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Protective effects of garlic extract against hematological alterations, immunosuppression, hepatic oxidative stress, and renal damage induced by cyclophosphamide in rats

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Abstract

Cyclophosphamide is an alkylating agent widely used as anticancer drug, reported to exert cytotoxic effects attributed to oxidative stress. Therefore, this study aimed to explore the protective effect of ethanolic extract of garlic (EEG) against cyclophosphamide (Cyp)-induced hematological disturbance and immunosuppressive and hepatotoxic effects. Forty male Wistar albino rats were randomized into four equal groups: the normal control one, the Cyp-treated group (50 mg/kg BW/IM, once weekly), the EEG-treated group (300 mg/kg BW, orally, daily), and the Cyp & EEG group. All rats received their relevant treatments for four consecutive weeks. This study revealed that Cyp significantly decreased erythrocyte count, hemoglobin (Hb), packed cell volume (PCV), and total leukocyte and lymphocyte counts. However, the counts of neutrophils, eosinophils, and toxic neutrophils were elevated. Additionally, hepatic malondialdehyde (MDA) and levels of liver and renal biomarkers were significantly elevated in the Cyp-treated group. Otherwise, hepatic catalase (CAT), reduced glutathione (GSH), superoxide dismutase (SOD), and serum total antioxidant capacity (TAC) were significantly lower than the control rats. Furthermore, Cyp significantly reduced whole blood respiratory burst activity (NBT), serum lysozyme and bactericidal activities, interlukin-12 (IL-12), and interferon-γ. In contrast, the levels of nitric oxide (NO), tumor necrosis factor-α (TNF-α), and interlukin-1β (IL-1β) were elevated. Additionally, Cyp induced hepatic and renal histopathological alterations. Data in the present study demonstrated that EEG has immunomodulatory and antioxidant effects and has the ability to diminish the alterations induced by Cyp.

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References

  • Abdul Qadir M, Shahzadi SK, Bashir A, Munir A, Shahzad S (2017) Evaluation of phenolic compounds and antioxidant and antimicrobial activities of some common herbs. Int J Anal Chem 6:1–6. https://doi.org/10.1155/2017/3475738

    Article  CAS  Google Scholar 

  • Akinbi HT, Epaud R, Bhatt H, Weaver TE (2000) Bacterial killing is enhanced by expression of lysozyme in the lungs of transgenic mice. J Immunol 165:5760–5766. https://doi.org/10.4049/jimmunol.165.10.5760

    Article  CAS  Google Scholar 

  • Alabbassi MG (2010) Melatonin ameliorates hepatic damage induced by cyclophosphamide in rats. J Pharm Sci 17:47–54. https://doi.org/10.1111/and.12191

    Article  CAS  Google Scholar 

  • Al-Malki AL (2014) Synergestic effect of lycopene and melatonin against the genesis of oxidative stress induced by cyclophosphamide in rats. Toxicol Ind Health 30:570–575. https://doi.org/10.1177/0748233712459916

    Article  CAS  Google Scholar 

  • Anusuya N, Durgadevi P, Dhinek A, Mythily S (2013) Nephroprotective effect of ethanolic extract of garlic (Allium sativum) on cisplatin induced nephrotoxicity in male Wistar rats. Asian J Pharm Clin Res 6:97–100

    Google Scholar 

  • Asdaq SMB (2015) Antioxidant and hypolipidemic potential of aged garlic extract and its constituent, S-allyl cysteine, in rats. Evid Based Complement Alternat Med 7:1–7. https://doi.org/10.1155/2015/328545

    Article  Google Scholar 

  • Bancroff JP, Stevenes A, Turner DR (1990) Theory and practice of histological techniques, 3rd edn. Clurechill Livingston, Edinburgh, London

    Google Scholar 

  • Berger J, Slapničková M (2003) Circadian structure of rat neutrophil phagocytosis. Comp Clin Path 12:84–89. https://doi.org/10.1007/s00580-003-0478-y

