The MODULATORY EFFECTS OF BONE MARROW-DERIVED MESENCHMAL STEM CELLS ON CYCLOPHOSPHAMIDE INDUCED HEPATOTOXICITY IN CARCINOMA MICE
Keywords:Hepatotoxicity, Ehrlich, Oxidative stress, Inflammation, Apoptosis
Objective: Cyclophosphamide (CPA) is a chemotherapeutic agent, induces hepatotoxicity as one of its side effects. Therefore, the present work was designed to investigate the protective role of bone marrow-derived mesenchymal stem cells (BM-MSCs) on CPA–induced hepatotoxicity in Ehrlich Ascites Carcinoma bearing-mice (EAC) and to test whether BM-MSCs influences the antitumor properties of the CPA.
Methods: The hepatoprotective effects of BM-MSCs (single dose of 100 µl of a cell suspension containing allogenic BM-MSCs, i. v.) was evaluated in a model of hepatotoxicity by CPA (10 mg/kg/d i. p.) in EAC-female mice for one month. The anti-tumor activities of CPA and BM-MSCs were assessed by measuring mean tumor weight, mean survival time and the increase in life span. Moreover, ALT, AST, GGT, MDA, GSH, SOD, IL-6, IL10, caspase-3 and Bcl2 were measured.
Results: The i. p. administration of CPA and BM-MSCs resulted in significant reductions in tumor size and mean tumor weight as well as caused concurrent significant increases in the life span as compared to the EAC mice. Furthermore, BM-MSCs ameliorated the liver enzyme markers namely ALT, AST, GGT, and hepatic oxidative stress through inhibition of MDA level that correlated with significant improvement in antioxidant status via increasing GSH and SOD levels as compared to both EAC and EAC+CPA groups. Moreover, BM-MSCs treatment significantly reduced the inflammatory marker level IL-6 as well as increment the level of IL-10 with subsequent decreases apoptosis via a depletion in the caspase-3 associated with an enhancement in the level of Bcl2 as compared to EAC group and EAC+CPA group. Minor histological lesions were observed in the liver tissue sections of mice treated with CPA and BM-MSCs as compared to the high histological lesions observed in the liver of the EAC group and CPA treated group.
Conclusion: These results concluded that the combination treatment of BM-MSCs with CPA exhibited promising potential antitumor efficacy with greater safety than CPA treatment alone in mice via its antioxidant, anti-inflammatory and antiapoptotic effects.
Abbas S, Malla S. Cytotoxicity and expression studies of angiogenesis-promoting genes in cancer cell lines under the treatment of cancer candidate drugs. Asian J Pharm Clin Res 2019;12:130-4.
Barakat W, Elshazly SM, Mahmoud AA. Spirulina platensis lacks antitumor effect against Solid Ehrlich Carcinoma in female mice. Adv Pharmacol 2015. Doi:10.1155/2015/132873
Sinanoglu O, Yener AN, Ekici S, Midi A, Aksungar FB. The protective effects of spirulina in cyclophosphamide induced nephrotoxicity and urotoxicity in rats. Urology 2012;80:1392-6.
Jiang W, Liu J, Li P, Lu Q, Pei X, Sun Y, et al. Magnesium iso glycyrrhizinate shows hepatoprotective effects in a cyclophosphamide induced model of hepatic injury. Oncotarget 2017;8:33252-64.
Kim J, Chen CH, Yang J, Mochly Rosen D. Aldehyde dehydrogenase 2*2 knock-in mice show increased reactive oxygen species production in response to cisplatin treatment. J Biomed Sci 2017;24:33.
Mahmoud AM, Germoush MO, Alotaibi MF, Hussein OE. Possible involvement of Nrf2 and PPARγ up-regulation in the protective effect of umbelliferone against cyclophosphamide-induced hepatotoxicity. Biomed Pharmacother 2017;86:297–306.
Subramaniam SR, Cader RA, Mohd R, Yen KW, Ghafor HA. Low-dose cyclophosphamide-induced acute hepatotoxicity. Am J Case Rep 2013;14:345.
