Mesenchymal stem cell-conditioned media suppresses inflammation-associated overproliferation of pulmonary artery smooth muscle cells in a rat model of pulmonary hypertension

Liu,Junfeng; Han,Zhibo; Han,Zhongchao; He,Zhixu

doi:10.3892/etm.2015.2953

Mesenchymal stem cell-conditioned media suppresses inflammation-associated overproliferation of pulmonary artery smooth muscle cells in a rat model of pulmonary hypertension

Authors:
- Junfeng Liu
- Zhibo Han
- Zhongchao Han
- Zhixu He
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Affiliations: Laboratory of Tissue Engineering and Stem Cells, Guiyang Medical College, Guiyang, Guizhou 550004, P.R. China, National Engineering Research Center of Cell Products, AmCellGene Co., Ltd., Tianjin 300457, P.R. China
Published online on: December 18, 2015 https://doi.org/10.3892/etm.2015.2953
Pages: 467-475
Copyright: © Liu et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

Inflammation-associated overproliferation of pulmonary artery smooth muscle cells (PASMCs) is considered to be involved in the pathogenesis of pulmonary hypertension (PH). The administration of mesenchymal stem cell‑conditioned media (MSC‑CM) has displayed benefits in the treatment of PH, however, the exact mechanism has yet to be elucidated. The present study aimed to determine whether MSC‑CM is able to suppress overproliferation of PASMCs in PH via immunoregulation. By the administration of MSC‑CM to monocrotaline (MCT)‑induced PH rats, and the development of an in vitro co‑culture system comprised of PASMCs and activated T cells, the therapeutic effects of MSC‑CM on PH, and the changes in the expression of correlated factors, including TNF‑α, calcineurin (CaN) and nuclear factor of activated T cells (NFAT), were assessed. Immunohistochemical staining results indicated that MSC‑CM was able to significantly suppress the production of TNF‑α in MCT‑induced PH and co‑culture systems; and reverse transcription‑quantitative polymerase chain reaction results showed significant downregulation of the expression of CaN and NFATc2 in PASMCs (P<0.01). Furthermore, MSC‑CM was able to significantly suppress CaN activity and NFATc2 activation (P<0.01), thus inhibiting the overproliferation of PASMCs. Finally, MSC‑CM improved abnormalities in hemodynamics and pulmonary histology in MCT‑induced PH. In conclusion, the findings of the current study suggest that administration of MSC‑CM has the potential to suppress inflammation‑associated overproliferation of PASMCs due to its immunosuppressive effects in PH and, thus, may serve as a beneficial therapeutic strategy.