    Article  Google Scholar 

  • Boonpeng S, Siripongvutikorn S, Sae-wong C, Sutthirak P (2014) The antioxidant and anti-cadmium toxicity properties of garlic extracts. F Sci Nut 2:792–801. https://doi.org/10.1002/fsn3.164

    Article  CAS  Google Scholar 

  • Casanova NA, Simoniello MF, López Nigro MM, Carballo MA (2017) Modulator effect of watercress against cyclophosphamide-induced oxidative stress in mice. Medicina (BA) 77:201–206

    Google Scholar 

  • Chang CC, Yang MH, Wen HM, Chern JC (2002) Estimation of total flavonoid content in propolis by two complementary colorimetric methods. J Food Drug Anal 10:178–182

    CAS  Google Scholar 

  • Dame C, Kirschner KM, Bartz KV, Wallach T, Hussels CS, Scholz H (2006) Wilms tumor suppressor, Wt1, is a transcriptional activator of the erythropoietin gene. Blood 107:4282–4290

    Article  CAS  Google Scholar 

  • Dong Q, Sugiura T, Toyohira Y, Yoshida Y, Yanagihara N, Karasaki Y (2011) Stimulation of IFN-γ production by garlic lectin in mouse spleen cells: involvement of IL-12 via activation of p38 MAPK and ERK in macrophages. Phytomedicine 18:309–316

    Article  CAS  Google Scholar 

  • Duggina P, Kalla CM, Varikasuvu SR, Bukke S, Tartte V (2015) Protective effect of centella triterpene saponins against cyclophosphamide-induced immune and hepatic system dysfunction in rats: its possible mechanisms of action. J Physiol Biochem 71:435–454. https://doi.org/10.1007/s13105-015-0423-y

    Article  CAS  Google Scholar 

  • El-Banna SG, Attia AM, Hafez AM, El-Kazaz SM (2009) Effect of garlic consumption on blood lipid and oxidant-antioxidant parameters in rat males exposed to chlorpyrifos. Slolvak J of Animal Science (Slov Rep) 42:111–117

    Google Scholar 

  • El-Olemy MM, Al-Muhtadi FJ, Afifi AFA (1994) Experimental phytochemistry: a laboratory manual. Department of Pharmacognosy, College of Pharmacy, King Saud University Press, Saudi Arabia, pp 21–27

    Google Scholar 

  • Fahmy SR, Amien AI, Abd-Elgleel FM, Elaskalany SM (2016) Antihepatotoxic efficacy of Mangifera indica L. polysaccharides against cyclophosphamide in rats. Chem Biol Interact 244:113–120. https://doi.org/10.1016/j.cbi.2015.11.009

    Article  CAS  Google Scholar 

  • Fandrey J, Frede S, Jelkmann W (1994) Role of hydrogen peroxide in hypoxia-induced erythropoietin production. Biochem J 303:507–510

    Article  CAS  Google Scholar 

  • Feldman BF, Zinkl JG, Jain VC (2000) Schalm’s veterinary hematology, 5th edn. Lippincott Williams and Wilkins, Toronto, Canada, pp 1145–1146

    Google Scholar 

  • Förstermann U, Schmidt HH, Pollock JS, Sheng H, Mitchell JA, Warner TD, Nakane M, Murad F (1991) Isoforms of nitric oxide synthase characterization and purification from different cell types. Biochem Pharmacol 42:1849–1857. https://doi.org/10.1016/0006-2952(91)90581-O

    Article  Google Scholar 

  • Fotakis G, Timbrell JA (2006) In vitro cytotoxicity assays: comparison of LDH, neutral red, MTT and protein assay in hepatoma cell lines following exposure to cadmium chloride. Toxicol Lett 160:171–177. https://doi.org/10.1016/j.toxlet.2005.07.001

    Article  CAS  Google Scholar 

  • Germoush MO, Mahmoud AM (2014) Berberine mitigates cyclophosphamide-induced hepatotoxicity by modulating antioxidant status and inflammatory cytokines. J Cancer Res Clin Oncol 140:1103–1109. https://doi.org/10.1007/s00432-014-1665-8

