Effects of azathioprine and infliximab on mesenchymal stem cells derived from the bone marrow of rats in vitro

Huang,Hua‑Rong; Zan,Hui; Lin,Ying; Zhong,Ying‑Qiang

doi:10.3892/mmr.2014.1905

Effects of azathioprine and infliximab on mesenchymal stem cells derived from the bone marrow of rats in vitro

Authors:
- Hua‑Rong Huang
- Hui Zan
- Ying Lin
- Ying‑Qiang Zhong
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Affiliations: Department of Pediatrics, Sun Yat‑sen Memorial Hospital, Sun Yat‑sen University, Guangzhou, Guangdong 510120, P.R. China, Department of Gastroenterology and Hepatology, Sun Yat‑sen Memorial Hospital, Sun Yat‑sen University, Guangzhou, Guangdong 510120, P.R. China
Published online on: January 17, 2014 https://doi.org/10.3892/mmr.2014.1905
Pages: 1005-1012

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Abstract

Mesenchymal stem cell (MSC) transplantation has been demonstrated to be promising in the treatment of inflammatory bowel disease (IBD). Azathioprine (AZA) is widely used in IBD patients. Infliximab, as a representative of biological therapy for IBD, is important in the treatment regimen. In the present study we investigated the effects of AZA and infliximab on the cell proliferation, cell cycle and apoptosis of the MSCs derived from the bone marrow of Sprague‑Dawley (SD) rats in vitro in order to provide preliminary data for optimizing the treatment of IBD. MSCs derived from the bone marrow of rats were either cultured in various concentrations of AZA‑ or infliximab‑supplemented medium for 24, 48 and 72 h, respectively. The growth curves of MSCs were obtained. The apoptosis and the cell cycle of the MSCs were analyzed by flow cytometry. AZA decreased the proliferation of MSCs by 66% and increased apoptosis at 0.20 mg/ml for 72 h (P<0.05). The percentage of necrotic cells increased markedly in MSCs treated with 0.30 mg/ml AZA for 72 h (P<0.05). As the exposure time increased, the percentage of MSCs in phase G0‑G1 increased and that in phase S decreased in AZA groups exceeding 0.20 mg/ml (P<0.05). However, infliximab had a minimal impact on the cell proliferation, apoptosis and cell cycle of the MSCs. AZA was able to inhibit cell proliferation and induce apoptosis of the MSCs in vitro. Infliximab did not affect the cell proliferation, apoptosis and cell cycle of the MSCs derived from rats.

