The Prophylactic Role of Chrysin against Clonazepam Induced Brain Toxicity in Male Albino Rats

Rehab M. Mosaad

Department of Biology, College of Science, Majmaah University, Majmaah 11952, Saudi Arabia and Department of Zoology, Women's College, Ain Shams University, Egypt.

Marwan A. Ibrahim *

Department of Biology, College of Science, Majmaah University, Majmaah 11952, Saudi Arabia.

Hend A. Sabry

Department of Zoology, Women's College, Ain Shams University, Egypt.

*Author to whom correspondence should be addressed.


Background: The bioactive substance chyrus is found in bee propolis and all plants of the species Passiflora. It is well recognized to have neuroprotective properties and a wide range of pharmacological activity.

Purpose: In this work, the consequence of Chrysin administration (50 mg/kg b.wt/day) on brain toxicity caused by Clonazepam (CZP, 2mg/kg b.wt/day) were investigated by measuring NaK ATPase, neuronal oxidative stress, neuro- inflammation, and DNA fragmentation.

Study Design: In our investigation, we used male albino rats 4 weeks old and weighing 60 ± 5 g. There were four groups of ten rats apiece: Group 1: the control group which treated a vehicle with 1% w/v Tween 80. Group 2: given 1% w/v Tween 80-suspended Clonazepam (CZP) at a dose of 2 mg/kg b.wt./day. In Group 3: Chrysin suspended in 1% w/v Tween 80 was given at a rate of 50 mg/kg b.wt/day. Group 4: Clonazepam (CZP) and Chrysin were given at the same prior dosages as before (2 mg/kg b.wt/day for Clonazepam (CZP) and 50 mg/kg b.wt/day for Chrysin).

Methods: Malondialdehyde, nitric oxide, DNA fragmentation, sodium oxide dismutase, catalase, and Na-K ATPase contents were estimated.

Results: According to the biochemical analysis, after the Clonazepam therapy, the brain's contents of malondialdehyde (MDA), nitric oxide (NO), and DNA fragmentation increased, while those of superoxide dismutase (SOD), catalase (CAT), and NaK ATPase activities declined.

Conversely, the biochemical screening of animal brain tissue administered with CZP+ Chyrsin revealed an improvement in the brain tissue's ability to withstand the damage caused by CZP.

Keywords: Chrysin, clonazepam, male albino rats, neurotoxicity

How to Cite

Mosaad, R. M., Ibrahim, M. A., & Sabry, H. A. (2024). The Prophylactic Role of Chrysin against Clonazepam Induced Brain Toxicity in Male Albino Rats. UTTAR PRADESH JOURNAL OF ZOOLOGY, 45(12), 261–268.


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Cunha-Oliveira T, et al. Neurotoxicity of heroin–cocaine combinations in rat cortical neurons. Toxicology. 2010;276(1):11-17.

Ahmed GK, et al. Effect of long-term administration of clonazepam, carbamazepine, and valproate on cognitive, psychological, and personality changes in adult epilepsy: A case–control study. Middle East Current Psychiatry. 2021;28:1-10.

E Nardi A, et al. Clonazepam for the treatment of panic disorder. Current Drug Targets. 2013;14(3):353-364.

Negrusz A, et al. Elimination of 7-aminoclonazepam in urine after a single dose of clonazepam. Analytical and Bioanalytical Chemistry. 2003;376:1198-1204.

Roberts SM, Bezinover DS, Janicki PK. Reappraisal of the role of dolasetron in prevention and treatment of nausea and vomiting associated with surgery or chemotherapy. Cancer Management and Research. 2012;67-73.

Brandt J, et al. Prescribing and deprescribing guidance for benzodiazepine and benzodiazepine receptor agonist use in adults with depression, anxiety, and insomnia: An international scoping review. Eclinicalmedicine. 2024;70.

Longo LP, Johnson B. Addiction: Part I. Benzodiazepines—side effects, abuse risk and alternatives. American Family Physician. 2000;61(7):2121-2128.

