Effects of Cyclosporine and Azacitidine on Some Hematologic and ‎Biochemical Parameters of Benzene-Induced Aplastic Anemia in Rats

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Ghanem Taher
Huda F Hasn


Aplastic anemia, marked by deficiencies in hematopoietic stem cells, leads to peripheral blood pancytopenia and hypocellular bone ‎marrow. This study aimed to evaluate the therapeutic efficacy of cyclosporine and azacitidine, administered either alone or in combination, in rats with benzene-induced aplastic ‎anemia, focusing on restoring normal blood cell levels and preventing disease complications. Thirty adult female Wistar rats ‎(Rattus ‎norvegicus)‎ were randomly divided into five groups: negative control (C-, untreated), positive control (C+, induced aplastic anemia with ‎distilled water), cyclosporine-treated (CsA, 5.86 mg/kg), azacitidine-treated (Aza, 5.75 mg/kg), and combination-treated (CsA+Aza, 3.68 ‎mg/kg each). Benzene (1940 mg/kg) was administered orally for fifteen days to induce aplastic anemia. Post a 30-day treatment period, ‎evaluations included differential WBC and reticulocyte counts, serum IL-2 levels, and alkaline phosphatase (ALP) activity. Results ‎showed significant improvements in WBC% and reticulocyte% in all treated groups compared to the C+ group, with the combination-‎treated group showing the highest enhancement. IL-2 levels in the combination group were significantly reduced compared to other ‎treatment groups, aligning closely with the negative control. The ALP activity was significantly higher in both the cyclosporine and ‎azacitidine-treated groups compared to the positive control, with the combination group showing a marked increase over the azacitidine ‎group but no significant difference from the cyclosporine group and negative control.‎ In conclusion, the study demonstrates the potential therapeutic benefits of cyclosporine and azacitidine in treating benzene-induced ‎aplastic anemia in rats. The combination therapy, in particular, showed improved efficacy in all tested parameters, suggesting a potential ‎strategy for dose reduction and toxicity mitigation.


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Effects of Cyclosporine and Azacitidine on Some Hematologic and ‎Biochemical Parameters of Benzene-Induced Aplastic Anemia in Rats. (2023). The Iraqi Journal of Veterinary Medicine, 47(2), 9-14. https://doi.org/10.30539/ijvm.v47i2.1455

How to Cite

Effects of Cyclosporine and Azacitidine on Some Hematologic and ‎Biochemical Parameters of Benzene-Induced Aplastic Anemia in Rats. (2023). The Iraqi Journal of Veterinary Medicine, 47(2), 9-14. https://doi.org/10.30539/ijvm.v47i2.1455


Sweeney R, Esmail F, Mirza KM, Nand S. Hypercellular bone marrow in aplastic anemia: A case report of two patients. Clin Case Rep. 2021;9(11):e04845. https://doi.org/10.1002/ccr3.4845

AL-Jaff SHK. Anemia and low testosterone associated with male type 2 diabetic patients. Iraqi ‎J. Vet. Med. 2010;34(2):58-65.‎‏‎ ‎ https://doi.org/10.30539/iraqijvm.v34i2.632

Peffault de Latour R, Kulasekararaj A, Iacobelli S, Terwel SR, Cook R, Griffin M, et al. ‎Eltrombopag added to immunosuppression in severe aplastic anemia. N Engl J Med. ‎‎2022;386(1):11-23.


Segel GB, Lichtman ML. Aplastic Anemia: Acquired and Inherited. Williams Hematology, 9Ed. ‎McGraw-Hill Education/Medical. Chapter. ‎‎2015; 34.‎

Abdulrazzaq GT, Hasan HF. Role of cyclosporine and azacitidine on the treatment of induced ‎aplastic anemia in female rats. Eur. Chem. ‎Bull. 2023; 12(1): 2579-2594.


‎Javan M R, Saki N, Moghimian‐Boroujeni B. Aplastic anemia, cellular and molecular aspects. ‎Cell Biol. Int. 2021; 45(12): 2395-2402.‎‏‎ ‎ https://doi.org/10.1002/cbin.11689

‎‎Bouzid D, Fourati H, Amouri A, Marques I, Abida O, Tahri N, et al. Autoimmune diseases ‎association study with the KIAA1109-IL2-IL21 ‎region in a Tunisian population. Mol. Biol. Rep. ‎‎2014; 41(11): 7133-7139. https://doi.org/10.1007/s11033-014-3596-5

De R Dutta A, Dolai TK, Ghosh K, Halder A. Comparative study of bone marrow and blood ‎plasma levels of IL-2 in aplastic anaemia and ‎their relationship with disease severity. ‎Hematology. 2019; 24(1): 84-88.‎‏‎ https://doi.org/10.1080/10245332.2018.1512391

‎Ershad A, Taziki S, Ebrahimian, M, Abadi SSD. Acute cyclosporine overdose: A systematic ‎review. Med. Clin. Pract. 2023; 6(2): 100358.‎‏‎ https://doi.org/10.1016/j.mcpsp.2022.100358

