Aqueous parsley (Petroselinum crispum) extract ameliorated methotrexate-induced brain and small intestine damage in rats

Abstract

Methotrexate (MTX) is a widely used antiarthritic and chemotherapeutic agent known to cause damage to various tissues. This study investigated the potential protective effects of parsley extract against MTX-induced brain and intestinal tissue damage. Sprague-Dawley rats were divided into control, control + parsley, MTX, and MTX + parsley. MTX (20 mg/kg, i.p.) was administered to the MTX and MTX + parsley groups. The control + parsley, and MTX + parsley groups were administered 2 g/kg parsley extract by oral gavage for five consecutive days. After the fifth day, brain and small intestinal tissues were taken. Total protein, nitric oxide, lipid peroxidation, glutathione levels, tissue factor, superoxide dismutase, and glutathione S-transferase activities were determined in these tissues. The protein profiles of the tissues were evaluated using SDS polyacrylamide gel electrophoresis. Parsley administration caused a decrease in lipid peroxidation levels in both tissues of the MTX group. On the other hand, glutathione level, glutathione-S-transferase, and superoxide dismutase activities were found to be increased. On the other hand, parsley decreased the nitric oxide level which was increased in the intestinal tissues of the MTX group. There was no significant change in brain nitric oxide level and tissue factor activity between groups. MTX and parsley administration altered protein expression, leading to the appearance or disappearance of specific bands in intestinal and brain tissues. In conclusion, parsley alleviated MTX-induced damage in brain and intestinal tissues by reducing lipid peroxidation and modulating antioxidant defenses.

Keywords

• Brain
• Methotrexate
• Oxidative stress
• Parsley
• Small Intestine

References

1. 1. Al-Howiriny T, Al-Sohaibani M, El-Tahir K, et al (2003): Prevention of experimentally-induced gastric ulcers in rats by an ethanolic extract of "Parsley" Petroselinum crispum. Am J Chin Med, 31, 699-711.
2. 2. AlJohani NI (2021): Role of folinic acid in methotrexate-based prophylaxis of graft-versus-host disease following hematopoietic stem cell transplantation. Hematology, 26, 620-27.
3. 3. Ayalon I, Friedman S, Binenbaum Y, et al (2019): A Case of Methotrexate Neurotoxicity Presented as Status Epilepticus, Encephalopathy, and High Fever. J Investig Med High Impact Case Rep, 7, 2324709619862311.
4. 4. Azadnasab R, Kalantar H, Khorsandi L, et al (2021): Epicatechin ameliorative effects on methotrexate-induced hepatotoxicity in mice. Human Exp Toxicol, 40, 603-610.
5. 5. Bajic JE, Johnston IN, Howarth GS, et al (2018): From the Bottom-Up: Chemotherapy and Gut-Brain Axis Dysregulation. Front Behav Neurosci, 12.
6. 6. Beutler E (1984): Glutathione in red blood cell metabolism. 112-114. In: A manuel of biochemical methods. Grune & Stratton, Newyork.
7. 7. Boukhettala N, Leblond J, Claeyssens S, et al (2009): Methotrexate induces intestinal mucositis and alters gut protein metabolism independently of reduced food intake. Am J Physiol Endocrinol Metab, 296, E182-E90.
8. 8. Chu AJ (2011): Tissue factor, blood coagulation, and beyond: an overview. Int J Inflamm, 2011, 367284.