    Article  CAS  Google Scholar 

  • Goyal S, Gupta N, Chatterjee S (2016) Investigating therapeutic potential of Trigonella foenum-graecum L. as our defense mechanism against several human diseases. J Toxicol 2:1–10. https://doi.org/10.1155/2016/1250387

    Article  CAS  Google Scholar 

  • Hamlaoui-Gasmi S, Mokni M, Limam N, Limam F, Amri M, Aouani E, Marzouki L (2014) Effect of garlic’s mode of administration on erythrocytes and plasma parameters in Wistar rat. Afr J Biotechnol 11:8259–8263

    Google Scholar 

  • Hickman-Davis JM, Lindsey JR, Matalon S (2001) Cyclophosphamide decreases nitrotyrosine formation and inhibits nitric oxide production by alveolar macrophages in mycoplasmosis. Infect Immun 69:6401–6410. https://doi.org/10.1128/IAI.69.10.6401-6410.2001

    Article  CAS  Google Scholar 

  • Higuchi T, Bodin JI (1961) In: Higuchi T, Hansen EB (eds) Alkaloid and other basic nitrogenous compounds; in pharmaceutical analysis (edns.). Interscience, New York, pp 315–345

    Google Scholar 

  • Huang JQ, Pang MR, Li GY, Wang N, Jin L, Zhang Y (2017) Alleviation of cyclophosphamide-induced immunosuppression in mice by naturally acetylated hemicellulose from bamboo shavings. Food Agric Immunol 28:328–342. https://doi.org/10.1080/09540105.2016.1272553

    Article  CAS  Google Scholar 

  • Inoue LAKA, Oliveira Maciel P, Gusmão Affonso E, Lima Boijink C, Tavares-Dias M (2016) Growth, parasitic infection and hematology in Colossoma macropomum Cuvier, 1818 fed diets containing Allium sativum. J Appl Ichthyol 32:901–905. https://doi.org/10.1111/jai.13086

    Article  CAS  Google Scholar 

  • Khalil R, El-Hofy H, Yehya MM, Selim KM (2014) Some biochemical and immunological changes associated with use of garlic extract (allicin) in combating some fish pathogens. Alex J Vet Sci 41:1–18. https://doi.org/10.5455/ajvs.152049

    Article  Google Scholar 

  • Khan MA (2017) Immune potentiating and antitoxic effects of camel milk against cyclophosphamide-induced toxicity in BALB/C mice. Int J Health Sci 11:1–18

    Google Scholar 

  • Kilikdar D, Mukherjee D, Mitra E, Ghosh AK, Basu A, Chandra AM, Bandyoapdhyay D (2011) Protective effect of aqueous garlic extract against lead-induced hepatic injury in rats. Indian J Exp Biol 49:498–510

    Google Scholar 

  • Kim SH, Lee IC, Ko JW, Moon C, Kim SH, Shin IS, Kim JC (2015) Diallyl disulfide prevents cyclophosphamide-induced hemorrhagic cystitis in rats through the inhibition of oxidative damage, MAPKs, and NF-κB pathways. Biomol Ther (Seoul) 23:180–188. https://doi.org/10.4062/biomolther.2014.126

    Article  CAS  Google Scholar 

  • Klaassen CD, Casarett LJ, Doull J (2008) Casarett and Doull’s toxicology: the basic science of poisons, 7th edn. McGraw-Hill, New York

    Google Scholar 

  • Kocahan S, Dogan Z, Erdemli E, Taskin E (2017) Protective effect of quercetin against oxidative stress induced toxicity associated with doxorubicin and cyclophosphamide in rat kidney and liver tissue. Iran J Kidney Dis 11:124–131

    Google Scholar 

  • Kokate CK, Purohit AP, Gokhale SB (1995) Pharmacognosy, 3rd edn. Niralin Prakashan, Pune, p 125

