Adipose mesenchymal stem cells-derived exosomes attenuated hyperoxia-induced lung injury in neonatal rats via inhibiting the NF-κB signaling pathway
Cuie Chen MD
Department of Pediatrics, Yiwu Maternity and Children Hospital, Jinhua, Zhejiang, China
Contribution: Conceptualization, Methodology, Investigation, Funding acquisition, Writing - original draft
Search for more papers by this authorYuxia Jin MD
Department of Prenatal Diagnostic Center, Yiwu Maternity and Children Hospital, Jinhua, Zhejiang, China
Contribution: Investigation, Methodology, Validation, Visualization, Funding acquisition
Search for more papers by this authorHongxing Jin BS
Department of Pediatrics, Yiwu Maternity and Children Hospital, Jinhua, Zhejiang, China
Contribution: Investigation, Methodology, Validation
Search for more papers by this authorShujun Chen BS
Department of Pediatrics, Yiwu Maternity and Children Hospital, Jinhua, Zhejiang, China
Contribution: Methodology, Validation, Visualization, Software
Search for more papers by this authorLu Wang BS
Department of Prenatal Diagnostic Center, Yiwu Maternity and Children Hospital, Jinhua, Zhejiang, China
Contribution: Validation, Visualization, Software, Formal analysis
Search for more papers by this authorLiuqing Ji BS
Department of Pediatrics, Yiwu Maternity and Children Hospital, Jinhua, Zhejiang, China
Contribution: Validation, Visualization, Software, Formal analysis
Search for more papers by this authorShi Wang MD
Department of Anesthesiology, Women's Hospital School of Medicine Zhejiang University, Hangzhou, Zhejiang, China
Contribution: Project administration, Data curation, Supervision, Resources
Search for more papers by this authorXixi Zhang BS
Department of Pediatrics, Yuhuan People's Hospital, Taizhou, Zhejiang, China
Contribution: Data curation, Supervision, Resources
Search for more papers by this authorAnqun Sheng MD
Department of Pediatrics, The First Affiliated Hospital of Wenzhou Medical University, Wenzhou, Zhejiang, China
Contribution: Data curation, Software, Formal analysis
Search for more papers by this authorCorresponding Author
Yuanyuan Sun PhD
Department of Pediatrics, The First Affiliated Hospital of Wenzhou Medical University, Wenzhou, Zhejiang, China
Department of Pediatrics, The Quzhou Affiliated Hospital of Wenzhou Medical University (Quzhou People's Hospital), Quzhou, Zhejiang, China
Correspondence Yuanyuan Sun, Department of Pediatrics, The Quzhou Affiliated Hospital of Wenzhou Medical University (Quzhou People's Hospital), No. 100, Minjiang Ave, Kecheng Smart New City, Quzhou, Zhejiang 324000, China.
Email: [email protected]
Contribution: Conceptualization, Investigation, Writing - original draft, Funding acquisition, Writing - review & editing
Search for more papers by this authorCuie Chen MD
Department of Pediatrics, Yiwu Maternity and Children Hospital, Jinhua, Zhejiang, China
Contribution: Conceptualization, Methodology, Investigation, Funding acquisition, Writing - original draft
Search for more papers by this authorYuxia Jin MD
Department of Prenatal Diagnostic Center, Yiwu Maternity and Children Hospital, Jinhua, Zhejiang, China
Contribution: Investigation, Methodology, Validation, Visualization, Funding acquisition
Search for more papers by this authorHongxing Jin BS
Department of Pediatrics, Yiwu Maternity and Children Hospital, Jinhua, Zhejiang, China
Contribution: Investigation, Methodology, Validation
Search for more papers by this authorShujun Chen BS
Department of Pediatrics, Yiwu Maternity and Children Hospital, Jinhua, Zhejiang, China
Contribution: Methodology, Validation, Visualization, Software
Search for more papers by this authorLu Wang BS
Department of Prenatal Diagnostic Center, Yiwu Maternity and Children Hospital, Jinhua, Zhejiang, China
Contribution: Validation, Visualization, Software, Formal analysis
Search for more papers by this authorLiuqing Ji BS
Department of Pediatrics, Yiwu Maternity and Children Hospital, Jinhua, Zhejiang, China
Contribution: Validation, Visualization, Software, Formal analysis
Search for more papers by this authorShi Wang MD
Department of Anesthesiology, Women's Hospital School of Medicine Zhejiang University, Hangzhou, Zhejiang, China
Contribution: Project administration, Data curation, Supervision, Resources
Search for more papers by this authorXixi Zhang BS
Department of Pediatrics, Yuhuan People's Hospital, Taizhou, Zhejiang, China
Contribution: Data curation, Supervision, Resources
Search for more papers by this authorAnqun Sheng MD
Department of Pediatrics, The First Affiliated Hospital of Wenzhou Medical University, Wenzhou, Zhejiang, China
Contribution: Data curation, Software, Formal analysis
Search for more papers by this authorCorresponding Author
Yuanyuan Sun PhD
Department of Pediatrics, The First Affiliated Hospital of Wenzhou Medical University, Wenzhou, Zhejiang, China
Department of Pediatrics, The Quzhou Affiliated Hospital of Wenzhou Medical University (Quzhou People's Hospital), Quzhou, Zhejiang, China
Correspondence Yuanyuan Sun, Department of Pediatrics, The Quzhou Affiliated Hospital of Wenzhou Medical University (Quzhou People's Hospital), No. 100, Minjiang Ave, Kecheng Smart New City, Quzhou, Zhejiang 324000, China.