Bianco CM, Robinson MR. Chronic immunosuppression medications. Contemporary Heart ‎Transplantation. 2020; 251-283.‎‏‎ https://doi.org/10.1007/978-3-319-58054-8_23

Mehdipour P, Murphy T, De Carvalho DD. The role of DNA-demethylating agents in cancer ‎therapy. Pharmacol. Ther. 2020;205:107416.‎‏‎ https://doi.org/10.1016/j.pharmthera.2019.107416

Abdulrazzaq GT, Hasan HF. Evaluation the pharmacodynamics interaction of cyclosporine ‎and azacitidine on the treatment of induced ‎aplastic anemia in rats. MINAR Int. J. Appl. Sci. ‎Tech. 2023. In press. https://doi.org/10.47832/MinarCongress9-24

Ata SIE. Induction of aplastic anemia in experimental model. Int J Appl Biol Pharm Tech. ‎‎2016; 7(1):182-195.

Hasan HF, Krair EM, Al-Halbosy MF. Evaluating Anti-Anemic Effect of Mesenchymal Stem ‎Cells and Oxymetholone on Aplastic Anemia ‎Induced in Mice.‎‏ ‏‎ Indian J. Sci. 2017;8(44):12714-‎‎12729.‎

Lewis JJ, West HF, Sundaram V, Caldwell SH. The Prevalence of spur cell anemia in ‎hospitalized cirrhotics and its relationship to ‎coagulation, hemolysis and HDL Cholesterol:‎‎322. Am. J. Gastroenterol. 2006;101:S151. https://doi.org/10.14309/00000434-200609001-00322

Alkhedaide AQ. Anti-inflammatory Effect of Juniperus Procera Extract in Rats Exposed to ‎Streptozotocin Toxicity. Anti-Inflamm. Anti-‎Allergy Agents Med. Chem. 2019;18:71-79.


Alkhedaide AQ. Anti-inflammatory effect of Juniperus procera extract in rats exposed to streptozotocin toxicity. Antiinflamm Antiallergy Agents Med Chem. 2019;18(1):71-79.


Lens S, Calleja JL, Campillo A, Carrión JA, Broquetas T, Perello C, Forns X. Aplastic anemia ‎and severe pancytopenia during treatment with ‎peg-interferon, ribavirin and telaprevir for chronic ‎hepatitis C. WJG, World J Gastroenterol. 2015;21(17):5421.‎‏‎ https://doi.org/10.3748/wjg.v21.i17.5421

Adeyemi OT, Osilesi O, Adebawo OO, Onajobi FD, Oyedemi SO, Afolayan AJ. Alkaline ‎phosphatase (ALP), aspartate aminotransferase (AST) ‎and alanine aminotransferase (ALT) ‎Activities in selected tissues of rats fed on processed atlantic horse mackerel (Trachurus ‎trachurus). ‎J. Adv. Biol. 2015;6(03):139.

‎‏‎‏ https://doi.org/10.4236/abb.2015.63014

Taher AG, Hasan, HF. Antagonistic effect of Avena sativa seeds crude extract to ‎chlorambucil effect on fertility of female rats. Biochem. ‎Cell. Arch. 2019; 19(2):3363-3370.‎

Al-Naimi RA. Hematological and Biochemical evaluation after different orally doses of ‎Copper sulfate in rats. Iraqi J. Vet. Med. 2014; ‎‎38(1):83-91. https://doi.org/10.30539/iraqijvm.v38i1.259

De Bruin AM, Voermans C, Nolte MA. Impact of interferon-γ on hematopoiesis. Blood, Am. J. ‎Hematol. 2014;124(16):2479-2486. https://doi.org/10.1182/blood-2014-04-568451

Kakiuchi T, Eguchi K, Koga D, Eguchi H, Nishi M, Sonoda M, Ishimura M, Matsuo M. Changes in bone marrow and peripheral blood lymphocyte subset findings with onset of hepatitis-associated aplastic anemia. Medicine (Baltimore). 2022;101(8):e28953. https://doi.org/10.1097/MD.0000000000028953

Perry SS, Kim M, Spangrude GJ. Direct effects of cyclosporin A on proliferation of hematopoietic stem and progenitor cells. Cell Transplant. 1999;8(4):339-344. https://doi.org/10.1177/096368979900800401

Elsayed ASI, Jbireal J, Azab AE. Effect of acute and chronic cyclosporine A treatment on ‎haematological data in male albino rats. J Appl ‎Biotechnol Bioeng. 2018;5(6):350-357.