    Google Scholar 

  • Kumar VP, Venkatesh YP (2016) Alleviation of cyclophosphamide-induced immunosuppression in Wistar rats by onion lectin (Allium cepa agglutinin). J Ethnopharmacol 186:280–288. https://doi.org/10.1016/j.jep.2016.04.006

    Article  CAS  Google Scholar 

  • Lindsay H (1973) A colorometric estimation of reducing sugar in potatoes with 3-5 dinitrosalicylic acid. Potatoes Res 16:176–179. https://doi.org/10.1007/BF02356048

    Article  CAS  Google Scholar 

  • Madondo MT, Quinn M, Plebanski M (2016) Low dose cyclophosphamide: mechanisms of T cell modulation. Cancer Treat Rev 42:3–9. https://doi.org/10.1016/j.ctrv.2015.11.005

    Article  Google Scholar 

  • Mahmoud AM (2014) Hesperidin protects against cyclophosphamide-induced hepatotoxicity by upregulation of PPARγ and abrogation of oxidative stress and inflammation. Can J Physiol Pharmaco 92:717–724. https://doi.org/10.1139/cjpp-2014-0204

    Article  CAS  Google Scholar 

  • Mahmoud AM, Wilkinson FL, Jones AM, Wilkinson JA, Romero M, Duarte J, Alexander MY (2017) A novel role for small molecule glycomimetics in the protection against lipid-induced endothelial dysfunction: involvement of Akt/eNOS and Nrf2/ARE signaling. Biochim Biophys Acta Gen Subj 1861:3311–3322. https://doi.org/10.1016/j.bbagen.2016.08.013

    Article  CAS  Google Scholar 

  • Merwid-Ląd A, Ksiądzyna D, Hałoń A, Chlebda-Sieragowska E, Trocha M, Szandruk M, Sozański T, Magdalan J, Kopacz M, Kuźniar A, Nowak D, Pieśniewska M, Szeląg A (2015) Impact of morin-5′-sulfonic acid sodium salt on cyclophosphamide-induced gastrointestinal toxicity in rats. Pharmacol Rep 67:1259–1263. https://doi.org/10.1016/j.pharep.2015.05.020

    Article  CAS  Google Scholar 

  • Mikaili P, Maadirad S, Moloudizargari M, Aghajanshakeri S, Sarahroodi S (2013) Therapeutic uses and pharmacological properties of garlic, shallot, and their biologically active compounds. Iran J Basic Med Sci 16:1031–1048

    CAS  Google Scholar 

  • Mirfardi M, Johari H (2015) The effect of hydro-alcoholic Allium sativum extract on sexual hormones in mature male rats under chemotherapy with cyclophosphamide. Zahedan J Res Med Sci 17:29–33

    Article  CAS  Google Scholar 

  • Mohammed NA, Abd El-Aleem SA, El-Hafiz HA, McMahon RF (2004) Distribution of constitutive (COX-1) and inducible (COX-2) cyclooxygenase in postviral human liver cirrhosis: a possible role for COX-2 in the pathogenesis of liver cirrhosis. J Clin Pathol 57:350–354

    Article  CAS  Google Scholar 

  • Montgomery HAC, Dymock JF (1961) Determination of nitric oxide in water. Analyst (London) 86:414–416

    CAS  Google Scholar 

  • Motawi TM, Sadik NA, Refaat A (2010) Cytoprotective effects of DL-alpha-lipoic acid or squalene on cyclophosphamide-induced oxidative injury: an experimental study on rat myocardium, testicles and urinary bladder. Food Chem Toxicol 48:2326–2336. https://doi.org/10.1016/j.fct.2010.05.067

    Article  CAS  Google Scholar 

  • Nasr AY (2014) Protective effect of aged garlic extract against the oxidative stress induced by cisplatin on blood cells parameters and hepatic antioxidant enzymes in rats. Toxi Rep 1:682–691. https://doi.org/10.1016/j.toxrep.2014.09.003

    Article  CAS  Google Scholar 

  • Ndong D, Fall J (2011) The effect of garlic (Allium sativum) on growth and immune responses of hybrid tilapia (Oreochromis niloticus x Oreochromis aureus). J Clin Immunol Immunopathol Res 3:1–9

