Evaluation of the activity of geraniol isolated from lemongrass (Cymbopogon commutatus Stapf.) on ochratoxin A-induced nephrotoxicity: role of the pPI3K/AKT-Nrf2 signaling pathway

Ochratoxin A (OTA) is a mycotoxin that causes immunotoxicity, teratogenicity, hepatotoxicity and nephrotoxicity in humans and animals. Numerous studies have suggested that oxidative stress may increase OTA's nephrotoxicity. Geraniol (GNL), a monoterpene found in many plant oils is an antioxidant and free radical scavenger that helps repair multiple types of tissue damage. OTA-induced nephrotoxicity in mice was assessed using GNL as a protective natural compound. The Swiss albino mice (six to eight weeks old, 25-30g weight) were divided into four groups: control (normal saline), OTA (OTA 5 mg/wt), GNL (GNL 40 mg/wt), and GNL + OTA (OTA 5 mg/wt, 4 h later). Animals were tested for 42 days. Evaluation using body weight, kidney weight, spleen weight, H&E staining for tissue pathology, biochemical markers (Alanine transaminase (ALT), Aspartate transaminase (AST), creatinine, Blood Urea nitrogen (BUN), Western blot, DNA fragmentation), and oxidative markers (malondialdehyde (MDA), superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidase (GPx) has been performed. A significant decrease in body weight was observed after exposure to OTA, while a significant augmentation in spleen weight was noticed. As a result, tissue concentrations of SOD, CAT, and GPx were decreased, while serum concentrations of marker enzymes (ALT, AST, BUN, creatinine and tissue MDA) were increased. In mice, GNL improved enzyme and antioxidant levels. OTA-induced renal injury was prevented by GNL based on H&E tissue pathology. The OTA group also upregulated cleaved caspase-3 and DNA fragmentation, while downregulating pPI3K, pAKT, Nrf2, and Bcl2 protein expression. GNL increased the expression of pPI3K, pAKT, Nrf2, Bcl2, and decreased cleaved caspase-3. Based on these results, GNL protects nephrons via the pPI3K/AKT-Nrf2 signaling pathway. The molecular of OTA-induced renal injury and how GNL protects the kidneys was explained in this study.

Crupi R, Palma E, Siracusa R, et al. Protective effect of Hydroxytyrosol against oxidative stress induced by the ochratoxin in kidney cells: In vitro and in vivo study. Front Vet Sci 2020;7:136. DOI: https://doi.org/10.3389/fvets.2020.00136

Cui G, Li L, Xu W, et al. Astaxanthin protects ochratoxin a-induced oxidative stress and apoptosis in the heart via the Nrf2 pathway. Oxid Med Cell Longev 2020;2020:7639109. DOI: https://doi.org/10.1155/2020/7639109

Rajendran P, Alzahrani AM, Priya Veeraraghavan V, Ahmed EA. Anti-apoptotic effect of flavokawain A on ochratoxin-A-induced endothelial cell injury by attenuation of oxidative stress via PI3K/AKT-mediated Nrf2 signaling cascade. Toxins 2021;13:745. DOI: https://doi.org/10.3390/toxins13110745

Lan M, Zhang Y, Wan X, et al. Melatonin ameliorates ochratoxin A-induced oxidative stress and apoptosis in porcine oocytes. Environ Pollut 2020; 256: 113374. DOI: https://doi.org/10.1016/j.envpol.2019.113374

Elhady MA, Khalaf AAA, Ibrahim MA, et al. Protective effects of fermentation extract against ochratoxin A-induced nephrotoxicity and immunotoxicity in broiler chickens. J Vet Res 2022;66:167-77. DOI: https://doi.org/10.2478/jvetres-2022-0030

Tao Y, Xie S, Xu F, et al. Ochratoxin A: Toxicity, oxidative stress and metabolism. Food Chem. Toxicol 2018;112:320-31. DOI: https://doi.org/10.1016/j.fct.2018.01.002

Rajendran P, Alzahrani AM. Fucoxanthin suppresses OxLDL-induced inflammation via activation of Nrf2 and inhibition of NF-κB signaling. Asian Pac J Trop Biomed 2022;12:207. DOI: https://doi.org/10.4103/2221-1691.343388

Rajendran P, Alzahrani AM, Ahmed EA, Veeraraghavan VP. Kirenol inhibits B[a]P-induced oxidative stress and apoptosis in endothelial cells via modulation of the Nrf2 signaling pathway. Oxid Med Cell Longev 2021;2021:5585303. DOI: https://doi.org/10.1155/2021/5585303

Alzahrani AM, Rajendran P, Veeraraghavan VP, Hanieh H. Cardiac protective effect of Kirenol against doxorubicin-induced cardiac hypertrophy in H9c2 cells through Nrf2 signaling via PI3K/AKT pathways. Int J Mol Sci 2021;22:3269. DOI: https://doi.org/10.3390/ijms22063269