Shokrzadeh M, Ahmadi A, Naghshvar F, Chabra A, Jafarinejhad M. Prophylactic efficacy of melatonin on cyclophosphamide-induced liver toxicity in mice. Bio Med Res Int 2014. https://doi.org/10.1155/2014/470425
Jalali AS, Hasanzadeh S, Malekinejad H. Achillea millefolium inflorescence aqueous extract ameliorates cyclophosphamide-induced toxicity in rat testis: stereological evidences. Chin J Nat Med 2012;1:247–54.
Abd El-Fattah AI, Zaghloul MS, Eltablawy NA, Rashed LA. a-Lipoic acid and amlodipine/perindopril combination potentiate the therapeutic effect of mesenchymal stem cells on isoproterenol induced cardiac injury in rats. Biochimie 2019;156:59-68.
Caplan AL, Dennis JE. Mesenchymal stem cells as trophic mediators. J Cell Biochem 2006;98:1076–84.
Vincent PC, Nicholls A. Comparison of the growth of the Ehrlich ascites tumor in male and female mice. Cancer Res 1967;27:1058–65.
Strober W. Trypan blue exclusion test of cell viability. Curr Protoc Immunol 2001;21:A.3B.1-2.
Abdel Aziz M, Atta H, Mahfouz S, Fouad H, Roshdy N, Ahmed H, et al. Therapeutic potential of bone marrow derived mesenchymal stem cells on experimental liver fibrosis. Clin Biochem 2007;40:893-9.
Jaiswal N, Haynesworth S, Caplan A, Bruder S. Osteogenic differentiation of purified, culture-expanded human mesenchymal stem cells in vitro. J Cell Biochem 1997;64:295-312.
Perry MJ. The chemotherapy source book. Wolters Kluwer Health/lippincott Williams and Wilkins. Philadelphia; 2008.
Agrawal SS, Saraswati S, Mathur R, Pandey M. Cytotoxic and antitumor effects of brucine on Ehrlich ascites tumour and human cancer cell line. Life Sci 2011;89:147-58.
El Gammal ZH, Rashed LA, Abdel Aziz MT, Ahmed H, Elwahy M, Youakim MF, et al. Comparative study between the attenuation of cardiac fibrosis by mesenchymal stem cells versus colchicine. Acta Med Int 2016;3:137.
Fahim FA, Esmat AY, Mady EA, Ibrahim EK. Antitumor activities of Iodo acetate and Di methyl sulphoxide against solid Ehrlich carcinoma growth in mice. Biol Res 2003;36:253-62.
Lowry OH, Rosebrough NJ, Farr AL, Randall RJ. Protein measurement with the Folin phenol reagent. J Biol Chem 1951;193:265-75.
Young DS. Effect of drugs on clinical laboratory tests. 3rd ed. Washington DC: AACC Press; 1990. p. 6-12.
Satoh K. Serum lipid peroxide in cerebrovascular disorders determined by a new colorimetric method. Clin Chim Acta 1978;90:37-43.
Beutlr E, Duron O, Kelly BM. Improved method for the determination of blood glutathione. J Lab Clin Med 1963;61:882-8.
Nishikimi M, Appaji N, Yagi K. The occurrence of superoxide anion in the reaction of reduced phenazine methosulfate and molecular oxygen. Biochem Biophys Res Commun 1972;46:849-54.
Barbareschi M, Veronese S, Leek R, Fox S, Bonzanini M, Girlando S. Bcl-2 and P53 expression in node-negative breast carcinoma-a study with long-term follow up. Hum Pathol 1996;27:1149–55.
Banchroft JD, Stevens A, Turner DR. Theory and practice of histological techniques. 4th ed. New York, London, San Francisco, Tokyo: Churchil Livingstone; 1996.
Abramoff MD, Magalhaes PJ, Ram SJ. Image processing with image. J Biophotonics Int 2004;11:36-42.
Agrawal SS, Sharma P. Anticancer activity of cyclophosphamide nanoparticles against ehrlich ascites carcinoma cells bearing swiss albino mice. Inter J Pharm Pharm Res 2017;9:244-65.