Boada M, Echarte L, Guillermo C, Diaz L, Touriño C, Grille S. 5-Azacytidine restores ‎interleukin 6-increased production in mesenchymal ‎stromal cells from myelodysplastic patients. ‎Hematol Transfus Cell Ther. 2021;43:35-42. https://doi.org/10.1016/j.htct.2019.12.002

Gilmartin A G. Groy A, Gore ER, Atkins C, Long ER, Montoute MN, Benowitz AB. In vitro and ‎in vivo induction of fetal hemoglobin with a ‎reversible and selective DNMT1 inhibitor. ‎Haematologica. 2021;106(7):1979. https://doi.org/10.3324/haematol.2020.248658‎

Boyman O, Sprent J. The role of interleukin-2 during homeostasis and activation of the ‎immune system. Nat. Rev. Immunol, 2012;12(3): ‎‎180-190. https://doi.org/10.1038/nri3156

Damoiseaux J. The IL-2-IL-2 receptor pathway in health and disease: The role of the soluble ‎IL-2 receptor. Clin. Immunol. 2020;218: ‎‎108515. https://doi.org/10.1016/j.clim.2020.108515

Russell G, Graveley R, Seid J, Al-Humidan, AK, Skjodt H. Mechanisms of action of ‎cyclosporine and effects on connective tissues. In ‎Seminars in arthritis and rheumatism. 1992;‎‎21(6):16-22. WB Saunders.‎‏‎ https://doi.org/10.1016/0049-0172(92)90009-3

Satake A, Schmidt AM, Nomura S, Kambayashi T. Inhibition of calcineurin abrogates while inhibition of mTOR promotes regulatory T cell expansion and graft-versus-host disease protection by IL-2 in allogeneic bone marrow transplantation. PLoS One. 2014;9(3):e92888. https://doi.org/10.1371/journal.pone.0092888

Chen H, Li J, Wang Q. Associations between bone-alkaline phosphatase and bone mineral ‎density in adults with and without diabetes. ‎Medicine. 2018;97(17):e0432.‎‏‎‏


Sanchez-Abarca LI, Gutierrez-Cosio S, Santamaría C, Caballero-Velazquez T, Blanco B, ‎Herrero-Sánchez C, et al. Immunomodulatory effect ‎of 5-azacytidine (5-azaC): potential role in ‎the transplantation setting. Blood, Am. J. Hematol 2010;115(1):107-121. https://doi.org/10.1182/blood-2009-03-210393

Dowling MR, Kan A, Heinzel S, Marchingo JM, Hodgkin PD, Hawkins ED. Regulatory T cells ‎suppress effector T cell proliferation by ‎limiting division destiny. Front Immunol. 2018;9:2461.


Calvi M, Adams GB, Weibrecht KW, Weber JM, Olson DP, Knight MC,et al . Osteoblastic ‎cells regulate the haematopoietic stem cell niche. ‎Nature. 2003;425(6960):841-846. https://doi.org/10.1038/nature02040

Ali MR, Zaidan TF, Gorial, FI. Validity of osteocalcin and alkaline phosphatase biomarkers in ‎postmenopausal women with low bone ‎mineral density. Chem Mat Res. 2014;6(3): 13-19.‎

Hassan SL. Toxic pathological changes on albino mice after exposures to cypermethrin. ‎‎2019;9(52):16348-16354.‎

Al-Ameedi AI, Ali l, AL-Rekabi FM, Al-Rikabi ShJ. Hepatotoxic effect of chronic ‎exposure of Tacrolimus in male Albino rats. ‎Iraqi J. Vet. Med. 2016;40(1):161-166. https://doi.org/10.30539/iraqijvm.v40i1.155

Essa EA, AL-Sharqi SA, Yenzeel JH, Mohammed LJ. Histological and enzyme histochemical ‎changes of liver induced by the action of ‎maxxthor insecticide in male albino rats. Med. Legal ‎Update, 2020;20(3):1047-1053.‎

Atmar K, Tulling AJ, Lankester AC, Bartels M, Smiers FJ, van der Burg M, et al. Functional and Immune Modulatory Characteristics of Bone Marrow Mesenchymal Stromal Cells in Patients With Aplastic Anemia: A Systematic Review. Front Immunol. 2022;13:859668. https://doi.org/10.3389/fimmu.2022.859668

Daull P, Barabino S, Feraille L, Kessal K, Docquier M, Parsadaniantz S M, et al. Modulation ‎of inflammation-related genes in the cornea of ‎a mouse model of dry eye upon treatment with ‎cyclosporine eye drops. Curr. Eye Res. 2019;44(5):476-485. 10.1080/02713683.2018.1563197

Sun HJ, Song YS, Cho SW, Park YJ. Enhancement of Osteogenic Differentiation by ‎Combination Treatment with 5-azacytidine and Thyroid-‎Stimulating Hormone in Human ‎Osteoblast Cells. Int. J. Thyroidol. 2017;10(2):71-76. https://doi.org/10.11106/ijt.2017.10.2.71

Araak JK. The protective role of date palm pollen (Phoenix dactylifera L.) on liver function ‎in adult male rats treated with carbon ‎tetrachloride. Iraqi J. Vet. Med. 2012;36(0E):132-142.


Zhou GS, Zhang XL, Wu JP, Zhang RP, Xiang LX, Dai LC, Shao JZ. 5-Azacytidine facilitates osteogenic gene expression and differentiation of mesenchymal stem cells by alteration in DNA methylation. Cytotechnology. 2009;60(1-3):11-22. https://doi.org/10.1007/s10616-009-9203-2

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