Musial J and Bazan-Socha S: The relationship of airway structural changes to blood and bronchoalveolar lavage biomarkers, and lung function abnormalities in asthma. Clin Exp Allergy. 50:15–28. 2020. View Article : Google Scholar : PubMed/NCBI
18	Zhang L, Hi X, Yao L, J. W and W. Q: The effect of interleukin4 on ADAM33 gene expression in airway wall vascular smooth muscle cells. Journal of Xinjiang Medical University. 39:265–269. 2016.PubMed/NCBI
19	Isenovic ER, Kedees MH, Tepavcevic S, Milosavljevic T, Koricanac G, Trpkovic A and Marche P: Role of PI3K/AKT, cPLA2 and ERK1/2 signaling pathways in insulin regulation of vascular smooth muscle cells proliferation. Cardiovasc Hematol Disord Drug Targets. 9:172–180. 2009. View Article : Google Scholar : PubMed/NCBI
20	Chae IH, Park KW, Kim HS and Oh BH: Nitric oxide-induced apoptosis is mediated by Bax/Bcl-2 gene expression, transition of cytochrome c, and activation of caspase-3 in rat vascular smooth muscle cells. Clin Chim Acta. 341:83–91. 2004. View Article : Google Scholar : PubMed/NCBI
21	Lin F, Song A, Wu J, Jiang X, Long J, Chen J, Duan Y, Shi Y and Deng L: ADAM33 protein expression and the mechanics of airway smooth muscle cells are highly correlated in ovalbumin-sensitized rats. Mol Med Rep. 8:1209–1215. 2013. View Article : Google Scholar : PubMed/NCBI
22	Zhou J, Bai W, Liu Q, Cui J and Zhang W: Silencing of ADAM33 restrains proliferation and induces apoptosis of airway smooth muscle cells in ovalbumin-induced asthma model. J Cell Biochem. Nov 18–2018.(Epub ahead of print).
23	Livak KJ and Schmittgen TD: Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) method. Methods. 25:402–408. 2001. View Article : Google Scholar : PubMed/NCBI
24	Tamhane AC: Statistical analysis of designed experiments: Theory and applications. Higher Education Press; Beijing: 2006
25	Dijkstra A, Postma DS, Noordhoek JA, Lodewijk ME, Kauffman HF, ten Hacken NH and Timens W: Expression of ADAMs (‘a disintegrin and metalloprotease’) in the human lung. Virchows Arch. 454:441–449. 2009. View Article : Google Scholar : PubMed/NCBI
26	Foley SC, Mogas AK, Olivenstein R, Fiset PO, Chakir J, Bourbeau J, Ernst P, Lemière C, Martin JG and Hamid Q: Increased expression of ADAM33 and ADAM8 with disease progression in asthma. J Allergy Clin Immunol. 119:863–871. 2007. View Article : Google Scholar : PubMed/NCBI
27	Van Eerdewegh P, Little RD, Dupuis J, Del Mastro RG, Falls K, Simon J, Torrey D, Pandit S, McKenny J, Braunschweiger K, et al: Association of the ADAM33 gene with asthma and bronchial hyperresponsiveness. Nature. 418:426–430. 2002. View Article : Google Scholar : PubMed/NCBI
28	Wen J, Muyesha P, Gong X, Hu X, Hao Y, Yan F, Zang L and Wang J: Effect of γ-interferon on the expression of ADAM33 gene in human airway wall vascular smooth muscle cells. Journal of Xinjiang Medical University. 42:965–970. 2019.
29	Tripathi P, Awasthi S and Gao P: ADAM metallopeptidase domain 33 (ADAM33): A promising target for asthma. Mediators Inflamm. 2014:5720252014. View Article : Google Scholar : PubMed/NCBI
30	Fang L, Wu J, Huang T, Zhang P, Xin X and Shi Y: TGF-β1 stimulates epithelial-mesenchymal transition mediated by ADAM33. Exp Ther Med. 15:985–992. 2018.PubMed/NCBI
31	Yang Y, Wicks J, Haitchi HM, Powell RM, Manuyakorn W, Howarth PH, Holgate ST and Davies DE: Regulation of a disintegrin and metalloprotease-33 expression by transforming growth factor-β. Am J Respir Cell Mol Biol. 46:633–640. 2012. View Article : Google Scholar : PubMed/NCBI
32	Pei QM, Jiang P, Yang M, Qian XJ, Liu JB, Zheng H, Zhao LH and Kim SH: Upregulation of a disintegrin and metalloproteinase-33 by VEGF in human airway smooth muscle cells: Implications for asthma. Cell Cycle. 15:2819–2826. 2016. View Article : Google Scholar : PubMed/NCBI
33	Kim SH, Pei QM, Jiang P, Yang M, Qian XJ and Liu JB: Effect of active vitamin D3 on VEGF-induced ADAM33 expression and proliferation in human airway smooth muscle cells: Implications for asthma treatment. Respir Res. 18:72017. View Article : Google Scholar : PubMed/NCBI
34	Duan Y, Long J, Chen J, Jiang X, Zhu J, Jin Y, Lin F, Zhong J, Xu R, Mao L and Deng L: Overexpression of soluble ADAM33 promotes a hypercontractile phenotype of the airway smooth muscle cell in rat. Exp Cell Res. 349:109–118. 2016. View Article : Google Scholar : PubMed/NCBI
35	Li W, Liang R, Huang H, Wu B and Zhong Y: Effects of IFN-gamma on cell growth and the expression of ADAM33 gene in human embryonic lung Mrc-5 fibroblasts in vitro. J Asthma. 55:15–25. 2018. View Article : Google Scholar : PubMed/NCBI
36	Yu JS and Cui W: Proliferation, survival and metabolism: The role of PI3K/AKT/mTOR signalling in pluripotency and cell fate determination. Development. 143:3050–3060. 2016. View Article : Google Scholar : PubMed/NCBI
37	Singh S, Bodas M, Bhatraju NK, Pattnaik B, Gheware A, Parameswaran PK, Thompson M, Freeman M, Mabalirajan U, Gosens R, et al: Hyperinsulinemia adversely affects lung structure and function. Am J Physiol Lung Cell Mol Physiol. 310:L837–L845. 2016. View Article : Google Scholar : PubMed/NCBI
38	Su X, Ren Y, Yu N, Kong L and Kang J: Thymoquinone inhibits inflammation, neoangiogenesis and vascular remodeling in asthma mice. Int Immunopharmacol. 38:70–80. 2016. View Article : Google Scholar : PubMed/NCBI
39	Edlich F: BCL-2 proteins and apoptosis: Recent insights and unknowns. Biochem Biophys Res Commun. 500:26–34. 2018. View Article : Google Scholar : PubMed/NCBI
40	Liu WJ, Zhu SY, Chen YL, Wu X, Ni WJ, Chen YF and Zhao L: The effects of leptin on apoptosis of airway smooth muscle cells via the PI3K/Akt signaling pathway. Zhonghua Jie He He Hu Xi Za Zhi. 35:915–918. 2012.(In Chinese). PubMed/NCBI
41	Wang S, Cheng Z and Chen X: Promotion of PTEN on apoptosis through PI3K/Akt signal in vascular smooth muscle cells of mice model of coronary heart disease. J Cell Biochem. 120:14636–14644. 2019. View Article : Google Scholar : PubMed/NCBI