Mahipal P, Pawar RS. Nephroprotective effect of Murraya koenigii on cyclophosphamide induced nephrotoxicity in rats. Asian Pac J Trop Med 2017;10:869–73.
Han X, Yang Q, Lin L, Xu C, Zheng C, Chen X, et al. Interleukin-17 enhances immunosuppression by mesenchymal stem cells. Cell Death Differentiation 2014;21:1758–68.
Su J, Chen X, Huang Y, Li W, Li J, Cao K, et al. Phylogenetic distinction of iNOS and IDO function in mesenchymal stem cell-mediated immunosuppression in mammalian species. Cell Death Differentiation 2014;21:388–96.
Chakraborty AP, Singha Roya PA, Basua A, Bhattacharyaa S. Sensitization of cancer cells to cyclophosphamide therapy by an organo-selenium compound through ROS-mediated apoptosis. Biomed Pharmacother 2016;841:992–9.
Zheng Y, Yang F, Fu L, Liu K. The mechanism of miR-143 inducing apoptosis of liver carcinoma cells through regulation of the NF-kappaB pathway. Oncol Lett 2018;15:9567-71.
Donia TIK, Gerges MN, Mohamed TM. Amelioration effect of Egyptian sweet orange hesperidin on Ehrlich ascites carcinoma (EAC) bearing mice. Chem Biol Interact 2018;285:76–84.
Apostolou G, Papanikolaou IG, Katselis C, Feretis T, Kletsas D, Konstadoulakis MM, et al. Undifferentiated adipose tissue stem cell transplantation promotes hepatic regeneration, ameliorates histopathologic damage of the liver, and upregulates the expression of liver regeneration-and liver-specific genes in a rat model of partial hepatectomy. Stem Cells Int 2018;1–18. Doi:10.1155/2018/1393607
Maruthanila VL, Poornima J, Mirunalini S. Attenuation of carcinogenesis and the mechanism underlying by the influence of indole-3-carbinol and its metabolite 3, 3′-diindolylmethane: a therapeutic marvel. Adv Pharmacol Sci 2014. Doi:10.1155/2014/832161
Iyer SS, Torres Gonzalej E, Neujahr DC, Kwon M, Brigham KL, Jones DP, et al. Effect of bone marrow-derived mesenchymal stem cells on endotoxin-induced oxidation of plasma cysteine and glutathione in mice. Stem Cell Int 2010. https://doi.org/10.4061/2010/868076
Zhang M, Li DS, Chang L, Zhang Y, Liu R, Sun F, et al. Bone marrow mesenchymal stem cell transplantation retards the natural senescence of rat hearts. Stem Cell Transl Med 2015;4:494.
Willing AE, Lixian J, Milliken M, Poulos S, Zigova T, Song S, et al. Intravenous versus intrastriatal cord blood administration in a rodent model of stroke. J Neurosci Res 2003;73:296.
Waldner MJ, Foersch S, Neurath MF. Interleukin-6-a key regulator of colorectal cancer development. Int J Biol Sci 2012;8:1248-53.
Grivennikov SI, Karin M. Inflammatory cytokines in cancer: tumour necrosis factor and interleukin 6 take the stage. Ann Rheum Dis 2011;70:i104-8.
Blankenstein T. The role oftumor stroma in the interaction between tumor and immune. Curr Opin Immunol 2005;17:180–6.
Mocellin S, Marincola FM, Young HA. Interleukin-10 and the immune response against cancer: a counterpoint. J Leukoc Biol 2005;78:1043–51.
Moore KW, de Waal Malefyt R, Coffman RL, O’Garra A. Interleukin-10 and the interleukin-10 receptor. Annu Rev Immunol 2001;19:683–765.
Gazzinelli RT, Oswald IP, James SL, Sher A. IL-10 inhibits parasite killing and nitrogen oxide production by IFN-gamma-activated macrophages. J Immunol 1992;148: 1792–6.
Lechner M, Lirk P, Rieder J. Inducible nitric oxide synthase (iNOS) in tumor biology: the two sides of the same coin. Semin Cancer Biol 2005;15:277–89.
