Paracrine factors from mesenchymal stem cells attenuate epithelial injury and lung fibrosis

Shen,Qinqin; Chen,Bing; Xiao,Zhifeng; Zhao,Lifen; Xu,Xuefeng; Wan,Xuan; Jin,Mulan; Dai,Jianwu; Dai,Huaping

doi:10.3892/mmr.2014.3092

Paracrine factors from mesenchymal stem cells attenuate epithelial injury and lung fibrosis

Authors:
- Qinqin Shen
- Bing Chen
- Zhifeng Xiao
- Lifen Zhao
- Xuefeng Xu
- Xuan Wan
- Mulan Jin
- Jianwu Dai
- Huaping Dai
View Affiliations

Affiliations: Beijing Key Laboratory of Respiratory and Pulmonary Circulation, Beijing Institute of Respiratory Medicine, Department of Respiratory and Critical Care Medicine, Beijing Chao‑Yang Hospital, Capital Medical University, Beijing 100020, P.R. China, Key Laboratory of Molecular Development Biology, Institute of Genetics and Developmental Biology, Chinese Academy of Sciences, Beijing 100190, P.R. China, Department of Pathology Medicine, Beijing Chao‑Yang Hospital, Capital Medical University, Beijing 100020, P.R. China
Published online on: December 15, 2014 https://doi.org/10.3892/mmr.2014.3092
Pages: 2831-2837

Metrics: Total Views: 0 (Spandidos Publications: | PMC Statistics: )

Metrics: Total PDF Downloads: 0 (Spandidos Publications: | PMC Statistics: )

Cited By (CrossRef): 0 citations Loading Articles...

Abstract

Paracrine factors are currently considered to be the major mechanism through which mesenchymal stem cells (MSCs) exert their actions. The aim of this study was to investigate the protective effects of conditioned medium (CM) from bone marrow mesenchymal stem cells (MSC) on bleomycin (BLM)‑induced lung injury and fibrosis, both in vitro and in vivo. A549 human non‑small cell lung cancer epithelial cells were cultured in serum‑free medium, or MSC‑CM, both with or without BLM. The protective effects of MSC‑CM was determined by MTT assay to assess cell viability and Annexin V‑PE to assess apoptosis. Rats were intratracheally injected with MSC‑CM, saline, or conditioned medium from fibroblasts on day 0 and day 3 after intratracheal administration of BLM, and were sacrificed on day 28. Lung injury and fibrosis were assessed by histological assessment, Ashcroft score, and hydroxyproline assay; lung cell apoptosis was detected using terminal deoxynucleotidyl transferase dUTP nick end labeling assay. In comparison to the control group (0.17±0.01), 8 and 16% MSC‑CM had a significant stimulatory effect on A549 cellular proliferation (0.24±0.03 and 0.24±0.04, respectively, P<0.01). A549 cells cultured with MSC‑CM were protected from BLM‑induced apoptosis, 23.43±3.76% vs. 38.06±4.32%; (P<0.05). In the BLM‑challenged rats, MSC‑CM was shown to protect against lung fibrosis in terms of lung inflammation, fibrotic scores, collagen deposition, and cell apoptosis. This data suggests that MSCs are capable of protecting against lung injury and fibrosis both in vitro and in vivo through a paracrine anti‑inflammatory mechanism. MSC‑CM may provide a novel approach for the treatment of lung fibrosis.

