Comparative study of gavage and intraperitoneal administration of gamma-oryzanol in alleviation/attenuation in a rat animal model of renal ischemia/reperfusion-induced injury

Document Type : Original Article


1 Young Researchers and Elite Club, Tabriz Branch, Islamic Azad University, Tabriz, Iran

2 Faculty of Veterinary Medicine, Tabriz Branch, Islamic Azad University, Tabriz, Iran

3 Department of Clinical Sciences, Faculty of Veterinary Medicine, University of Tabriz, Tabriz, Iran

4 Molecular Medicine Research Center, Tabriz University of Medical Sciences, Tabriz, Iran


Objective(s): Ischemia/reperfusion (I/R) is the leading cause of acute kidney injury. This study aimed to elucidate the reno-protective effect of gamma-oryzanol (GO) by comparing gavage and intraperitoneal (IP) administration methods on renal I/R injury in a rat model.   
Materials and Methods: Rats were divided into four groups including (group 1) sham, (group 2) I/R-control, (group 3) I/R+GO gavage-treated, and (group 4) I/R+ GO IP-treated. A single dose of GO was administrated to groups 3 and 4 (100 mg/kg body weight), 60 min before induction of I/R. After anesthesia, I/R was created by 45 min of ischemia, followed by 6 hr of reperfusion.  Then, blood and tissue samples were subjected to evaluation of renal function, anti-oxidant capacity, inflammation, apoptotic proteins, and IKB/NF-kB pathway.
Results: The two GO administration methods showed improvement of renal function along with attenuation of histological abnormalities. An increase in antioxidant capacity along with a decrease in pro-inflammatory markers, decline in the expression levels of BAX, Bax/Bcl-2, and caspase-3, and up-regulation of Bcl-2 expression were recorded. Moreover, a significant decrease in NF-Kb, p-IKBα, and MMP-2/9 with an increase in IKBα levels were also observed.  Overall, in a comparative evaluation between the two gavage and IP administration methods, we did not find any differences in all examined parameters, except IL-6 which had a better result via gavage.
Conclusion: A single dose of GO administration has a reno-protective effect against renal I/R injury. Gavage and IP administration exhibit similar efficiency in alleviation of I/R injury.