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1	Beyer Nardi N and da Silva Meirelles L: Mesenchymal stem cells: isolation, in vitro expansion and characterization. Handb Exp Pharmacol. 174:249–282. 2006.
2	Li YP, Paczesny S, Lauret E, et al: Human mesenchymal stem cells license adult CD34⁺ hemopoietic progenitor cells to differentiate into regulatory dendritic cells through activation of the Notch pathway. J Immunol. 180:1598–1608. 2008.PubMed/NCBI
3	Powell DW, Mifflin RC, Valentich JD, Crowe SE, Saada JI and West AB: Myofibroblasts. II Intestinal subepithelial myofibroblasts. Am J Physiol. 277:C183–C201. 1999.PubMed/NCBI
4	Lazebnik LB, Konopliannikov AG, Kniazew OV, et al: Use of allogeneic mesenchymal stem cells in the treatment of intestinal inflammatory disease. Ter Arkh. 82:38–43. 2010.(In Russian).
5	Gonzalez-Rey E, Anderson P, Gonzalez MA, Rico L, Büscher D and Delgado M: Human adult stem cells derived from adipose tissue protect against experimental colitis and sepsis. Gut. 58:929–939. 2009. View Article : Google Scholar : PubMed/NCBI
6	Suarez-Almazor ME, Spooner C and Belseck E: Azathioprine for treating rheumatoid arthritis. Cochrane Database Syst Rev. 4:CD0014612000.PubMed/NCBI
7	Sandborn WJ: Azathioprine: state of the art in inflammatory bowel disease. Scand J Gastroenterol. (Suppl 225): 92–99. 1998. View Article : Google Scholar
8	Patel AA, Swerlick RA and McCall CO: Azathioprine in dermatology: the past, the present, and the future. J Am Acad Dermatol. 55:369–389. 2006. View Article : Google Scholar : PubMed/NCBI
9	Rumbo C, Emerick KM, Emre S and Shneider BL: Azathioprine metabolite measurements in the treatment of autoimmune hepatitis in pediatric patients: a preliminary report. J Pediatr Gastroenterol Nutr. 35:391–398. 2002. View Article : Google Scholar : PubMed/NCBI
10	Abu-Shakra M and Shoenfeld Y: Azathioprine therapy for patients with systemic lupus erythematosus. Lupus. 10:152–153. 2001. View Article : Google Scholar : PubMed/NCBI
11	Tiede I, Fritz G, Strand S, et al: CD28-dependent Rac1 activation is the molecular target of azathioprine in primary human CD4⁺ T lymphocytes. J Clin Invest. 111:1133–1145. 2003. View Article : Google Scholar : PubMed/NCBI
12	Waters OR and Lawrance IC: Understanding the use of immunosuppressive agents in the clinical management of IBD. Curr Drug Targets. 12:1364–1371. 2011. View Article : Google Scholar : PubMed/NCBI
13	Chevaux JB, Peyrin-Biroulet L and Sparrow MP: Optimizing thiopurine therapy in inflammatory bowel disease. Inflamm Bowel Dis. 17:1428–1435. 2011. View Article : Google Scholar : PubMed/NCBI
14	Oh CJ, Das KM and Gottlieb AB: Treatment with anti-tumor necrosis factor alpha (TNF-alpha) monoclonal antibody dramatically decreases the clinical activity of psoriasis lesions. J Am Acad Dermatol. 42:829–830. 2000. View Article : Google Scholar : PubMed/NCBI
15	Maini R, St Clair EW, Breedveld F, et al: Infliximab (chimeric anti-tumour necrosis factor alpha monoclonal antibody) versus placebo in rheumatoid arthritis patients receiving concomitant methotrexate: a randomised phase III trial. ATTRACT Study Group. Lancet. 354:1932–1939. 1999. View Article : Google Scholar
16	Braun J and Sieper J: Ankylosing spondylitis. Lancet. 369:1379–1390. 2007. View Article : Google Scholar
17	Yamamoto-Furusho JK: Current and future biological treatments in inflammatory bowel disease. Gene Therapy Applications. Kang C: InTech; Rijeka: pp. 363–374. 2011
18	Van den Brande JM, Braat H, van den Brink GR, et al: Infliximab but not etanercept induces apoptosis in lamina propria T-lymphocytes from patients with Crohn’s disease. Gastroenterology. 124:1774–1785. 2003.
19	Efron PA and Moldawer LL: Cytokines and wound healing: the role of cytokine and anticytokine therapy in the repair response. J Burn Care Rehabil. 25:149–160. 2004. View Article : Google Scholar : PubMed/NCBI
20	Böcker W, Docheva D, Prall WC, et al: IKK-2 is required for TNF-alpha-induced invasion and proliferation of human mesenchymal stem cells. J Mol Med (Berl). 86:1183–1192. 2008.PubMed/NCBI
21	Duijvestein M, Molendijk I, Roelofs H, et al: Mesenchymal stromal cell function is not affected by drugs used in the treatment of inflammatory bowel disease. Cytotherapy. 13:1066–1073. 2011. View Article : Google Scholar : PubMed/NCBI
22	Zan H and Zhong YQ: The progress of mesenchymal stem cells in the treatment of inflammatory bowel disease. New Medicine. 4:211–215. 2011.(In Chinese).
23	Fischer UM, Harting MT, Jimenez F, et al: Pulmonary passage is a major obstacle for intravenous stem cell delivery: the pulmonary first-pass effect. Stem Cells Dev. 18:683–692. 2009. View Article : Google Scholar : PubMed/NCBI
24	Schrepfer S, Deuse T, Reichenspurner H, Fischbein MP, Robbins RC and Pelletier MP: Stem cell transplantation: the lung barrier. Transplant Proc. 39:573–576. 2007. View Article : Google Scholar
25	Lee RH, Pulin AA, Seo MJ, et al: Intravenous hMSCs improve myocardial infarction in mice because cells embolized in lung are activated to secrete the anti-inflammatory protein TSG-6. Cell Stem Cell. 5:54–63. 2009. View Article : Google Scholar : PubMed/NCBI
26	Lazebnik LB, Kniazev OV, Konopliannikov AG, et al: Allogeneic mesenchymal stromal cells in patients with ulcerative colitis: two years of observation. Eksp Klin Gastroenterol. 11:3–15. 2010.(In Russian).
27	Baert F, Noman M, Vermeire S, Van Assche G, D’Haens G, Carbonez A and Rutgeerts P: Influence of immunogenicity on the long-term efficacy of infliximab in Crohn’s disease. N Engl J Med. 348:601–608. 2003.
28	Sagynbaeva VÉ, Lazebnik LB, Kniazev OV and Efremov LI: Antibodies to infliximab and antigens HLA I–II class as the witnesses of immune response to the biological treatment of inflammatory bowel disease. Eksp Klin Gastroenterol. 12:7–14. 2011.(In Russian).