9. 9. DelGiudice LA, White GA (2009): The role of tissue factor and tissue factor pathway inhibitor in health and disease states. J Vet Emerg Crit Care, 19, 23-29.
10. 10. El-Boghdady NA (2011): Protective effect of ellagic acid and pumpkin seed oil against methotrexate-induced small intestine damage in rats. Indian J Biochem Biophys, 48, 380-87.
11. 11. Ertaş B, Turan FB, Özbeyli D, et al (2021): Protective effects of Petroselinum crispum (Parsley) extract against methotrexate-induced hepatotoxicity. Eur J Biol, 80, 173-78.
12. 12. Ertik O, Sacan O, Yanardag R (2023): Anti-adenosine deaminase, anti-neuraminidase, anti-xanthine oxidase, anti-acetylcholinesterase and antioxidant activities of parsley extract. J Herb Med, 42, 100787.
13. 13. Faul F, Erdfelder E, Lang AG, et al (2007): G* Power 3: A flexible statistical power analysis program for the social, behavioral, and biomedical sciences. Behav Res Methods, 39, 175-91.
14. 14. Grover SP, Mackman N (2020): Tissue factor in atherosclerosis and atherothrombosis. Atherosclerosis, 307, 80-86.
15. 15. Gürel A, Kaya K (2022): Bromelain has Antioxidant Effect on Methotrexate Hepatotoxicity and Nephrotoxicity. Van Sağ Bil Derg, 15, 37-42.
16. 16. Habig WH, Jakoby WB (1981): Assays for differentiation of glutathione S-Transferases. Methods Enzymol, 77, 398-405.
17. 17. He F (2011): Laemmli-sds-page. Bio-protocol, e80-e80.
18. 18. Hess JA, Khasawneh MK (2015): Cancer metabolism and oxidative stress: Insights into carcinogenesis and chemotherapy via the non-dihydrofolate reductase effects of methotrexate. BBA Clin, 3, 152-61.
19. 19. Huang X, Fang Q, Rao T, et al (2020): Leucovorin ameliorated methotrexate induced intestinal toxicity via modulation of the gut microbiota. Toxicol Appl Pharmacol, 391, 114900.
20. 20. Ingram G (1976): Reference method for the one stage prothrombin time test on human blood. Thromb Haemost, 36, 237-38.
21. 21. Katturajan R, Vijayalakshmi S, Rasool M, et al (2021): Molecular toxicity of methotrexate in rheumatoid arthritis treatment: A novel perspective and therapeutic implications. Toxicology, 461, 152909.
22. 22. Kraus M, Çetin M, Aricioglu F (2016): The microbiota and gut-brain axis. Psychiatry Behav Sci, 172.
23. 23. Ledwożyw A, Michalak J, Stȩpień A, et al (1986): The relationship between plasma triglycerides, cholesterol, total lipids and lipid peroxidation products during human atherosclerosis. Clin Chim Acta, 155, 275-83.
24. 24. Mahmood S, Hussain S, Malik F (2014): Critique of medicinal conspicuousness of Parsley (Petroselinum crispum): a culinary herb of Mediterranean region. Pak J Pharm Sci, 27, 193-202.
25. 25. Malayeri A, Badparva R, Mombeini MA, et al (2022): Naringenin: a potential natural remedy against methotrexate-induced hepatotoxicity in rats. Drug Chem Toxicol, 45, 491-98.
26. 26. Maodaa SN, Allam AA, Ajarem J, et al (2016): Effect of parsley (Petroselinum crispum, Apiaceae) juice against cadmium neurotoxicity in albino mice (Mus musculus). Behav Brain Funct, 12, 1-16.
27. 27. Miranda KM, Espey MG, Wink DA (2001): A rapid, simple spectrophotometric method for simultaneous detection of nitrate and nitrite. Nitric oxide, 5, 62-71.
28. 28. Mylroie AA, Collins H, Umbles C, et al (1986): Erythrocyte superoxide dismutase activity and other parameters of copper status in rats ingesting lead acetate. Toxicol Appl Pharm, 82, 512-20.
29. 29. Ozcicek F, Kara AV, Akbas EM, et al (2020): Effects of anakinra on the small intestine mucositis induced by methotrexate in rats. Exp Anim, 69, 144-152.
30. 30. Pellacani C, Eleftheriou G (2020): Neurotoxicity of antineoplastic drugs: Mechanisms, susceptibility, and neuroprotective strategies. Adv Med Sci, 65, 265-85.