Email: [email protected]
Contribution: Conceptualization, Investigation, Writing - original draft, Funding acquisition, Writing - review & editing
Search for more papers by this authorCuie Chen and Yuxia Jin contributed equally to this study and are co-first authors.
Abstract
Objective
Bronchopulmonary dysplasia (BPD) is the most common chronic morbidity in extremely preterm infants. Mesenchymal stem cells-derived exosomes (MSC-Exos) therapies have shown prospects in animal models of BPD. Our study aimed to evaluate the effect of adipose mesenchymal stem cells-derived exosomes (AMSC-Exos) on BPD and the role of the NF-κB signaling pathway in this process.
Methods
The AMSCs were extracted and AMSC-Exos were isolated by ultracentrifugation method. Newborn rats were exposed to hyperoxia (90% O2) continuously for 7 days to establish a BPD model. The rats were treated with AMSC-Exos by intratracheal administration on postnatal day 4 (P4). Pulmonary morphology, pulmonary vasculature, inflammatory factors, and NF-κB were assessed. Hyperoxia-induced primary type II alveolar epithelial cells (AECIIs) and AMSC-Exos treatment with or without a pan-NF-κB inhibitor (PDTC) were established to explore the potential mechanism.
Results
Hyperoxia-exposed rats showed alveolar simplification with decreased radial alveolar count and increased mean linear intercept, low CD31, and vascular endothelial growth factor expression, reduced microvessel density, increased the expression of TNF-α, IL-1β, and IL-6 and decreased the expression of IL-10, and induced NF-κB phosphorylation. AMSC-Exos protected the neonatal lung from the hyperoxia-induced arrest of alveolar and vascular development, alleviated inflammation, and inhibited NF-κB phosphorylation. Hyperoxia decreased viability, increased apoptosis, enhanced inflammation, and induced NF-κB phosphorylation of AECIIs but improved by AMSC-Exos, PDTC, or AMSC-Exos+PDTC. The effect of AMSC-Exos+PDTC in AECIIs was the same as AMSC-Exos, but more notable than PDTC alone.
Conclusion
AMSC-Exos attenuated the hyperoxia-induced lung injury in neonatal rats by inhibiting the NF-κB signaling pathway partly.
CONFLICT OF INTEREST STATEMENT
The authors declare no conflict of interest.
Open Research
DATA AVAILABILITY STATEMENT
The data that supports the findings of this study are available in the supplementary material of this article.
Supporting Information
| Filename | Description |
|---|---|
| ppul27057-sup-0001-Figure_S1.jpg32.1 KB | Figure S1. The flow chart of AMSC-Exos treatment on type II alveolar epithelial cells (AECIIs) under hyperoxia. (Hyp: Hyperoxia). |
| ppul27057-sup-0002-Figure_S2.jpg431.5 KB | Figure S2. The characterizations of AMSCs. A-B: The morphology of AMSCs was uniform and long fusiform under optical microscope. A (100X), B (200X). C: The immunofluorescence showed that the AMSCs were strongly positive for CD90 with positive rate > 90% (200X). D: The differentiation ability of adipogenesis of AMSCs was good based on oil red O staining. E: The differentiation ability of osteogenesis of AMSCs was good based on Alizarin red staining. F: The differentiation ability of chondrogenesis of AMSCs was good based on Alcian blue staining. |
| ppul27057-sup-0003-Figure_S3.jpg64.6 KB | Figure S3. The characterizations of AECIIs. A. The morphology of AECIIs under optical microscope (100X). B. The immunofluorescence showed that the AECIIs were strongly positive for specific biomarkers SP-C with positive rate > 90% (200X). |
| ppul27057-sup-0004-Figure_S4.jpg222.7 KB | Figure S4. The effects of PDTC on cell proliferation and apoptosis of hyperoxia-induced AECIIs. A. The cell viability was determined by CCK-8 assays. B-C. The cell apoptosis rate was detected by flow cytometry. *P<0.05, ** P<0.01. 5 replicates. |
Please note: The publisher is not responsible for the content or functionality of any supporting information supplied by the authors. Any queries (other than missing content) should be directed to the corresponding author for the article.