Trombetta D, Cimino F, Cristani M, Mandalari G, Saija A, Ginestra G, et al. In vitro protective effects of two extracts from bergamot peels on human endothelial cells exposed to tumor necrosis factor-a (TNF-a). J Agric Food Chem 2010;58:8430-6.
Kumar S, Dhankhar N, Kar V, Shrivastava M, Shrivastava S. Myocardial injury provoked by cyclophosphamide, protective aspect of hesperidin in rats. Int J Res Pharm Biomed Sci 2011;2:1288-96.
Bryniarski K. The influence of cyclophosphamide on immune function of murine macrophages. Pharmacology 2012;143-60. Doi:10.5772/32891
Carrero R, Cerrada I, Lledo E, Dopazo J, Garcia Garcia F, Rubio M, et al. IL1β induces mesenchymal stem cells migration and leucocyte chemotaxis through NF-κβ. Stem Cell Rev Rep 2012;8:905–16.
Liang X, Ding Y, Zhang Y, Tse HF, Lian Q. Paracrine mechanisms of mesenchymal stem cell-based therapy: current status and perspectives. Cell Transplant 2014;23:1045.
You P, Xing F, Huo J, Wang B, Di J, Zeng S, et al. In vitro and in vivo evaluation of anisomycin against Ehrlich ascites carcinoma. Oncol Rep 2013;29:2227–36.
Zhang S, Yang Y, Liang Z, Duan W, Yang J, Yan J, et al. Silybin-mediated inhibition of Notch signaling exerts antitumor activity in human hepatocellular carcinoma cells. PLOS ONE 2013;8:e83699.
Da Mota MF, Benfica PL, Batista AC, Martins FS, de Paula JR, Valadares MC. Investigation of Ehrlich ascites tumor cell death mechanisms induced by Synadenium umbellatum. Pax J Ethnopharmacol 2012;139:319–29.
Badr El-Din NK, Ali DA, Abou-El-Magd RF. Grape seeds and skin induce tumor growth inhibition via G1-phase arrest and apoptosis in mice inoculated with Ehrlich ascites carcinoma. Nutrition 2019;58:100-9.
Brentnall M, Rodriguez Menocal L, De Guevara RL, Cepero E, Boise LH. Caspase-9, caspase-3 and caspase-7 have distinct roles during intrinsic apoptosis. BMC Cell Biol 2013;14:32.
Xu LX, Yan LJ, Huang SP. Ganoderic acid A against cyclophosphamide induced hepatic toxicity in mice. J Biochem Mol Toxicol 2019;33:e22271.
Chimenti I, Smith RR, Li TS, Gerstenblith G, Messina E, Giacomello A, et al. An relative roles of direct regeneration versus paracrine effects of human cardiosphere-derived cells transplanted into infarcted mice. Circ Res 2010;106:971.
Hirsch E, Hilfiker Kleiner D, Balligand JL. Interaction of the heart and its close and distant neighbours: report of the meeting of the esc working groups myocardial function and cellular biology. Cardiovasc Res 2013;99:595.
Feng Y, Huang W, Wani M. Ischemic preconditioning potentiates the protective effect of stem cells through secretion of exosomes by targeting Mecp2 via miR-22. PloS One 2014;9:e88685.
Ali DA, Badr El-Din NK, Abou-El-magd RF. Antioxidant and hepatoprotective activities of grape seeds and skin against Ehrlich solid tumor induced oxidative stress in mice. Egyptian J Basic Appl Sci 2015;2:98-109.
Rahmouni F, Hamdaoui L, Badraoui R, Rebai T. Protective effects of Teucrium polium aqueous extract and ascorbic acid on hematological and some biochemical parameters against carbon tetrachloride (CCl4) induced toxicity in rats. Biomed Pharmacother 2017;9:43–8.
Chen Q, Zhouc R, Zhanga Y, Zhua S, Xiaob C, Gonge J, et al. Bone marrow mesenchymal stromal cells attenuate liver allograft rejection may via upregulation PD-L1 expression through downregulation of miR-17-5p. Transplant Immunol 2018;51:21–9.