    Google Scholar 

  • Neboh EE, Ufelle SA (2015) Myeloprotective activity of crude methanolic leaf extract of Cassia occidentalis in cyclophosphamide-induced bone marrow suppression in Wistar rats. Adv Biomed Res 4:5. https://doi.org/10.4103/2277-9175.148285

    Article  Google Scholar 

  • Niki E, Noguchi N, Tsuchihashi H, Gotoh N (1995) Interaction among vitamin C, vitamin E, and beta-carotene. Am J Clin Nutr 62:1322S–1326S

    Article  CAS  Google Scholar 

  • Olaniyan OT, Meraiyebu AB, Anjorin YD, Shekins O, Dare BJ, Shafe MO (2013) Effects of aqueous extract of onion (Allium sativum) on blood parameters in adult wistar rats (Rattus novergicus). IJPSI 2:42–45

    Google Scholar 

  • Ozougwu JC (2011) An investigation of the effects of Allium Sativum (garlic) extracts on haematological profile of white albino rats. Pharmacologonline 2:299–306

    Article  CAS  Google Scholar 

  • Padiya R, Khatua TN, Bagul PK, Kuncha M, Banerjee SK (2011) Garlic improves insulin sensitivity and associated metabolic syndromes in fructose fed rats. Nut Metab (Lond) 8:53. https://doi.org/10.1186/1743-7075-8-53

    Article  CAS  Google Scholar 

  • Park KJ, Lee BC, Lee JS, Cho MH (2014) Angelica gigas Nakai extract ameliorates the effects of cyclophosphamide on immunological and hematopoietic dysfunction in mice. J Med Plant Res 8:657–663. https://doi.org/10.5897/JMPR2013.5288

    Article  CAS  Google Scholar 

  • Peng JP, Yao X, Kobayashi H, Ma C (1995) Novel furastonol glycosides from Allium macrostemon. Planta Med 6:58–61. https://doi.org/10.1055/s-2006-958000

    Article  Google Scholar 

  • Price ML, Butter LG (1977) Rapid visual estimation and spectrophotometric determination of tanning content of sorghum grain. J Agric Food Chem 25:1268–1273

    Article  CAS  Google Scholar 

  • Rubinstein MP, Su EW, Suriano S, Cloud CA, Andrijauskaite K, Kesarwani P, Schwartz KM, Williams KM, Johnson CB, Li M, Scurti GM, Salem ML, Paulos CM, Garrett-Mayer E, Mehrotra S, Cole DJ (2015) Interleukin-12 enhances the function and anti-tumor activity in murine and human CD8+ T cells. Cancer Immunol Immunother 64:539–549. https://doi.org/10.1007/s00262-015-1655-y

    Article  CAS  Google Scholar 

  • Sahoo PK, Mukherjee SC (2001) Immunosuppressive effects of aflatoxin B1 in Indian major carp (Labeo rohita). Comp Immunol Microbiol Infect Dis 2:143–149. https://doi.org/10.1016/S0147-9571(00)00017-5

    Article  Google Scholar 

  • Sasaki G, Satoh T, Yokozeki H, Katayama I, Nishioka K (2000) Regulation of cyclophosphamide-induced eosinophilia in contact sensitivity: functional roles of interleukin-5-producing CD4+ lymphocytes. Cell Immunol 203:124–133. https://doi.org/10.1006/cimm.2000.1677

    Article  CAS  Google Scholar 

  • Seckiner I, Bayrak O, Can M, Mungan AG, Mungan NA (2014) Garlic supplemented diet attenuates gentamicin nephrotoxicity ın rats. Int Braz J Urol 40:562–567. https://doi.org/10.1590/S1677-5538.IBJU.2014.04.17

    Article  Google Scholar 

  • Selvakumar E, Prahalathan C, Mythili Y, Varalakshmi P (2005) Mitigation of oxidative stress in cyclophosphamide-challenged hepatic tissue by DL-α-lipoic acid. Mol Cell Biochem 272:179–185