REFERENCES
- 1Bell EF, Hintz SR, Hansen NI, et al. Mortality, in-hospital morbidity, care practices, and 2-year outcomes for extremely preterm infants in the US, 2013-2018. JAMA. 2022; 327(3): 248-263. doi:10.1001/jama.2021.23580
- 2Cao Y, Jiang S, Sun J, et al. Assessment of neonatal intensive care unit practices, morbidity, and mortality among very preterm infants in China. JAMA Network Open. 2021; 4(8):e2118904. doi:10.1001/jamanetworkopen.2021.18904
- 3Dankhara N, Holla I, Ramarao S, Kalikkot Thekkeveedu R. Bronchopulmonary dysplasia: pathogenesis and pathophysiology. J Clin Med. 2023; 12(13):4207. doi:10.3390/jcm12134207
- 4Jensen EA, Laughon MM, DeMauro SB, et al. Contributions of the NICHD neonatal research network to the diagnosis, prevention, and treatment of bronchopulmonary dysplasia. Semin Perinatol. 2022; 46(7):151638. doi:10.1016/j.semperi.2022.151638
- 5Omar SA, Abdul-Hafez A, Ibrahim S, et al. Stem-cell therapy for bronchopulmonary dysplasia (BPD) in newborns. Cells. 2022; 11(8):1275. doi:10.3390/cells11081275
- 6Ahn SY, Chang YS, Kim JH, Sung SI, Park WS. Two-year follow-up outcomes of premature infants enrolled in the phase I trial of mesenchymal stem cells transplantation for bronchopulmonary dysplasia. J Pediatr. 2017; 185: 49-54.e2. doi:10.1016/j.jpeds.2017.02.061
- 7Ahn SY, Chang YS, Lee MH, et al. Stem cells for bronchopulmonary dysplasia in preterm infants: a randomized controlled phase II trial. Stem Cells Transl Med. 2021; 10(8): 1129-1137. doi:10.1002/sctm.20-0330
- 8Lotfy A, AboQuella NM, Wang H. Mesenchymal stromal/stem cell (MSC)-derived exosomes in clinical trials. Stem Cell Res Ther. 2023; 14(1): 66. doi:10.1186/s13287-023-03287-7
- 9Mendt M, Rezvani K, Shpall E. Mesenchymal stem cell-derived exosomes for clinical use. Bone Marrow Transplant. 2019; 54(suppl 2): 789-792. doi:10.1038/s41409-019-0616-z
- 10Porzionato A, Zaramella P, Dedja A, et al. Intratracheal administration of clinical-grade mesenchymal stem cell-derived extracellular vesicles reduces lung injury in a rat model of bronchopulmonary dysplasia. Am J Physiol-Lung Cell Mol Physiol. 2019; 316(1): L6-L19. doi:10.1152/ajplung.00109.2018
- 11Sharma M, Bellio MA, Benny M, et al. Mesenchymal stem cell-derived extracellular vesicles prevent experimental bronchopulmonary dysplasia complicated by pulmonary hypertension. Stem Cells Transl Med. 2022; 11(8): 828-840. doi:10.1093/stcltm/szac041
- 12Bunnell BA. Adipose tissue-derived mesenchymal stem cells. Cells. 2021; 10(12):3433. doi:10.3390/cells10123433
- 13Trzyna A, Banaś-Ząbczyk A. Adipose-derived stem cells secretome and its potential application in “stem cell-free therapy”. Biomolecules. 2021; 11(6):878. doi:10.3390/biom11060878
- 14Li C, Li W, Pu G, Wu J, Qin F. Exosomes derived from miR-338-3p-modified adipose stem cells inhibited inflammation injury of chondrocytes via targeting RUNX2 in osteoarthritis. J Orthop Surg Res. 2022; 17(1): 567. doi:10.1186/s13018-022-03437-2
- 15Li R, Li D, Wang H, et al. Exosomes from adipose-derived stem cells regulate M1/M2 macrophage phenotypic polarization to promote bone healing via miR-451a/MIF. Stem Cell Res Ther. 2022; 13(1): 149. doi:10.1186/s13287-022-02823-1