1. Kellum JA, Unruh ML, Murugan R. Acute kidney injury. BMJ Clin Evid 2011; 2011:2001-2037.
2. Han SJ, Lee HT. Mechanisms and therapeutic targets of ischemic acute kidney injury. Kidney Res Clin Pract 2019; 38:427-440.
3. Kalogeris T, Baines CP, Krenz M, Korthuis RJ. Cell biology of ischemia/reperfusion injury.  Int Rev Cell Mol 2012;298:229-317.
4. Vishwakarma VK, Upadhyay PK, Gupta JK, Yadav HN. Pathophysiologic role of ischemia reperfusion injury: A review. J Ind Coll 2017;7:97-104.
5. Wu M-y, Yiang G-t, Liao W-T, Tsai AP-Y, Cheng Y-L, Cheng P-W, et al. Current mechanistic concepts in ischemia and reperfusion injury. Cell Physiol Biochem 2018; 46:1650-1667.
6. Martin JL, Gruszczyk AV, Beach TE, Murphy MP, Saeb-Parsy K. Mitochondrial mechanisms and therapeutics in ischaemia reperfusion injury. Pediatr Nephrol 2019; 34:1167-1174.
7. Malek M, Nematbakhsh M. Renal ischemia/reperfusion injury; from pathophysiology to treatment. J Renal Inj Prev 2015;4:20-27.
8. Rovcanin B, Medic B, Kocic G, Cebovic T, Ristic M, Prostran M. Molecular dissection of renal ischemia-reperfusion: oxidative stress and cellular events. Curr Med Chem 2016; 23:1965-1980.
9. Havasi A, Borkan SC. Apoptosis and acute kidney injury. Kidney Int 2011; 80:29-40.
10. Rashtchizadeh N, Argani H, GHORBANI HA, Nezami N, Safa J, Montazersaheb S. C-reactive protein level following treatment and withdrawal of lovastatin in diabetic nephropathy. Iran J Kidney Dis 2009;3:93-98.
11. Ahmadvand H, Babaeenezhad E, Nayeri H, Nezhad ZZ. Selenium effects on anti-oxidant and inflammatory indices in renal ischemia-reperfusion injury in rats. J Renal Inj Prev 2018;8:71-77.
12. Lobo V, Patil A, Phatak A, Chandra N. Free radicals, anti-oxidants and functional foods: Impact on human health. Pharmacogn Rev 2010;4:118-126.
13. Granger DN, Kvietys PR. Reperfusion injury and reactive oxygen species: the evolution of a concept. Redox Biol 2015;6:524-551.
14. Fathi E, Charoudeh HN, Sanaat Z, Farahzadi R. Telomere shortening as a hallmark of stem cell senescence. Stem Cell Investig 2019;6:7-12.
15. Brazvan B, Farahzadi R, Mohammadi SM, Saheb SM, Shanehbandi D, Schmied L, et al. Key immune cell cytokines affects the telomere activity of cord blood cells in vitro. Adv Pharm Bull 2016;6:153-161.
16. Dennis JM, Witting PK. Protective role for anti-oxidants in acute kidney disease. Nutrients 2017;9:718-743.
17. Krishnaiah D, Sarbatly R, Nithyanandam R. A review of the anti-oxidant potential of medicinal plant species. Food Bioprod Process 2011; 89:217-233.
18. Kushwaha R. Pharmacognosy of rice bran oil-A review. Int J Green Pharm 2019;12:784-790.
19. Lai O-M, Jacoby JJ, Leong W-F, Lai W-T. Nutritional Studies of Rice Bran Oil. Rice Bran Oil 2019; 19-54.
20. Tuncel NB, Yılmaz N. Gamma-oryzanol content, phenolic acid profiles and anti-oxidant activity of rice milling fractions. Eur Food Res Technol 2011;233:577-585.
21. Toorani MR, Golmakani M-T, Gahruie HH. Anti-oxidant activity and inhibitory mechanism of γ-oryzanol as influenced by the unsaturation degree of lipid systems. Food Sci Technol 2020;133:109930-109939.
22. de Gomes MG, Donato F, Souza LC, Goes AR, Borges Filho C, Del Fabbro L, et al. γ-Oryzanol supplementation modifies the inflammatory and oxidative response in fulminant hepatic failure in mice. Pharma Nutrition 2018;6:191-197.
23. Ashraf S, Ghufran Saeed SM, Saify ZS, Haider S, Sayed SA, Ali R, et al. Potential nutraceutical benefits of basmati rice bran oil as analgesic, anti-inflammatory and anti-arthritis. Pak J Pharm Sci 2019;32:2545-2551.
24. Islam M, Murata T, Fujisawa M, Nagasaka R, Ushio H, Bari A, et al. Anti‐inflammatory effects of phytosteryl ferulates in colitis induced by dextran sulphate sodium in mice. Br J Pharmacol 2008;154:812-824.
25. Francisqueti FV, Ferron AJT, Hasimoto FK, Alves PHR, Garcia JL, Dos Santos KC, et al. Gamma oryzanol treats obesity-induced kidney injuries by modulating the adiponectin receptor 2/PPAR-α axis. Oxid Med Cell Longev 2018;2018:1-9.
26. Hashemi Feyzabadi SE, Davoodi M, Obeidavi Z. Gamma-oryzanol alleviated adverse effects of DENA-induced oxidative stress in rat kidney. GMJ Med 2019;3:173-183.
27. Petchdee S, Laosripaiboon W, Jarussophon N, Kumphune S. Cardio-protective effects of germinated brown rice extract against myocardial ischemia reperfusion injury. High Blood Press Cardiovasc Prev 2020;27:251-258.
28. Ghatak SB, Panchal SJ. Anti-hyperlipidemic activity of oryzanol, isolated from crude rice bran oil, on Triton WR-1339-induced acute hyperlipidemia in rats. Rev Bras Farmacogn 2012;22:642-648.
29. Kaya C, Karabulut R, Turkyilmaz Z, Sonmez K, Kulduk G, Gülbahar Ö, et al. Lycopene has reduced renal damage histopathologically and biochemically in experimental renal ischemia-reperfusion injury. Ren Fail 2015;37:1390-1395.
30. Melnikov VY, Faubel S, Siegmund B, Lucia MS, Ljubanovic D, Edelstein CL. Neutrophil-independent mechanisms of caspase-1–and IL-18–mediated ischemic acute tubular necrosis in mice. J Clin Investig 2002;110:1083-1091.
31. Uchiyama M, Mihara M. Determination of malonaldehyde precursor in tissues by thiobarbituric acid test. Anal Biochem 1978;86:271-278.