    Article  CAS  Google Scholar 

  • Senthilkumar S, Devaki T, Manohar BM, Babu MS (2006) Effect of squalene on cyclophosphamide-induced toxicity. Clin Chim Acta 364:335–342. https://doi.org/10.1016/j.cca.2005.07.032

    Article  CAS  Google Scholar 

  • Singleton VL, Rossi JA (1965) Colorimetry of total phenolics with phosphomolybdic-phosphotungstic acid reagents. Am J Enol Vitic 16:144–158

    CAS  Google Scholar 

  • Song Y, Zhang C, Wang C, Zhao L, Wang Z, Dai Z, Ma X (2016) Ferulic acid against cyclophosphamide-induced heart toxicity in mice by inhibiting NF-κB pathway. Evid Based Complement Alternat Med 8:1–8. https://doi.org/10.1155/2016/1261270

    Article  CAS  Google Scholar 

  • Sowjanya BL, Devi KR, Madhavi D (2009) Modulatory effects of garlic extract against the cyclophosphamide induced genotoxicity in human lymphocytes in vitro. J Environ Biol 30:663–666

    CAS  Google Scholar 

  • Swamy AV, Patel UM, Koti BC, Gadad PC, Patel NL, Thippeswamy AHM (2013) Cardioprotective effect of Saraca indica against cyclophosphamide induced cardiotoxicity in rats: a biochemical, electrocardiographic and histopathological study. Indian J Pharmacol 45:44–48. https://doi.org/10.4103/0253-7613.106434

    Article  Google Scholar 

  • Wani AA, Sikdar-Bar M (2014) Ameliorative efficacy of taurine and garlic extract on copper induced immunotoxic effect on total and differential leucocyte counts in African catfish, Clarias gariepinus. Asian J Med Pharm Res 4:122–129

    Google Scholar 

  • Wójcik R (2014) Reactivity of the immunological system of rats stimulated with Biolex-Beta HP after cyclophosphamide immunosuppression. Cent Eur J Immunol 39:51–60. https://doi.org/10.5114/ceji.2014.42125

    Article  CAS  Google Scholar 

  • Wojcik R, Dabkowska A (2010) The effect of cyclophosphamide on the selected parameters of immunity in rats. Cent Eur J Immunol 35:1–9

    CAS  Google Scholar 

  • Zarei M, Shivanandappa T (2013) Amelioration of cyclophosphamide-induced hepatotoxicity by the root extract of Decalepis hamiltonii in mice. Food Chem Toxicol 57:179–184. https://doi.org/10.1016/j.fct.2013.03.028

    Article  CAS  Google Scholar 

  • Zhao R, Ma C, Tan L, Zhao X, Zhuang D (1994) The effect of acupuncture on the function of macrophages in rats of immunodepression. Zhen Ci Yan Jiu 19:66–68

    CAS  Google Scholar 

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Acknowledgments

The authors are deeply grateful to Professor Dr. Mohamed El-Boshy, Laboratory Medicine Department, Faculty of Applied Medical Science, Umm Al-Qura University, PB 7296, Makkah 21955, Saudi Arabia, for his kind help in statistical analysis.

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Authors and Affiliations

  1. Clinical Pathology Department, Faculty of Veterinary Medicine, Mansoura University, Mansoura, 35516, Egypt

    Ahmed M. El-Sebaey & Fatma M. Abdelhamid

  2. Clinical Pathology Department, Faculty of Veterinary Medicine, Suez Canal University, Ismailia, Egypt

    Osama A. Abdalla

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  1. Ahmed M. El-Sebaey
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Correspondence to Fatma M. Abdelhamid.

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El-Sebaey, A.M., Abdelhamid, F.M. & Abdalla, O.A. Protective effects of garlic extract against hematological alterations, immunosuppression, hepatic oxidative stress, and renal damage induced by cyclophosphamide in rats. Environ Sci Pollut Res 26, 15559–15572 (2019). https://doi.org/10.1007/s11356-019-04993-7

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