- 16Yuan T, Meijia L, Xinyao C, Xinyue C, Lijun H. Exosome derived from human adipose-derived stem cell improve wound healing quality: a systematic review and meta-analysis of preclinical animal studies. Int Wound J. 2023; 20(6): 2424-2439. doi:10.1111/iwj.14081
- 17Savani RC. Modulators of inflammation in bronchopulmonary dysplasia. Semin Perinatol. 2018; 42(7): 459-470. doi:10.1053/j.semperi.2018.09.009
- 18Alvira CM. Nuclear factor-kappa-B signaling in lung development and disease: one pathway, numerous functions. Birth Defects Res Part A: Clin Mol Teratol. 2014; 100(3): 202-216. doi:10.1002/bdra.23233
- 19Mitchell S, Vargas J, Hoffmann A. Signaling via the NFκB system. WIREs Systems Biol Med. 2016; 8(3): 227-241. doi:10.1002/wsbm.1331
- 20Jiao B, Tang Y, Liu S, Guo C. Tetrandrine attenuates hyperoxia-induced lung injury in newborn rats via NF-κB p65 and ERK1/2 pathway inhibition. Ann Transl Med. 2020; 8(16): 1018. doi:10.21037/atm-20-5573
- 21Liu D, Wang Y, Li L, et al. Celecoxib protects hyperoxia-induced lung injury via NF-κB and AQP1. Front Pediatr. 2019; 7:228. doi:10.3389/fped.2019.00228
- 22Xue C, Gao Y, Li X, et al. Mesenchymal stem cells derived from adipose accelerate the progression of colon cancer by inducing a MT-CAFs phenotype via TRPC3/NF-KB axis. Stem Cell Res Ther. 2022; 13(1): 335. doi:10.1186/s13287-022-03017-5
- 23Liu J, Chen T, Lei P, Tang X, Huang P. Exosomes released by bone marrow mesenchymal stem cells attenuate lung injury induced by intestinal ischemia reperfusion via the TLR4/NF-κB pathway. Int J Med Sci. 2019; 16(9): 1238-1244. doi:10.7150/ijms.35369
- 24Sun Y, Chen C, Zhang X, et al. Heparin improves alveolarization and vascular development in hyperoxia-induced bronchopulmonary dysplasia by inhibiting neutrophil extracellular traps. Biochem Biophys Res Commun. 2020; 522(1): 33-39. doi:10.1016/j.bbrc.2019.11.041
- 25Braun RK, Chetty C, Balasubramaniam V, et al. Intraperitoneal injection of MSC-derived exosomes prevent experimental bronchopulmonary dysplasia. Biochem Biophys Res Commun. 2018; 503(4): 2653-2658. doi:10.1016/j.bbrc.2018.08.019
- 26Cooney TP, Thurlbeck WM. The radial alveolar count method of Emery and Mithal: a reappraisal 2: intrauterine and early postnatal lung growth. Thorax. 1982; 37(8): 580-583. doi:10.1136/thx.37.8.580
- 27Dunnill MS. Quantitative methods in the study of pulmonary pathology. Thorax. 1962; 17(4): 320-328. doi:10.1136/thx.17.4.320
- 28Hu Y, Xie L, Yu J, Fu H, Zhou D, Liu H. Inhibition of microRNA-29a alleviates hyperoxia-induced bronchopulmonary dysplasia in neonatal mice via upregulation of GAB1. Mol Med. 2019; 26(1): 3. doi:10.1186/s10020-019-0127-9
- 29Wang J, Zhang A, Huang F, Xu J, Zhao M. MSC-EXO and tempol ameliorate bronchopulmonary dysplasia in newborn rats by activating HIF-1α. Pediatr Pulmonol. 2023; 58(5): 1367-1379. doi:10.1002/ppul.26317
- 30Moreira A, Winter C, Joy J, et al. Intranasal delivery of human umbilical cord Wharton's jelly mesenchymal stromal cells restores lung alveolarization and vascularization in experimental bronchopulmonary dysplasia. Stem Cells Transl Med. 2020; 9(2): 221-234. doi:10.1002/sctm.18-0273
