Evaluation of the combined effects of Turkish mad honey and 5-Fluorouracil in colon cancer model in rats

Abstract

It was aimed to evaluate the regressive effect of grayanotoxin-rich Turkish mad honey and 5-fluorouracil (5-FU), separately and together by using the N-methyl-N-nitrosourea (MNU)-induced colon cancer modelling in rats. Study groups were designed as control group (CG), cancer control group (CCG), 5-Flourouracil group (FUG), Turkish mad honey group (HG), Turkish mad honey and 5-FU combined group (FU-HG). White blood cell (WBC), lymphocyte, eosinophil, basophil, serum lactate dehydrogenase (LDH), total oxidant status (TOS), and total protein values of the rats in the CCG were significantly lower than the values of the rats in the CG, whereas serum Bcl-2 and survivin levels were significantly higher in the rates belonged to the CCG in comparison to those in the CG. The presence of anaplastic epithelial cells, vascularization, precancerous changes, and inflammatory infiltration detected in the colon and small intestine of the rats in FU-HG, FUG, HG were less intense (P<0.05) compared to the findings in the rats in CCG. In conclusion, mad honey and 5-FU reduced anaplastic cell growth and oxidative stress via suppressed anti-apoptotic activity. Considering the histopathological findings in the liver and kidney, no toxicity occurred related to mad honey and 5-FU metabolization. Therefore, the combined use of these two substances may be an alternative method in the treatment of colon cancer.

Keywords

• Apoptosis
• Colon Cancer
• Oxidative Stress
• Turkish Mad Honey

References

1. Afrin S, Giampieri F, Cianciosi D, et al (2021): Strawberry tree honey in combination with 5-fluorouracil enhances chemosensitivity in human colon adenocarcinoma cells. Food Chem Toxicol, 156, 112484.
2. Ahmed HH, El-Abhar HS, Hassanin EAK, et al (2017): Punica granatum suppresses colon cancer through downregulation of Wnt/β-Catenin in rat model. Rev Bras Farmacogn, 27, 627-635.
3. Akcilar R, Akcılar A, Koçak C, et al (2015): Effects of Ukrain on intestinal apoptosis caused by ischemia-reperfusion injury in rats. Int J Clin Exp Med, 12, 22158-22166.
4. Al-Hassan OS, Atawodi SE (2019): Chemopreventive effect of dietary inclusion with Crassocephalum rubens (Juss ex Jacq) leaf on N-methyl-N-nitrosourea(MNU)-induced colorectal carcinogenesis in Wistar rats. J Funct Foods, 63, 103589.
5. Alzahrani HA, Boukraa L, Bellik Y, et al (2012): Evaluation of the Antioxidant Activity of Three Varieties of Honey from Different Botanical and Geographical Origins. Glob J Health Sci, 4, 192-196.
6. Aviram M, Rosenblat M (2004): Paraoxonases 1, 2 and 3, oxidative stress, and macrophage foam cell formation during atherosclerosis development. Free Radic Biol Med, 37, 1304–1316.
7. Bilir EK, Tutun H, Sevin S, et al (2018): Cytotoxic effects of Rhododendron ponticum L. extract on prostate carcinoma and adenocarcinoma cell line (DU145, PC3). Kafkas Univ Vet Fak Derg, 24, 451-457.
8. Bilir K, Sevin S, Tutun H, et al (2018): Cytotoxıc And Antı-Prolıferatıve Effects Of Rhododendron Pontıcum L. Extract On Rat Glıoma Cell Lıne (F98). IJPSR, 5, 1815-1820.