32. Marklund S, Marklund G. Involvement of the superoxide anion radical in the autoxidation of pyrogallol and a convenient assay for superoxide dismutase. Eur J Biochem 1974;47:469-474.
33. Claiborne A. Handbook of methods for oxygen radical research. Florida: CRC Press, Boca Raton 1985.
34. Singh D, Chander V, Chopra K. Carvedilol attenuates ischemia–reperfusion‐induced oxidative renal injury in rats. Fundam Clin Pharmacol 2004;18:627-634.
35. Farahzadi R, Fathi E, Vietor I. Mesenchymal stem cells could be considered as a candidate for further studies in cell-based therapy of alzheimer’s disease via targeting the signaling pathways. ACS Chem Neurosci 2020;11:1424-1435.
36. Fathi E, Valipour B, Sanaat Z, Charoudeh HN, Farahzadi R. Interleukin-6,-8, and TGF-β secreted from mesenchymal stem cells show functional role in reduction of telomerase activity of leukemia cell via Wnt5a/β-catenin and P53 pathways. Adv Pharm Bull 2020;10:307-314.
37. Tarhriz V, Eyvazi S, Musavi M, Abasi M, Sharifi K, Ghanbarian H, et al. Transient induction of Cdk9 in the early stage of differentiation is critical for myogenesis. J Cell Biochem 2019;120:18854-18861.
38. Kinra M, Arora D, Mudgal J, Pai K, Rao CM, Nampoothiri M. Effect of caffeic acid on ischemia-reperfusion-induced acute renal failure in rats. Pharmacology 2019;103:315-319.
39. Yaribeygi H, Simental‐Mendía LE, Butler AE, Sahebkar A. Protective effects of plant‐derived natural products on renal complications. J Cell Physiol 2019; 234:12161-12172.
40. Tanyeli A, Eraslan E, Güler MC, Nezahat K, Akaras N. Gossypin protects against renal Ischemia-Reperfusion Injury in rats. Kafkas Univ Vet Fak Derg 2020;26:89-96.
41. Szcześniak K, Ostaszewski P, Ciecierska A, Sadkowski T. Investigation of nutriactive phytochemical–gamma‐oryzanol in experimental animal models. J Anim Physiol Anim Nutr 2016;100:601-617.
42. Teodoro AJ. Bioactive compounds of food: their role in the prevention and treatment of diseases. Oxid Med Cell Longev 2019; 1-4.
43. Fathi E, Farahzadi R. Zinc sulphate mediates the stimulation of cell proliferation of rat adipose tissue-derived mesenchymal stem cells under high intensity of EMF exposure. Biol Trace Elem Res 2018;184:529-535.
44. Erol N, Saglam L, Saglam YS, Erol HS, Altun S, Aktas MS, et al. The protection potential of anti-oxidant vitamins against acute respiratory distress syndrome: a rat trial. Inflammation 2019;42:1585-1594.
45. Yaribeygi H, Farrokhi FR, Rezaee R, Sahebkar A. Oxidative stress induces renal failure: A review of possible molecular pathways. J Cell Biochem 2018;119:2990-2998.
46. Zolali E, Asgharian P, Hamishehkar H, Kouhsoltani M, Khodaii H, Hamishehkar H. Effects of gamma oryzanol on factors of oxidative stress and sepsis-induced lung injury in experimental animal model. Iran J Basic Med Sci 2015;18:1257-1263.
47. Thurman JM. Triggers of inflammation after renal ischemia/reperfusion. Clin Immunol 2007;123:7-13.
48. Falcão TR, de Araújo AA, Soares LAL, de Moraes Ramos RT, Bezerra ICF, Ferreira MRA, et al. Crude extract and fractions from Eugenia uniflora Linn leaves showed anti-inflammatory, anti-oxidant, and antibacterial activities. BMC Complement Altern Med 2018;18:84-96.
49. Liu Y, Shi B, Li Y, Zhang H. Protective effect of luteolin against renal ischemia/reperfusion injury via modulation of pro-inflammatory cytokines, oxidative stress and apoptosis for possible benefit in kidney transplant. Med Sci Mon Int Med J Exp Clin Res 2017;23:5720-5727.
50. Van Opdenbosch N, Lamkanfi M. Caspases in cell death, inflammation, and disease. Immunity 2019; 50:1352-1364.
51. Loppnow H, Guzik K, Pryjma J. The role of caspases in modulation of cytokines and other molecules in apoptosis and inflammation.  Madame Curie Biosc Database. 2013; 2013:1–21.
52. Daemen MA, de Vries B, Buurman WA. Apoptosis and inflammation in renal reperfusion injury. Transplantation 2002; 73:1693-1700.
53. Bonegio R, Lieberthal W. Role of apoptosis in the pathogenesis of acute renal failure. Curr Opin Nephrol Hypertens 2002;11:301-308.
54. Borkan SC, editor The role of BCL-2 family members in acute kidney injury. Semin Nephrol 2016;36:237-250.
55. Sanz AB, Sanchez-Niño MD, Ramos AM, Moreno JA, Santamaria B, Ruiz-Ortega M, et al. NF-κB in renal inflammation. J Am Soc Nephrol 2010;21:1254-1262.
56. Gao D, Jing S, Zhang Q, Wu G. Pterostilbene protects against acute renal ischemia-reperfusion injury and inhibits oxidative stress, inducible nitric oxide synthase expression, and inflammation in rats via the Toll‑like receptor 4/nuclear factor‑κB signaling pathway. Exp Ther Med 2018;15:1029-1035.
57. Zhang H, Deng A, Zhang Z, Yu Z, Liu Y, Peng S, et al. The protective effect of epicatechin on experimental ulcerative colitis in mice is mediated by increasing antioxidation and by the inhibition of NF-κB pathway. Pharmacol Rep 2016;68:514-520.
58. Pevzner I, Zorova L, Galkin F, Plotnikov EY, Zorov D. Mitochondria-associated matrix metalloproteinases 2 and 9 in acute renal pathologies. Bull Exp Biol Med 2019;166:334-338.