- 31Maniscalco WM, Watkins RH, Pryhuber GS, Bhatt A, Shea C, Huyck H. Angiogenic factors and alveolar vasculature: development and alterations by injury in very premature baboons. Am J Physiol-Lung Cell Mol Physiol. 2002; 282(4): L811-L823. doi:10.1152/ajplung.00325.2001
- 32Cui H, Huang W, He J. Expression of transcription factor CASZ1 and its relationship with pulmonary microvascular development in newborn rats after hyperoxia-exposure. Zhonghua er ke za zhi=Chinese J Pediatr. 2016; 54(1): 37-42. doi:10.3760/cma.j.issn.0578-1310.2016.01.009
- 33Zhang D, Zhao X, Zhang D, Gao S, Xue X, Fu J. Hyperoxia reduces STX17 expression and inhibits the autophagic flux in alveolar type II epithelial cells in newborn rats. Int J Mol Med. 2020; 46(2): 773-781. doi:10.3892/ijmm.2020.4617
- 34Fan HB, Zou Y, Han Q, et al. Cordyceps militaris immunomodulatory protein promotes the phagocytic ability of macrophages through the TLR4-NF-κB pathway. Int J Mol Sci. 2021; 22(22):12188. doi:10.3390/ijms222212188
- 35Anderson JD, Johansson HJ, Graham CS, et al. Comprehensive proteomic analysis of mesenchymal stem cell exosomes reveals modulation of angiogenesis via nuclear factor-kappaB signaling. Stem Cells. 2016; 34(3): 601-613. doi:10.1002/stem.2298
- 36Chen W, Huang Y, Han J, et al. Immunomodulatory effects of mesenchymal stromal cells-derived exosome. Immunol Res. 2016; 64(4): 831-840. doi:10.1007/s12026-016-8798-6
- 37Thébaud B, Goss KN, Laughon M, et al. Bronchopulmonary dysplasia. Nat Rev Dis Primers. 2019; 5(1): 78. doi:10.1038/s41572-019-0127-7
- 38Papagianis PC, Pillow JJ, Moss TJ. Bronchopulmonary dysplasia: pathophysiology and potential anti-inflammatory therapies. Paediatr Respir Rev. 2019; 30: 34-41. doi:10.1016/j.prrv.2018.07.007
- 39Hinz M, Arslan SÇ, Scheidereit C. It takes two to tango: IκBs, the multifunctional partners of NF-κB. Immunol Rev. 2012; 246(1): 59-76. doi:10.1111/j.1600-065X.2012.01102.x
- 40Wei H, Yin L, Feng S, et al. Dual-parallel inhibition of IL-10 and TGF-β1 controls LPS-induced inflammatory response via NF-κB signaling in grass carp monocytes/macrophages. Fish Shellfish Immunol. 2015; 44(2): 445-452. doi:10.1016/j.fsi.2015.03.023
- 41Sun J, Sun X, Chen J, et al. microRNA-27b shuttled by mesenchymal stem cell-derived exosomes prevents sepsis by targeting JMJD3 and downregulating NF-κB signaling pathway. Stem Cell Res Ther. 2021; 12(1): 14. doi:10.1186/s13287-020-02068-w
- 42Jiang Z, Zhang J. Mesenchymal stem cell-derived exosomes containing miR-145-5p reduce inflammation in spinal cord injury by regulating the TLR4/NF-κB signaling pathway. Cell Cycle. 2021; 20(10): 993-1009. doi:10.1080/15384101.2021.1919825
- 43Tao Y, Zhou J, Wang Z, et al. Human bone mesenchymal stem cells-derived exosomal miRNA-361-5p alleviates osteoarthritis by downregulating DDX20 and inactivating the NF-κB signaling pathway. Bioorg Chem. 2021; 113:104978. doi:10.1016/j.bioorg.2021.104978
- 44Wang X, Liu D, Zhang X, Yang L, Xia Z, Zhang Q. Exosomes from adipose-derived mesenchymal stem cells alleviate sepsis-induced lung injury in mice by inhibiting the secretion of IL-27 in macrophages. Cell Death Discov. 2022; 8(1): 18. doi:10.1038/s41420-021-00785-6
Citing Literature