View Figures

View References

1	King TE Jr, Pardo A and Selman M: Idiopathic pulmonary fibrosis. Lancet. 378:1949–1961. 2011. View Article : Google Scholar : PubMed/NCBI
2	Raghu G, Collard HR, Egan JJ, et al; ATS/ERS/JRS/ALAT Committee on Idiopathic Pulmonary Fibrosis. An official ATS/ERS/JRS/ALAT statement: idiopathic pulmonary fibrosis: evidence-based guidelines for diagnosis and management. Am J Respir Crit Care Med. 183:788–824. 2011. View Article : Google Scholar : PubMed/NCBI
3	Kropski JA, Lawson WE, Young LR and Blackwell TS: Genetic studies provide clues on the pathogenesis of idiopathic pulmonary fibrosis. Dis Model Mech. 6:9–17. 2013. View Article : Google Scholar :
4	Zoz DF, Lawson WE and Blackwell TS: Idiopathic pulmonary fibrosis: a disorder of epithelial cell dysfunction. Am J Med Sci. 341:435–438. 2011. View Article : Google Scholar : PubMed/NCBI
5	Hardie WD, Hagood JS, Dave V, et al: Signaling pathways in the epithelial origins of pulmonary fibrosis. Cell Cycle. 9:2769–2776. 2010. View Article : Google Scholar : PubMed/NCBI
6	Sakai N and Tager AM: Fibrosis of two: Epithelial cell-fibroblast interactions in pulmonary fibrosis. Biochim Biophys Acta. 1832:911–921. 2013. View Article : Google Scholar : PubMed/NCBI
7	Maher TM, Evans IC, Bottoms SE, et al: Diminished prostaglandin E2 contributes to the apoptosis paradox in idiopathic pulmonary fibrosis. Am J Respir Crit Care Med. 182:73–82. 2010. View Article : Google Scholar : PubMed/NCBI
8	Li X, Shu R, Filippatos G and Uhal BD: Apoptosis in lung injury and remodeling. J Appl Physiol (1985). 97:1535–1542. 2004. View Article : Google Scholar
9	Degryse AL, Tanjore H, Xu XC, et al: Repetitive intratracheal bleomycin models several features of idiopathic pulmonary fibrosis. Am J Physiol Lung Cell Mol Physiol. 299:L442–452. 2010. View Article : Google Scholar : PubMed/NCBI
10	Serrano-Mollar A, Nacher M, Gay-Jordi G, et al: Intratracheal transplantation of alveolar type II cells reverses bleomycin-induced lung fibrosis. Am J Respir Crit Care Med. 176:1261–1268. 2007. View Article : Google Scholar : PubMed/NCBI
11	Bao S, Wang Y, Sweeney P, et al: Keratinocyte growth factor induces Akt kinase activity and inhibits Fas-mediated apoptosis in A549 lung epithelial cells. Am J Physiol Lung Cell Mol Physiol. 288:L36–L42. 2005. View Article : Google Scholar
12	Crestani B, Marchand-Adam S, Quesnel C, et al: Hepatocyte growth factor and lung fibrosis. Proc Am Thorac Soc. 9:158–163. 2012. View Article : Google Scholar : PubMed/NCBI
13	Umeda Y, Marui T, Matsuno Y, et al: Skeletal muscle targeting in vivo electroporation-mediated HGF gene therapy of bleomycin-induced pulmonary fibrosis in mice. Lab Invest. 84:836–844. 2004. View Article : Google Scholar : PubMed/NCBI
14	Krause DS, Theise ND, Collector MI, et al: Multi-organ, multi-lineage engraftment by a single bone marrow-derived stem cell. Cell. 105:369–377. 2001. View Article : Google Scholar : PubMed/NCBI
15	Rejman J, Colombo C and Conese M: Engraftment of bone marrow-derived stem cells to the lung in a model of acute respiratory infection by Pseudomonas aeruginosa. Mol Ther. 17:1257–1265. 2009. View Article : Google Scholar : PubMed/NCBI
16	Fritzell JA Jr, Mao Q, Gundavarapu S, et al: Fate and effects of adult bone marrow cells in lungs of normoxic and hyperoxic newborn mice. Am J Respir Cell Mol Biol. 40:575–587. 2009. View Article : Google Scholar :
17	Tzouvelekis A, Ntolios P and Bouros D: Stem cell treatment for chronic lung diseases. Respiration. 85:179–192. 2013. View Article : Google Scholar : PubMed/NCBI
18	Conese M, Carbone A, Castellani S and Di Gioia S: Paracrine effects and heterogeneity of marrow-derived stem/progenitor cells: relevance for the treatment of respiratory diseases. Cells Tissues Organs. 197:445–473. 2013. View Article : Google Scholar : PubMed/NCBI
19	van Poll D, Parekkadan B, Cho CH, et al: Mesenchymal stem cell-derived molecules directly modulate hepatocellular death and regeneration in vitro and in vivo. Hepatology. 47:1634–1643. 2008. View Article : Google Scholar : PubMed/NCBI
20	Xu J, Woods CR, Mora AL, et al: Prevention of endotoxin-induced systemic response by bone marrow-derived mesenchymal stem cells in mice. Am J Physiol Lung Cell Mol Physiol. 293:L131–L141. 2007. View Article : Google Scholar : PubMed/NCBI