9. Biluca FC, da Silva B, Caon T, et al (2020): Investigation of phenolic compounds, antioxidant and anti-inflammatory activities in stingless bee honey (Meliponinae). Food Res Int, 12908756.
10. Cersosimo RJ (2013): Management of advanced colorectal cancer, part 1. Am J Health Syst Pharm, 70, 395-406.
11. De-Souza ASC, Costa-Casagrande TA (2018): Animal models for colorectal cancer. Arq Bras Cir Dig, 31, 1369.
12. Detre S, Saclani JG, Dowsett, M (1995): A "quickscore" method for immunohistochemical semiquantitation: validation for oestrogen receptor in breast carcinomas. Journal of Clinical Pathology, 48, 876–878.
13. Dogan G, İpek H, Bas Y, et al (2019): Experimental study on prophylactic effects of vardenafil in ischemia–reperfusion model with intestinal volvulus injury in rats. J Pediatr Surg, 54, 2172-2177.
14. Dönmez M, Kaya E (2020): Separating Mad Honey from Other Honeys with Grayanotoxin Analysis in LC-MS/MS. International Journal of Traditional and Complementary Medicine Research, 2, 48-53.
15. Dzugan M, Tomczyk M, Sowa P, et al (2018): Antioxidant Activity as Biomarker of Honey Variety. Molecules, 28, 2069.
16. Erejuwa OO, Sulaiman SA, Wahab MSA (2014): Effects of honey and its mechanisms of action on the development and progression of cancer. Molecules, 19, 2497-2522.
17. Faustino-Rocha AI, Ferreira R, Oliveire PA, et al (2020): N-Methyl-N-nitrosourea as a mammary carcinogenic agent. Tumor Biol, 36, 9095-9117.
18. Gal FA, Stan L, Tabaran F, et al (2020): Chemopreventive Effects of Propolis in the MNU-Induced Rat Mammary Tumor Model. Oxid Med Cell Longev, 10, 1-13.
19. Hamza A, Khalil W, Ahmad H (2013): Possible therapeutic mechanisms of turmeric against colo-rectal cancer induced by N-methylnitrosourea in experimental models. J Med Plant Res, 7, 1940-1950.
20. Harrison S, Benziger H (2011): The molecular biology of colorectal carcinoma and its implications: a review. Surgeon, 9, 200-210.
21. Hazilawati H, Hutheyfa A, Rosly S, et al (2010): Haematological Parameters of Leukaemic Rats Supplemented with Morinda Citrifolia. Med J Malaysia, 65, 125-126.
22. Huang Z, Liu CA, Cai PZ, et al (2020): Omega-3PUFA attenuates MNU-induced colorectal cancer in rats by blocking PI3K/AKT/BCL-2 signaling. Onco Targets Ther, 13, 1953-1965.
23. Iliemene UD, Atawodi SE (2019): Preventive potential of dietary inclusion of Brachystegia eurycoma (Harms) seeds on N-methyl-N-nitrosourea-induced colon carcinogenesis in Wistar rats. J Ethnopharmacol, 238, 111858.
24. Issa JP (2008): Colon cancer: it’s CIN or CIMP. Clin Cancer Res, 14, 5939-5940.
25. Issaad FZ, Fernandes IPG, Enache TA, et al (2019): Honey and Pollen Phenolic Composition, Antioxidant Capacity, and DNA Protecting Properties. Electroanalysis, 31, 611.
26. Jisha N, Vysakh A, Vijeesh V, et al (2020): Methanolic Extract of Muntingia Calabura L. Mitigates 1,2-Dimethyl Hydrazine Induced Colon Carcinogenesis in Wistar Rats. Nutr Cancer, 25, 1-13.
27. Larrosa M, Yanéz-Gascón MJ, Selma MV, et al (2009): Effect of a low dose of dietary resveratrol on colon microbiota, inflammation and tissue damage in a DSS-induced colitis rat model. J Agric Food Chem, 57, 2211-2220.
28. Mabrouk GM, Moselhy SS, Zohny SF, et al (2002): Inhibition of methylnitrosourea (MNU) induced oxidative stress and carcinogenesis by orally administered bee honey and Nigella grains in Sprague Dawely rats. J Exp Clin Cancer Res, 21, 341-346.
29. Mahomoodally MF, Sieniawska E, Sinan KI, et al (2020): Utilisation of Rhododendron luteum Sweet bioactive compounds as valuable source of enzymes inhibitors, antioxidant, and anticancer agents. Food Chem Toxicol, 135, 111052.
30. Margaoan R, Topal E, Balkanska R, et al (2021): Monofloral Honeys as a Potential Source of Natural Antioxidants, Minerals and Medicine. Antioxidants, 10, 1023.
31. Osowole AA, Oni A, Onadje FO, et al (2013): Effects of Injected N-methyl-N-nitrosourea (MNU) in Albino Mice on the Histology and Haematology of Selected Organs of the Circulatory, Lymphoid and Digestive Systems. Curr J Appl Sci Techno, 3, 17-33.
32. Oztasan N, Songur A (2007): The Use Of "Mad Honey" As An Antihypertensive Agent In Rats -A Preliminary Study. Med J Kocatepe, 8, 55-58.
33. Rehman MU, Rashid S, Arafah A, et al (2020): Piperine Regulates Nrf-2/Keap-1 Signalling and Exhibits Anticancer Effect in Experimental Colon Carcinogenesis in Wistar Rats. Biology, 9, 302.
34. Sahin TK, Bilir B, Kucuk O (2021): Modulation of inflammation by phytochemicals to enhance efficacy and reduce toxicity of cancer chemotherapy. Crit Rev Food Sci Nutr, 16, 1-15.
35. Udoh IE, Aprioku JS, Siminialayi I (2017): In-vivo Anti-tumor Evaluation of Dihydroartemisinin-Derived Endodisulphide on MNU-induced Liver Cancer in Sprague-Dawley Rats. Int J Trop Dis Health, 25, 1-13.
36. Umesalma S, Sudhandiran G (2010): Differential inhibitory effects of the polyphenol ellagic acid on inflammatory mediators NF-kappaB, iNOS, COX-2, TNF-alpha, and IL-6 in 1,2-dimethylhydrazine-induced rat colon carcinogenesis. Basic Clin Pharmacol Toxicol, 107, 650-655.
37. Usta A, Dede S, Çetin S (2018): Deneysel diyabetli ratlarda timokinon uygulanmasının doku total oksidan ve antioksidan durumuna etkisi. Atatürk Üniversitesi Vet Bil Derg, 13, 84-91.
38. Vdoviakova K, Petrovova E, Maloveska M, et al (2016): Surgical Anatomy of the Gastrointestinal Tract and Its Vasculature in the Laboratory Rat. Gastroenterol Res Pract, 2632368.
39. Yeşil T, Akgül Y (2022): Major components of Rhododendron luteum leaves. Nat Prod Res, 23, 1-5.