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1	Bazan IS and Fares WH: Pulmonary hypertension: Diagnostic and therapeutic challenges. Ther Clin Risk Manag. 11:1221–1233. 2015.PubMed/NCBI
2	Tonelli AR: Pulmonary hypertension survival effects and treatment options in cystic fibrosis. Curr Opin Pulm Med. 19:652–661. 2013. View Article : Google Scholar : PubMed/NCBI
3	Tonelli AR, Fernandez-Bussy S, Lodhi S, Akindipe OA, Carrie RD, Hamilton K, Mubarak K and Baz MA: Prevalence of pulmonary hypertension in end-stage cystic fibrosis and correlation with survival. J Heart Lung Transplant. 29:865–872. 2010. View Article : Google Scholar : PubMed/NCBI
4	Hayes D Jr, Higgins RS, Kirkby S, McCoy KS, Wehr AM, Lehman AM and Whitson BA: Impact of pulmonary hypertension on survival in patients with cystic fibrosis undergoing lung transplantation, An analysis of the UNOS registry. J Cyst Fibros. 13:416–423. 2014. View Article : Google Scholar : PubMed/NCBI
5	Pinto RF: HiguchiM de L and Aiello VD: Decreased numbers of T-lymphocytes and predominance of recently recruited macrophages in the walls of peripheral pulmonary arteries from 26 patients with pulmonary hypertension secondary to congenital cardiac shunts. Cardiovasc Pathol. 13:268–275. 2004. View Article : Google Scholar : PubMed/NCBI
6	Perros F, Dorfmüller P, Montani D, Hammad H, Waelput W, Girerd B, Raymond N, Mercier O, Mussot S, Cohen-Kaminsky S, et al: Pulmonary lymphoid neogenesis in idiopathic pulmonary arterial hypertension. Am J Respir Crit Care Med. 185:311–321. 2012. View Article : Google Scholar : PubMed/NCBI
7	Heath D and Edwards JE: The pathology of hypertensive pulmonary vascular disease; a description of six grades of structural changes in the pulmonary arteries with special reference to congenital cardiac septal defects. Circulation. 18:533–547. 1958. View Article : Google Scholar : PubMed/NCBI
8	Soon E, Holmes AM, Treacy CM, Doughty NJ, Southgate L, Machado RD, Trembath RC, Jennings S, Barker L, Nicklin P, et al: Elevated levels of inflammatory cytokines predict survival in idiopathic and familial pulmonary arterial hypertension. Circulation. 122:920–927. 2010. View Article : Google Scholar : PubMed/NCBI
9	Frid MG, Brunetti JA, Burke DL, Carpenter TC, Davie NJ, Reeves JT and Roedersheimer MT: vanR ooijen N and Stenmark KR: Hypoxia-induced pulmonary vascular remodeling requires recruitment of circulating mesenchymal precursors of a monocyte/macrophage lineage. Am J Pathol. 168:659–669. 2006. View Article : Google Scholar : PubMed/NCBI
10	Rowlands DJ, Islam MN, Das SR, Huertas A, Quadri SK, Horiuchi K, Inamdar N, Emin MT, Lindert J, Ten VS, et al: Activation of TNFR1 ectodomain shedding by mitochondrial Ca²⁺ determines the severity of inflammation in mouse lung microvessels. J Clin Invest. 121:1986–1999. 2011. View Article : Google Scholar : PubMed/NCBI
11	Said SI, Hamidi SA and Gonzalez Bosc L: Asthma and pulmonary arterial, hypertension. Do they share a key mechanism of pathogenesis? Eur Respir J. 35:730–734. 2010. View Article : Google Scholar : PubMed/NCBI
12	Bonnet S, Rochefort G, Sutendra G, Archer SL, Haromy A, Webster L, Hashimoto K, Bonnet SN and Michelakis ED: The nuclear factor of activated T cells in pulmonary arterial hypertension can be therapeutically targeted. Proc Natl Acad Sci USA. 104:11418–11423. 2007. View Article : Google Scholar : PubMed/NCBI
13	de Frutos S, Spangler R, Alò D and Bosc LV: NFATc3 mediates chronic hypoxia-induced pulmonary arterial remodeling with alpha-actin up-regulation. J Biol Chem. 282:15081–15089. 2007. View Article : Google Scholar : PubMed/NCBI
14	Yang ZX, Han ZB, Ji YR, Wang YW, Liang L, Chi Y, Yang SG, Li LN, Luo WF, Li JP, et al: CD106 identifies a subpopulation of mesenchymal stem cells with unique immunomodulatory properties. PLoS One. 8:e593542013. View Article : Google Scholar : PubMed/NCBI
