Exosomes Derived from Human Adipose Mesenchymal Stem Cells Inhibits Fibrosis and Treats Oral Submucous Fibrosis via the miR-181a-5p/Smad2 Axis

被引:5
作者
Shao, Zifei [1 ,2 ]
Xu, Jinhao [1 ,2 ]
Xu, Xiaoyang [1 ,2 ]
Wang, Xiang [1 ,2 ]
Zhou, Yuxi [1 ,2 ]
Li, Yiyang [1 ,2 ]
Li, Kun [1 ,2 ,3 ]
机构
[1] Cent South Univ, Dept Oral & Maxillofacial Surg, Xiangya Stomatol Hosp, Changsha 410000, Peoples R China
[2] Cent South Univ, Sch Stomatol, Changsha 410000, Peoples R China
[3] Hunan Clin Res Ctr Oral Major Dis & Oral Hlth, Changsha, Peoples R China
关键词
ADSC-Exo; OSF; Myofibroblasts; miR-181a-5p; TGF-beta pathway; TGF-BETA; FIBROBLASTS; MECHANISMS; EXPRESSION; MICRORNAS; IMPACT; NUT;
D O I
10.1007/s13770-023-00579-0
中图分类号
Q813 [细胞工程];
学科分类号
摘要
BACKGROUND: Oral submucous fibrosis (OSF) is a chronic disease with carcinogenic tendency that poses a nonnegligible threat to human health. Exosomes derived from human adipose mesenchymal stem cells (ADSC-Exo) reduces visceral and cutaneous fibroses, but their role in OSF has received little attention. The aim of this study was to investigate the effects of ADSC-Exo on OSF and elucidate the mechanism. METHODS: In brief, ADSCs were extracted from adipose tissues and subjected to flow cytometry and induction culture. Fibroblasts were isolated from human buccal mucosa and subjected to immunofluorescence. Myofibroblasts were obtained from fibroblasts induced by arecoline and identified. Immunofluorescence assay confirmed that myofibroblasts could take up ADSC-Exo. The effects of ADSC-Exo on the proliferative and migratory capacities of myofibroblasts were examined using the Cell Counting Kit-8 and scratch assay. Real-time quantitative polymerase chain reaction (qPCR) was performed to evaluate mothers against decapentaplegic homolog 2 (Smad2), Smad3, Smad7, collagen type 1 (Col1), Col3, alpha smooth muscle actin (alpha-SMA), fibronectin, and vimentin. Western blotting was performed to detect phospho (p)- Smad2, Smad2, p-Smad2/3, Smad2/3, Smad7, Col1, Col3, a-SMA, fibronectin, and vimentin. Furthermore, the dual-luciferase reporter assay was performed to prove that miR-181a-5p in ADSC-Exo directly inhibited the expression of Smad2 mRNA to regulate the transforming growth factor beta (TGF-beta) pathway. We also performed qPCR and western blotting to verify the results. RESULTS: ADSC-Exo could promote the proliferation and migration of myofibroblasts, reduce the expressions of p-smad2, Smad2, p-smad2/3, Smad2/3, Col1, aSMA, fibronectin, and vimentin and elevated the levels of Smad7 and Col3. In addition, miR-181a-5p was highly expressed in ADSC-Exo and bound to the 3'-untranslated region of Smad2. ADSCExo enriched with miR-181a-5p reduced collagen production in myofibroblasts and modulated the TGF-b pathway. CONCLUSIONS: ADSC-Exo promoted the proliferative and migratory capacities of myofibroblasts and inhibited collagen deposition and trans-differentiation of myofibroblasts in vitro. miR-181a-5p in exosomes targets Smad2 to regulate the TGF-b pathway in myofibroblasts. ADSC-Exo perform antifibrotic actions through the miR-181a-5p/Smad2 axis and may be a promising clinical treatment for OSF.
引用
收藏
页码:123 / 135
页数:13
相关论文
共 56 条
[1]   The Role of Connective Tissue Growth Factor (CTGF/CCN2) in Skeletogenesis [J].
