Ratlarda Siklofosfamid ile Deneysel Olarak Oluşturulan Anemi Modelinde Smilax excelsa L. Etanol Ekstresinin Etkisi

Abstract

Bu çalışmada, ratlarda siklofosfamid (CP) ile deneysel olarak oluşturulan anemi modelinde Smilax excelsa L. etanol ekstresinin etkisi araştırıldı. Çalışma toplam 4 gruptan oluştu. Grup 1 ve 2’deki ratlara oral gavaj yöntemi kullanılarak 28 gün süre ile günlük 1 ml serum fizyolojik, grup 3 ve 4’deki ratlara ise 400 mg/kg dozda Smilax excelsa L. etanol ekstresi uygulandı. Ayrıca grup 2 ve 4’deki ratlara haftada bir kez olmak üzere toplam dört doz 50 mg/kg CP uygulaması intramusküler olarak yapıldı. Çalışmanın 28. günü bütün ratlardan anestezi altında kan örnekleri alındı ve daha sonra ötanazi işlemi uygulandı. Kan örneklerinde eritrosit sayısı (RBC), lökosit sayısı (WBC), hemoglobin konsantrasyonu (HGB), hematokrit değer (Hct), ortalama eritrosit hemoglobini (MCH), ortalama eritrosit hacmi (MCV) ve ortalama eritrosit dağılım genişliği (RDW) gibi hematolojik parametreler incelendi. Serum malondialdehit (MDA) ve indirgenmiş glutatyon (GSH) düzeyleri ile katalaz (CAT) ve glutatyonperoksidaz (GSH-Px) enzim aktivitelerine spektrofotometrik olarak bakıldı. Grup 4’de Smilax excelsa L. etanol ekstresi tedavisinin serum MDA düzeyini düşürdüğü, CAT ve GSH-Px enzim aktivitelerini ise arttırdığı belirlendi. Ayrıca bu tedavinin RBC, HGB ve Hct gibi hematolojik parametrelerde iyileşmelere sebep olduğu, RDW değerlerini ise azalttığı gözlemlendi. Sonuç olarak Smilax excelsa’nın sahip olduğu güçlü antioksidan etki ile CP’nin kemik iliğindeki baskılayıcı etkisini azalttığı ve anemi şekillenmesini önlediği görüldü.

Keywords

• Aplastik Anemi
• Siklofosfamid
• Oksidatif Stres
• Smilasagiller

The Effect of Smilax excelsa L. Ethanol Extract in an Experimentally Anemia Model İnduced by Cyclophosphamide in Rats

Abstract

In this study, the effect of Smilax excelsa L. ethanol extract on anemia model experimentally induced by Cyclophosphamide (CP) in rats was investigated. The study consisted of 4 groups, The rats in groups 1 and 2 were administered 1 ml of physiological saline for 28 days using the oral gavage method, and the rats in groups 3 and 4 were administered Smilax excelsa L. ethanol extract at a dose of 400 mg/kg. In addition, four doses of 50 mg/kg CP were administered intramuscularly once a week to the rats in groups 2 and 4. On the 28th day of the experiment, blood samples were taken from all rats under anesthesia and then euthanasia was performed. Hematological parameters such as red blood cell count (RBC), leukocyte count (WBC), hemoglobin concentration (HGB), hematocrit value (Hct), mean erythrocyte hemoglobin (MCH), mean erythrocyte volume (MCV), and mean erythrocyte distribution width (RDW) were examined in blood samples. Serum malondialdehyde (MDA) and reduced glutathione (GSH) levels and catalase (CAT) and glutathione peroxidase (GSH-Px) enzyme activities were measured spectrophotometrically. In group 4, it was determined that Smilax excelsa L. ethanol extract treatment decreased the serum MDA level and increased the CAT and GSH-Px enzyme activities. In addition, it was observed that this treatment caused improvements in hematological parameters such as RBC, HGB and Hct, and decreased RDW values. As a result, it was observed that Smilax excelsa, with its strong antioxidant effect, reduced the suppressive effect of CP in the bone marrow and prevented the formation of anemia.