Ismail MB, Rajendran P, AbuZahra HM, Veeraraghavan VP. Mangiferin inhibits apoptosis in doxorubicin-induced vascular endothelial cells via the Nrf2 signaling pathway. Int J Mol Sci 2021;22:4259. DOI: https://doi.org/10.3390/ijms22084259

Kma L, Baruah TJ. The interplay of ROS and the PI3K/Akt pathway in autophagy regulation. Biotechnol Appl Biochem 2022;69:248-264 DOI: https://doi.org/10.1002/bab.2104

Rahmani S, Naraki K, Roohbakhsh A, et al. The protective effects of rutin on the liver, kidneys, and heart by counteracting organ toxicity caused by synthetic and natural compounds. Food Sci Nutr 2023;11:39-56. DOI: https://doi.org/10.1002/fsn3.3041

Ceyhan E, Canbek M. Determining the effects of geraniol on liver regeneration via the nuclear factor κB pathway after partial hepatectomy. Altern Ther Health Med 2017;23: 38-45.

Jayachandran M, Chandrasekaran B, Namasivayam N. Geraniol attenuates oxidative stress by Nrf2 activation in diet-induced experimental atherosclerosis. J Basic Clin Physiol Pharmacol 2015;26:335-46. DOI: https://doi.org/10.1515/jbcpp-2014-0057

Ahmad ST, Arjumand W, Seth A, et al. Preclinical renal cancer chemopreventive efficacy of geraniol by modulation of multiple molecular pathways. Toxicology 2011;290:69-81. DOI: https://doi.org/10.1016/j.tox.2011.08.020

Kandeil MA, Mahmoud MO, Abdel-Razik ARH, Gomaa SB. Thymoquinone and geraniol alleviate cisplatin-induced neurotoxicity in rats through downregulating the p38 MAPK/STAT-1 pathway and oxidative stress. Life Sci 2019;228:145-51. DOI: https://doi.org/10.1016/j.lfs.2019.04.065

Younis NS, Elsewedy HS, Soliman WE, et al. Geraniol isolated from lemon grass to mitigate doxorubicin-induced cardiotoxicity through Nrf2 and NF-κB signaling. Chem Biol Interact 2021;347:109599. DOI: https://doi.org/10.1016/j.cbi.2021.109599

Bradford MM. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 1976;72:248-54. DOI: https://doi.org/10.1006/abio.1976.9999

Misra HP, Fridovich I. The role of superoxide anion in the autoxidation of epinephrine and a simple assay for superoxide dismutase. J Biol Chem 1972;247:3170-75. DOI: https://doi.org/10.1016/S0021-9258(19)45228-9

Rotruck JT, Pope AL, Ganther HE, et al. Selenium: Biochemical role as a component of glutathione peroxidase. Science 1973;179:588-90. DOI: https://doi.org/10.1126/science.179.4073.588

Beutler E. Improved method for the determination of blood glutathione. J Lab Clin Med 1963;61:882-8.

Farombi E, Tahnteng J, Agboola A, et al. Chemoprevention of 2-acetylaminofluorene-induced hepatotoxicity and lipid peroxidation in rats by kolaviron - a Garcinia kola seed extract. Food Chem Toxicol 2000;38:535-41. DOI: https://doi.org/10.1016/S0278-6915(00)00039-9

Alzahrani, AM, Peramiyan R, Bekhet MG, et al. Protective effect of 5, 4'-dihydroxy-6, 8-dimethoxy7-O-rhamnosylflavone from Indigofera aspalathoides Vahl on lipopolysaccharide-induced intestinal injury in mice. Inflammopharmacology 2024;32:3537-51. DOI: https://doi.org/10.1007/s10787-024-01530-y

Lei Y, Fu P, Jun X, Cheng P. Pharmacological properties of geraniol - A review. Planta med 2019;85:48-55.

Lorenzi V, Muselli A, Bernardini AF, et al. Geraniol restores antibiotic activities against multidrug-resistant isolates from gram-negative species. Antimicrob Agents Chemother 2009;53:2209-11. DOI: https://doi.org/10.1128/AAC.00919-08

Ben Ammar R. Potential effects of geraniol on cancer and inflammation-related diseases: A review of the recent research findings. Molecules (Basel, Switzerland) 2023;28:3669. DOI: https://doi.org/10.3390/molecules28093669

Ben Ammar R, Mohamed ME, Alfwuaires M, et al. Anti-inflammatory activity of geraniol isolated from lemon grass on Ox-LDL-stimulated endothelial cells by upregulation of heme oxygenase-1 via PI3K/Akt and Nrf-2 signaling pathways. Nutrients 2022;14:4817. DOI: https://doi.org/10.3390/nu14224817

Li L, Chen Y, Jiao D, et al. Protective effect of astaxanthin on ochratoxin A-induced kidney injury to mice by regulating oxidative stress-related Nrf2/Keap1 pathway. Molecules (Basel, Switzerland) 2020;25:1386. DOI: https://doi.org/10.3390/molecules25061386