Arnott, John A. ;
Lambi, Alex G. ;
Mundy, Christina ;
Hendesi, Honey ;
Pixley, Robin A. ;
Owen, Thomas A. ;
Safadi, Fayez F. ;
Popoff, Steven N. .
CRITICAL REVIEWS IN EUKARYOTIC GENE EXPRESSION, 2011, 21 (01) :43-69
[2]   The impact of microRNAs on protein output [J].
Baek, Daehyun ;
Villen, Judit ;
Shin, Chanseok ;
Camargo, Fernando D. ;
Gygi, Steven P. ;
Bartel, David P. .
NATURE, 2008, 455 (7209) :64-U38
[3]   TGF-β1-Containing Exosomes from Injured Epithelial Cells Activate Fibroblasts to Initiate Tissue Regenerative Responses and Fibrosis [J].
Borges, Fernanda T. ;
Melo, Sonia A. ;
Oezdemir, Berna C. ;
Kato, Noritoshi ;
Revuelta, Ignacio ;
Miller, Caroline A. ;
Gattone, Vincent H., II ;
LeBleu, Valerie S. ;
Kalluri, Raghu .
JOURNAL OF THE AMERICAN SOCIETY OF NEPHROLOGY, 2013, 24 (03) :385-392
[4]   ENDOTOXIN-INDUCED SERUM FACTOR THAT CAUSES NECROSIS OF TUMORS [J].
CARSWELL, EA ;
OLD, LJ ;
KASSEL, RL ;
GREEN, S ;
FIORE, N ;
WILLIAMSON, B .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1975, 72 (09) :3666-3670
[5]   Arecoline- induced myofibroblast transdifferentiation from human buccal mucosal fibroblasts is mediated by ZEB1 [J].
Chang, Yu-Chao ;
Tsai, Chung-Hung ;
Lai, You-Liang ;
Yu, Cheng-Chia ;
Chi, Wan-Yu ;
Li, Jung Jung ;
Chang, Wen-Wei .
JOURNAL OF CELLULAR AND MOLECULAR MEDICINE, 2014, 18 (04) :698-708
[6]   The Role of MicroRNA-181a in Myocardial Fibrosis Following Myocardial Infarction in a Rat Model [J].
Chen, Peng ;
Pan, Jialin ;
Zhang, Xinming ;
Shi, Zhewei ;
Yang, Xiangjun .
MEDICAL SCIENCE MONITOR, 2018, 24 :4121-4127
[7]  
Dai MJ, 2017, TISSUE ENG PT A, V23, P1221, DOI [10.1089/ten.tea.2017.0045, 10.1089/ten.TEA.2017.0045]
[8]   Exosomes from adipose-derived mesenchymal stem cells ameliorate cardiac damage after myocardial infarction by activating S1P/SK1/S1PR1 signaling and promoting macrophage M2 polarization [J].
Deng, Shengqiong ;
Zhou, Xianjin ;
Ge, Zhiru ;
Song, Yuting ;
Wang, Hairong ;
Liu, Xinghui ;
Zhang, Denghai .
INTERNATIONAL JOURNAL OF BIOCHEMISTRY & CELL BIOLOGY, 2019, 114
[9]   Autophagy Regulates TGF-β Expression and Suppresses Kidney Fibrosis Induced by Unilateral Ureteral Obstruction [J].
Ding, Yan ;
Kim, Sung Il ;
Lee, So-Young ;
Koo, Ja Kun ;
Wang, Zhibo ;
Choi, Mary E. .
JOURNAL OF THE AMERICAN SOCIETY OF NEPHROLOGY, 2014, 25 (12) :2835-2846
[10]   Extracellular vesicles, the liquid biopsy of the future [J].
Eguchi, Akiko ;
Kostallari, Enis ;
Feldstein, Ariel E. ;
Shah, Vijay H. .
JOURNAL OF HEPATOLOGY, 2019, 70 (06) :1292-1294