15	Aslam M, Baveja R, Liang OD, Fernandez-Gonzalez A, Lee C, Mitsialis SA and Kourembanas S: Bone marrow stromal cells attenuate lung injury in a murine model of neonatal chronic lung disease. Am J Respir Crit Care Med. 180:1122–1130. 2009. View Article : Google Scholar : PubMed/NCBI
16	Lu LL, Liu YJ, Yang SG, Zhao QJ, Wang X, Gong W, Han ZB, Xu ZS, Lu YX, Liu D, et al: Isolation and characterization of human umbilical cord mesenchymal stem cells with hematopoiesis-supportive function and other potentials. Haematologica. 91:1017–1026. 2006.PubMed/NCBI
17	Matsuda Y, Hoshikawa Y, Ameshima S, Suzuki S, Okada Y, Tabata T, Sugawara T, Matsumura Y and Kondo T: Effects of peroxisome proliferator-activated receptor gamma ligands on monocrotaline-induced pulmonary hypertension in rats. Nihon Kokyuki Gakkai Zasshi. 43:283–288. 2005.(In Japanese). PubMed/NCBI
18	Liu JF, DU ZD, Chen Z, Han ZC and He ZX: Granulocyte colony-stimulating factor attenuates monocrotaline-induced pulmonary hypertension by upregulating endothelial progenitor cells via the nitric oxide system. Exp Ther Med. 6:1402–1408. 2013.PubMed/NCBI
19	Maruyama H, Watanabe S, Kimura T, Liang J, Nagasawa T, Onodera M, Aonuma K and Yamaguchi I: Granulocyte colony-stimulating factor prevents progression of monocrotaline-induced pulmonary arterial hypertension in rats. Circ J. 71:138–143. 2007. View Article : Google Scholar : PubMed/NCBI
20	Woodrum DT, Ford JW, Cho BS, Hannawa KK, Stanley JC, Henke PK and Upchurch GR Jr: Differential effect of 17-beta-estradiol on smooth muscle cell and aortic explant MMP2. J Surg Res. 155:48–53. 2009. View Article : Google Scholar : PubMed/NCBI
21	Ahmad S, Choudhry MA, Shankar R and Sayeed MM: Transforming growth factor-beta negatively modulates T-cell responses in sepsis. FEBS Lett. 402:213–218. 1997. View Article : Google Scholar : PubMed/NCBI
22	Liu JF, Han ZB, Han ZC and He ZX: Mesenchymal stem cells suppress CaN/NFAT expression in the pulmonary arteries of rats with pulmonary hypertension. Exp Ther Med. 10:1657–1664. 2015.PubMed/NCBI
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	Tuder RM, Marecki JC, Richter A, Fijalkowska I and Flores S: Pathology of pulmonary hypertension. Clin Chest Med. 28:23–42. 2007. View Article : Google Scholar : PubMed/NCBI
25	Weiss DJ, Bertoncello I, Borok Z, Kim C, Panoskaltsis-Mortari A, Reynolds S, Rojas M, Stripp B, Warburton D and Prockop DJ: Stem cells and cell therapies in lung biology and lung diseases. Proc Am Thorac Soc. 8:223–72. 2011. View Article : Google Scholar : PubMed/NCBI
26	Ortiz LA, Gambelli F, McBride C, Gaupp D, Baddoo M, Kaminski N and Phinney DG: 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
27	Hansmann G, Fernandez-Gonzalez A, Aslam M, Vitali SH, Martin T, Mitsialis SA and Kourembanas S: Mesenchymal stem cell-mediated reversal of bronchopulmonary dysplasia and associated pulmonary hypertension. Pulm Circ. 2:170–81. 2012. View Article : Google Scholar : PubMed/NCBI
28	Jun D, Garat C, West J, Thorn N, Chow K, Cleaver T, Sullivan T, Torchia EC, Childs C, Shade T, et al: The pathology of bleomycin-induced fibrosis is associated with loss of resident lung mesenchymal stem cells that regulate effector T cell proliferation. Stem Cells. 29:725–735. 2011. View Article : Google Scholar : PubMed/NCBI
29	Pan MG, Xiong Y and Chen F: NFAT gene family in inflammation and cancer. Curr Mol Med. 13:543–554. 2013. View Article : Google Scholar : PubMed/NCBI
30	Kudryavtseva O, Aalkjaer C and Matchkov VV: Vascular smooth muscle cell phenotype is defined by Ca²⁺-dependent transcription factors. FEBS J. 280:5488–5499. 2013. View Article : Google Scholar : PubMed/NCBI
31	Koulmann N, Novel-Chaté V, Peinnequin A, Chapot R, Serrurier B, Simler N, Richard H, Ventura-Clapier R and Bigard X: Cyclosporin A inhibits hypoxia-induced pulmonary hypertension and right ventricle hypertrophy. Am J Respir Crit Care Med. 174:699–705. 2006. View Article : Google Scholar : PubMed/NCBI