21	van Haaften T, Byrne R, Bonnet S, et al: Airway delivery of mesenchymal stem cells prevents arrested alveolar growth in neonatal lung injury in rats. Am J Respir Crit Care Med. 180:1131–1142. 2009. View Article : Google Scholar : PubMed/NCBI
22	Tögel F, Weiss K, Yang Y, et al: Vasculotropic, paracrine actions of infused mesenchymal stem cells are important to the recovery from acute kidney injury. Am J Physiol Renal Physiol. 292:F1626–F1635. 2007. View Article : Google Scholar : PubMed/NCBI
23	Ashcroft T, Simpson JM and Timbrell V: Simple method of estimating severity of pulmonary fibrosis on a numerical scale. J Clin Pathol. 41:467–470. 1988. View Article : Google Scholar : PubMed/NCBI
24	Phinney DG: Building a consensus regarding the nature and origin of mesenchymal stem cells. J Cell Biochem Suppl. 38:7–12. 2002. View Article : Google Scholar : PubMed/NCBI
25	Nauta AJ and Fibbe WE: Immunomodulatory properties of mesenchymal stromal cells. Blood. 110:3499–3506. 2007. View Article : Google Scholar : PubMed/NCBI
26	Lau AN, Goodwin M, Kim CF and Weiss DJ: Stem cells and regenerative medicine in lung biology and diseases. Mol Ther. 20:1116–1130. 2012. View Article : Google Scholar : PubMed/NCBI
27	Sadat S, Gehmert S, Song YH, et al: The cardioprotective effect of mesenchymal stem cells is mediated by IGF-I and VEGF. Biochem Biophys Res Commun. 363:674–679. 2007. View Article : Google Scholar : PubMed/NCBI
28	Nakanishi C, Yamagishi M, Yamahara K, et al: Activation of cardiac progenitor cells through paracrine effects of mesenchymal stem cells. Biochem Biophys Res Commun. 374:11–16. 2008. View Article : Google Scholar : PubMed/NCBI
29	Xu YX, Chen L, Wang R, et al: Mesenchymal stem cell therapy for diabetes through paracrine mechanisms. Med Hypotheses. 71:390–393. 2008. View Article : Google Scholar : PubMed/NCBI
30	Gazdhar A, Susuri N, Hostettler K, et al: HGF expressing stem cells in usual interstitial pneumonia originate from the bone marrow and are antifibrotic. PLoS One. 8:e654532013. View Article : Google Scholar : PubMed/NCBI
31	Gazdhar A, Temuri A, Knudsen L, et al: Targeted gene transfer of hepatocyte growth factor to alveolar type II epithelial cells reduces lung fibrosis in rats. Hum Gene Ther. 24:105–116. 2013. View Article : Google Scholar
32	Chakraborty S, Chopra P, Hak A, Dastidar SG and Ray A: Hepatocyte growth factor is an attractive target for the treatment of pulmonary fibrosis. Expert Opin Investig Drugs. 22:499–515. 2013. View Article : Google Scholar : PubMed/NCBI
33	Sakamoto S, Yazawa T, Baba Y, et al: Keratinocyte growth factor gene transduction ameliorates pulmonary fibrosis induced by bleomycin in mice. Am J Respir Cell Mol Biol. 45:489–497. 2011. View Article : Google Scholar
34	Inghilleri S, Morbini P, Oggionni T, Barni S and Fenoglio C: In situ assessment of oxidant and nitrogenic stress in bleomycin pulmonary fibrosis. Histochem Cell Biol. 125:661–669. 2006. View Article : Google Scholar
35	Teixeira KC, Soares FS, Rocha LG, et al: Attenuation of bleomycin-induced lung injury and oxidative stress by N-acetylcysteine plus deferoxamine. Pulm Pharmacol Ther. 21:309–316. 2008. View Article : Google Scholar
36	Lanza C, Morando S, Voci A, et al: Neuroprotective mesenchymal stem cells are endowed with a potent antioxidant effect in vivo. J Neurochem. 110:1674–1684. 2009. View Article : Google Scholar : PubMed/NCBI
37	Ortiz LA, Gambelli F, McBride C, et al: Mesenchymal stem cell engraftment in lung is enhanced in response to bleomycin exposure and ameliorates its fibrotic effects. Proc Natl Acad Sci USA. 100:8407–8411. 2003. View Article : Google Scholar : PubMed/NCBI
38	Kumamoto M, Nishiwaki T, Matsuo N, Kimura H and Matsushima K: Minimally cultured bone marrow mesenchymal stem cells ameliorate fibrotic lung injury. Eur Respir J. 34:740–748. 2009. View Article : Google Scholar : PubMed/NCBI
39	Lee SH, Jang AS, Kim YE, et al: Modulation of cytokine and nitric oxide by mesenchymal stem cell transfer in lung injury/fibrosis. Respir Res. 11:162010. View Article : Google Scholar : PubMed/NCBI
40	Moodley Y, Atienza D, Manuelpillai U, et al: Human umbilical cord mesenchymal stem cells reduce fibrosis of bleomycin-induced lung injury. Am J Pathol. 175:303–313. 2009. View Article : Google Scholar : PubMed/NCBI