Wei W, Liu C, Ke P, et al. Toxicological and physiological effects of successive exposure to Ochratoxin A at food regulatory limits. Food Chem Toxicol 2021;151:112128. DOI: https://doi.org/10.1016/j.fct.2021.112128

Malekinejad H, Farshid A, Mirzakhani N. Liquorice plant extract reduces ochratoxin A-induced nephrotoxicity in rats. Exp Toxicol Pathol 2011;63:125-30. DOI: https://doi.org/10.1016/j.etp.2009.10.006

Pyo MC, Chae SA, Yoo HJ, Lee KW. Ochratoxin A induces epithelial-to-mesenchymal transition and renal fibrosis through TGF-β/Smad2/3 and Wnt1/β-catenin signaling pathways in vitro and in vivo. Arch Toxicol 2020;94:3329-42. DOI: https://doi.org/10.1007/s00204-020-02829-9

Elblehi SS, Hafez MH, El-Sayed YS. L-α-Phosphatidylcholine attenuates mercury-induced hepato-renal damage through suppressing oxidative stress and inflammation. Environ Sci Pollut Res 2019;26:9333-42. DOI: https://doi.org/10.1007/s11356-019-04395-9

Offor SJ, Mbagwu HO, Orisakwe OE. Lead induced hepato-renal damage in male albino rats and effects of activated charcoal. Front Pharmacol 2017;8:107. DOI: https://doi.org/10.3389/fphar.2017.00107

Lei Y, Fu P, Jun X, Cheng P. Pharmacological properties of geraniol – A review. Planta Med 2019;85:48-55. DOI: https://doi.org/10.1055/a-0750-6907

Zuin M, Capatti E, Borghi C, et al. Malondialdehyde levels in hypertensive patients: A non-invasive marker of oxidative stress. A systematic review and meta-analysis. High Blood Press Cardiovasc Prev 2022;29:1-11. DOI: https://doi.org/10.1007/s40292-022-00514-9

Rajendran P, Nandakumar N, Rengarajan T, et al. Antioxidants and human diseases. Clin Chim Acta 2014; 436: 332-47. DOI: https://doi.org/10.1016/j.cca.2014.06.004

Tiwari M, Kakkar P. Plant derived antioxidants–geraniol and camphene protect rat alveolar macrophages against t-BHP induced oxidative stress. Toxicol In Vitro 2009;23:295-301. DOI: https://doi.org/10.1016/j.tiv.2008.12.014

Hosseini SM, Hejazian LB, Amani R, Siahchehreh Badeli N. Geraniol attenuates oxidative stress, bioaccumulation, serological and histopathological changes during aluminum chloride-hepatopancreatic toxicity in male Wistar rats. Environ Sci Pollut Res 2020;27:20076-89. DOI: https://doi.org/10.1007/s11356-020-08128-1

Nezu M, Suzuki N. Roles of Nrf2 in protecting the kidney from oxidative damage. Int J Mol Sci 2020; 21: 2951. DOI: https://doi.org/10.3390/ijms21082951

Ruiz S, Pergola PE, Zager RA, et al. Targeting the transcription factor Nrf2 to ameliorate oxidative stress and inflammation in chronic kidney disease. Kidney Int 2013;83:1029-41. DOI: https://doi.org/10.1038/ki.2012.439

Shelton LM, Park BK, Copple IM. Role of Nrf2 in protection against acute kidney injury. Kidney Int 2013; 84: 1090-95. DOI: https://doi.org/10.1038/ki.2013.248

Dodson M, De La Vega MR, Cholanians AB, et al. Modulating NRF2 in disease: timing is everything. Annu Rev Pharmacol Toxicol 2019;59: 555. DOI: https://doi.org/10.1146/annurev-pharmtox-010818-021856

Egea J, González-Rodríguez Á, Gómez-Guerrero C, Moreno JA. Role of Nrf2 in disease: novel molecular mechanisms and therapeutic approaches. Front Pharmacol 2019;10:1149. DOI: https://doi.org/10.3389/fphar.2019.01149

Ruvolo P, Deng X, May W. Phosphorylation of Bcl2 and regulation of apoptosis. Leukemia 2001;15:515-22. DOI: https://doi.org/10.1038/sj.leu.2402090

Alzahrani S, Bekhet G, Ben Ammar R, et al. The inhibitory effect of geraniol on CCL4-induced hepatorenal toxicity in pregnant mice through the PI3K/AKT signaling pathway. Saudi J Med Med Sci 2024;12:17-26 DOI: https://doi.org/10.4103/sjmms.sjmms_225_23

Thomadaki H, Scorilas A. Bcl2 family of apoptosis-related genes: functions and clinical implications in cancer. Crit Rev Clin Lab Sci 2006;43:1-67. DOI: https://doi.org/10.1080/10408360500295626

Mohamed ME, Elmorsy MA, Younis NS. Renal ischemia/reperfusion mitigation via geraniol: The role of Nrf-2/HO-1/NQO-1 and TLR2,4/MYD88/NFκB pathway. Antioxidants 2022;1: 1568. DOI: https://doi.org/10.3390/antiox11081568