Keywords

• Aplastic Anemia
• Cyclophosphamide
• Oxidative Stress
• Smilacaceae

References

1. Aebi H (1983). Catalase. In H. U. Bergmeyer (Ed.), Methods of enzymatic analysis. Verlag Chemie. pp. 273-286, Weinhem, Germany.
2. Anokwuru CP, Anyasor GN, Shokunbi OS et al. (2019). Chemoprotective activity of aqueous leaf extract of Acalypha wilkesiana against cyclophosphamide-induced toxicity in rats. Asian Pac J Trop Med, 12 (9), 409-415.
3. Anyasor GN, Oyewole IO, Ogunwenmo KO, Ayomide A. (2012). Coartemether induced oxidative and hepatic damage in Plasmodium berghei strain Anka infected mice. Bull Environ Contam Toxicol, 1, 108-111.
4. Ashry NA, Gameil NM, Suddek GM (2013). Modulation of cyclophosphamide- induced early lung injury by allicin. Pharm Biol, 51, 806-811.
5. Ayhancı A, Heybeli N, Kulcanay Şahin İ, Cengiz M (2019). Myelosuppression and oxidative stress induced by cyclophosphamide in rats: The protective role of selenium. Adıyaman Univ J of Sci, 9 (2), 252-265.
6. Bai Y, Jiang Y, Liu T et al. (2018). Xinjiang herbal tea exerts immunomodulatory activity via TLR2/4-mediated MAPK signaling pathways in RAW264.7 cells and prevents cyclophosphamideinduced immunosuppression in mice. J Ethnopharmacol, 228,179-187.
7. Brigle K, Pierre A, Finley-Oliver E et al. (2017). Myelosuppression, bone disease, and acute renal failure: evidence-based recommendations for oncologic emergencies. Clin J Oncol Nurs, 21 (5 Suppl. l), 60-76.
8. Chen YF, Zhao ZQ, Wu ZM et al. (2014). The role of RIP1 and RIP3 in the development of aplastic anemia induced by cyclophosphamide and busulphan in mice. Int J Clin Exp Pathol, 7 (12), 8411-20.