31. 31. Punoševac M, Radović J, Leković A, et al (2021): A review of botanical characteristics, chemical composition, pharmacological activity and use of parsley. Arch Pharm, 71, 177-96.
32. 32. Rtibi K, Selmi S, Grami D, et al (2018): Methotrexate produces gastrointestinal stress via oxidative stress-caused acute physiological disruptions in water and electrolytes transport in the mucosal intestine. Recent Adv Biol Med, 4, 3762.
33. 33. Schneider CA, Rasband WS, Eliceiri KW (2012): NIH Image to ImageJ: 25 years of image analysis. Nat Methods, 9, 671-75.
34. 34. Shiga S, Machida T, Yanada T, et al (2020): The role of nitric oxide in small intestine differs between a single and a consecutive administration of methotrexate to rats. J Pharmacol Sci, 143, 30-38.
35. 35. Sklyarova YO, Fomenko I (2018): Action of hydrogen sulfide donors on nitroso-oxidative processes in small intestine of rats with methotrexate-induced enteropathy. Med Clin Chem, 50-56.
36. 36. Sritawan N, Suwannakot K, Naewla S, et al (2021): Effect of metformin treatment on memory and hippocampal neurogenesis decline correlated with oxidative stress induced by methotrexate in rats. Biomed Pharmacother, 144, 112280.
37. 37. Subramaniam CB, Bowen JM, Gladman MA, et al (2020): The microbiota-gut-brain axis: An emerging therapeutic target in chemotherapy-induced cognitive impairment. Neurosci Biobehav Rev, 116, 470-79.
38. 38. Suwannakot K, Sritawan N, Naewla S, et al (2022): Melatonin attenuates methotrexate-induced reduction of antioxidant activity related to decreases of neurogenesis in adult rat hippocampus and prefrontal cortex. Oxid Med Cell Longev, 2022, 1596362.
39. 39. Şener G, Ekşioğlu Demiralp E, Çetiner M, et al (2006): L-Carnitine ameliorates methotrexate-induced oxidative organ injury and inhibits leukocyte death. Cell Biol Toxicol, 22, 47-60.
40. 40. Taha M, Eldemerdash OM, Elshaffei IM, et al (2023): Apigenin attenuates hippocampal microglial activation and restores cognitive function in methotrexate-treated rats: Targeting the miR-15a/ROCK-1/ERK1/2 pathway. Mol Neurobiol, 60, 3770-87.
41. 41. Tufan E, Sivas GG, Gürel Gökmen B, et al (2022): Inhibitory effect of whey protein concentrate on SARS-CoV-2-targeted furin activity and spike protein-ACE2 binding in methotrexate-induced lung damage. J Food Biochem, 46, e14039.
42. 42. Vardi N, Parlakpinar H, Ozturk F, et al (2008): Potent protective effect of apricot and β-carotene on methotrexate-induced intestinal oxidative damage in rats. Food Chem Toxicol, 46, 3015-22.
43. 43. Vora SR, Patil RB, Pillai MM (2009): Protective effects of Petroselinum crispum (Mill) Nyman ex AW Hill leaf extract on D-galactose-induced oxidative stress in mouse brain. Ind J Exp Biol, 47, 338-42.
44. 44. Wang XJ, Luo Q, Li T, et al (2022): Origin, evolution, breeding, and omics of Apiaceae: a family of vegetables and medicinal plants. Hortic Res, 9, uhac076.
45. 45. Waterborg JH (2009): The Lowry Method for Protein Quantitation. 7-10. In: JM Walker (Ed). The Protein Protocols Handbook. Humana Press, Totowa, NJ.
46. 46. Welbat JU, Naewla S, Pannangrong W, et al (2020): Neuroprotective effects of hesperidin against methotrexate-induced changes in neurogenesis and oxidative stress in the adult rat. BiochemPharmacol, 178, 114083.
47. 47. Yang H, Liu Y, Cai R, et al (2021): A narrative review of relationship between gut microbiota and neuropsychiatric disorders: mechanisms and clinical application of probiotics and prebiotics. Ann Palliat Med, 10, 2304-13.
48. 48. Yılmaz S, Tufan E, Sivas GG, et al (2022): The effect of whey proteins on the brain and small intestine nitric oxide levels: protein profiles in methotrexate-induced oxidative stress. Experimed, 12, 113-18.