9. Chukwuemeka I, Utuk GS, Ugwu Okechukwu PC et al. (2015). The effect of ethanol leaf extract of Jatropha curcas on some haematological parameters of cyclophosphomide induced anaemia in wister albino rats. EJAS, 7 (1), 17-20.
10. Dame C, Kirschner KM, Bartz KV et al. (2006). Wilms tumor suppressor, Wt1, is a transcriptional activator of the erythropoietin gene. Blood, 107, 4282-4290.
11. Elhalim SAA, Sharada HM, Abulyazid I, Aboulthana WM, Elhalim STA (2017). Ameliorative effect of carob pods extract (Ceratonia siliqua L.) against cyclophosphamide induced alterations in bone marrow and spleen of rats. J Appl Pharm Sci, 7 (10), 168-181.
12. El-Sebaey AM, Abdelhamid FM, Abdalla OA (2019). 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.
13. Eltantawy FM, Sobh MAA, EL-Waseef AM, Ibrahim RAA, Saad MAA (2018). Protective effect of Spirulina against cyclophosphamide-induced urotoxicity in mice. Egypt j basic appl sci, 5, 191-196.
14. Goudarzi M, Khodayar MJ, Mohammad S et al. (2017). Pretreatment with melatonin protects against cyclophosphamide-induced oxidative stress and renal damage in mice. Fundam Clin Pharmacol, 31, 625-635.
15. Han J, Dai M, Zhao Y et al. (2020). Compatibility effects of ginseng and Ligustrum lucidum Ait herb pair on hematopoietic recovery in mice with cyclophosphamide-induced myelosuppression and its material basis. J Ginseng Res, 44, 291-299.
16. He M, Wang N, Zheng W et al. (2021). Ameliorative effects of ginsenosides on myelosuppression induced by chemotherapy or radiotherapy. J Ethnopharmacol, 268, 113581.
17. Ikumawoyi VO, Awodele O, Rotimi K, Fashina AY (2016). Evaluation of The Effects of the hydro-ethanolic root extract of Zanthoxylum zanthoxyloides on hematological parameters and oxiative stress in cyclophospamide treated rats. Afr J Tradit Complement Altern Med, 13 (5), 153-159.
18. Iqubal A, Syed MA, Haque MM et al. (2020). Effect of nerolidol on cyclophosphamide-induced bone marrow and hematologic toxicity in swiss albino mice. Exp Hematol, 82, 24-32.
19. Jia W, Zhen M, Li L et al. (2020). Gadofullerene nanoparticles for robust treatment of aplastic anemia induced by chemotherapy drugs. Theranostics, 10 (15), 6886-6897.
20. Kalantar M, Goudarzi M, Khodayar MJ et al. (2016). Protective effect of the hydroalcoholic extract of Capparis spinose L. against cyclophosphamide-induced nephrotoxicity in mice. Jundishapur J Nat Pharm Prod, 11 (4), 37240.
21. Khan JA, Shahdad S, Makhdoomi MA et al. (2014). Effect of cyclophosphamide on the microanatomy of liver of albino rats. Int J Res Med Sci, 2 (4), 1466-1469.
22. Khorwal G, Chauhan R, Nagar M (2017). Effect of cyclophosphamide on liver in albino rats: A comparative dose dependent histomorphological study. Int J Biomed Adv Res, 8 (3), 102-107.
23. Kim JW, Choi JS, Seol DJ, Choung JJ, Kwang Ku SK (2018). Immunomodulatory effects of Kuseonwangdogo-based mixed herbal formula extracts on a cyclophosphamide-induced immunosuppression mouse model. eCAM, Article ID 6017412, 14 .
24. Kulshrestha MK, Mahapatra KB, Das AK (2019). Haematopoietic activity of Mandoor parpati on cyclophosphamide induced anaemia in wistar strain albino rats. Bull Env Pharmacol Life Sci, 8 (3), 98-102.
25. Lawrence RA, Raymond FB (1976). Glutathione peroxidase activity in selenium deficient rat liver. Biochem Biophys Res, 71 (4), 952-958.
26. Li F, Tang R, Chen LB et al. (2017). Effects of Astragalus combined with angelica on bone marrow hematopoiesis suppression induced by cyclophosphamide in mice. Biol Pharm Bull, 40, 598-609.
27. Lowry OH, Rosebrough NJ, Farr AL, Randall RJ (1951). Protein measurement with the Folin phenol reagent. J Biol Chem, 193, 265-275.
28. Murthy HN, Park SY, Lee EJ, Paek KY (2015). Biosafety and toxicological evaluation of tissue-cultured Echinacea purpurea adventitious roots. Korean J Hortic Sci Technol, 33 (1), 124-132.
29. Mythili Y, Sudharsan PT, Selvakumar E, Varalakshmi P (2004). Protective effect of DL-alpha-lipoic acid on cyclophosphamide induced oxidative cardiac injury. Chem Biol Interact, 151 (1), 13-19.
30. Özsoy N, Okyar A, Arda Pirinçci P et al. (2016). The Protective effect of an aqueous extract from Smilax excelsa L. against carbon tetrachloride-induced liver in rats. Afr J Tradit Complement Altern Med, 13 (3), 203-208.
31. Özsoy N, Okyar A, Arda Pirinçci P et al. (2013). Evaluation of Smilax excelsa L. use in experimentally induced nephrotoxicity. Kafkas Univ Vet Fak Derg, 19 (5), 807-814.
32. Özsoy N, Can A, Yanardag R, Akev N (2008). Antioxidant activity of Smilax excelsa L. leaf extracts. Food Chem, 110 (3), 571-583.
33. Perini P, Calabrese M, Rinaldi L, Gallo P (2007). The safety profile of cyclophosphamide in multiple sclerosis therapy. Expert Opin Drug Safety, 6, 183-90.
34. Placer ZA, Cushman LL, Johnson BC (1966). Estimation of product of lipid peroxidation (malonyldialdehyde) in biochemical systems. Anal Biochem, 16 (2), 359-364
35. Raj SV (2009). Management of chemotherapy-induced thrombocytopenia: current status of thrombopoietic agents. Semin Hematol, 46 (2), 26-32.
36. Ramadan G, El-Beih NM, Zahra MM (2012). Egyptian sweet marjoram leaves protect against genotoxicity, immunosuppression and other complications induced by cyclophosphamide in albino rats. Br J Nutr, 108, 1059-1068.
37. Rostampur S, Feizi MAH, Khojasteh SMB, Daluchi F (2018). Heracleum persicum extract improves cyclophosphamide-induced liver toxicity and oxidative stress in male rats. Adv Herb Med, 4 (3), 34-44.
38. Sedlak J, Lindsay RH (1968). Estimation of total, protein-bound, and nonprotein sulfhydryl groups in tissue with Ellman’s reagent. Anal Biochem, 25, 192-205.
39. Süzek H, Çelik I, Doğan A (2017). Nephroprotective hepatoprotective potential and antioxidant role of carob pods (Cerotonia siliqua L.) against carbon tetrachloride-induced toxicity in rats. Indian J Pharm Educ Res, 51 (2), 312-320.
40. Wang Z, Qi F, Cui Y et al. (2018). An update on Chinese herbal medicines as adjuvant treatment of anticancer therapeutics. Biosci Trends, 12, 220-239.
41. Woo S, Krzyzanski W, Jusko WJ (2008). Pharmacodynamic model for chemotherapy-induced anemia in rats. Cancer Chemother Pharmacol, 62, 123-133.
42. Xu M, He RR, Zhai YJ, Abe K, Kurihara H (2014). Effects of carnosine on cyclophosphamide-induced hematinic suppression in mice. Am J Chin Med, 42, 131-142.
43. Yıldız ÖŞ, Ayanoğlu F, Bahadırlı NP (2018). Saparna (Smilax aspera L., Smilax excelsa L.) Türlerinin Bazı Morfolojik ve Kimyasal Özellikleri. MKÜ Ziraat Fakültesi Dergisi, 23 (2), 254-261.
44. Zhang J, Tian Q, Zhou S (2006). Clinical pharmacology of cyclophosphamide and ifosfamide. Curr Drug Ther, 1, 55-84.
45. Zhang L, Gong AGW, Riaz K et al. (2017). A novel combination of four flavonoids derived from Astragali Radix relieves the symptoms of cyclophosphamide-induced anemic rats. FEBS Open Bio, 7, 318-32