Campbell TJ, et al. The epidemiology of benzodiazepine-related toxicity in Ontario, Canada: A population-based descriptive study. Canadian Journal of Public Health. 2023;114(6):956-966.

Gil Tejedor AM, et al. Selective Extraction of Diazepam and Its Metabolites from Urine Samples by a Molecularly Imprinted Solid-Phase Extraction (MISPE) Method. Polymers. 2024;16(5):635.

Mosaad RM, Samir A, Ibrahim HM. Median lethal dose (LD50) and cytotoxicity of Adriamycin in female albino mice. Journal of Applied Pharmaceutical Science. 2017;7(3):77-80.

Ibrahim HM, et al. Preparation of chitosan antioxidant nanoparticles as drug delivery system for enhancing of anti-cancer drug. In Key Engineering Materials; 2018.

Hong JS, et al. Antinociceptive effect of chrysin in diabetic neuropathy and formalin-induced pain models. Animal Cells and Systems. 2020;24(3): 143-150.

Mahgoub AA, et al. Nematocidal activity of chitosan nanoparticles conjugated with albendazole against the enteral and parenteral phases of trichinosis in experimentally infected mice. Journal of Parasitic Diseases; 2024.

Ibrahim MA, et al. A Review of Chitosan and Chitosan Nanofiber: Preparation, Characterization, and Its Potential Applications. Polymers. 2023;15(13).

Mosaad RM, et al. Enhancement of Antimicrobial and Dyeing Properties of Cellulosic Fabrics via Chitosan Nanoparticles. Polymers. 2022;14(19).

Farkhondeh T, et al. Effect of chrysin on nociception in formalin test and serum levels of noradrenalin and corticosterone in rats. International Journal of Clinical and Experimental Medicine. 2015;8(2):2465.

Samarghandian S, et al. Assessment the effect of saffron ethanolic extract (Crocus sativus L.) on oxidative damages in aged male rat liver. Der Pharm Lett. 2016;8(3):283-90.

Samarghandian S, et al. Chrysin treatment improves diabetes and its complications in liver, brain, and pancreas in streptozotocin-induced diabetic rats. Canadian Journal of Physiology and Pharmacology. 2016;94(4):388-393.

Samarghandian S, et al. Protective effects of carvacrol against oxidative stress induced by chronic stress in rat’s brain, liver, and kidney. Biochemistry Research International; 2016.

Williams CA, et al. Chrysin and other leaf exudate flavonoids in the genus Pelargonium. Phytochemistry. 1997;46(8): 1349-1353.

Stompor-Gorący M, Bajek-Bil A, Machaczka M. Chrysin: Perspectives on contemporary status and future possibilities as pro-health agent. Nutrients. 2021;13(6):2038.

Alavi Dana SMM, et al. Chrysin effect against gastric cancer: Focus on its molecular mechanisms. Current Molecular Pharmacology. 2023;16(7):707-711.

Anandhi R, et al. Antihypercholesterolemic and antioxidative effects of an extract of the oyster mushroom, Pleurotus ostreatus, and its major constituent, chrysin, in Triton WR-1339-induced hypercholesterolemic rats. Journal of Physiology and Biochemistry. 2013;69:313-323.

Tong L, et al. Simultaneous determination of baicalin, wogonoside, baicalein, wogonin, oroxylin A and chrysin of Radix scutellariae extract in rat plasma by liquid chromatography tandem mass spectrometry. Journal of Pharmaceutical and Biomedical Analysis. 2012;70:6-12.

Lapidot T, Walker MD, Kanner J. Antioxidant and prooxidant effects of phenolics on pancreatic β-cells In vitro. Journal of Agricultural and Food Chemistry M. M. 2002;50(25):7220-7225.

Rashed AL, Al-Saeed HF, El-Shawwa. Neuroprotective effects of chrysin on adult male albino rats exposed to acrylamide and radiation. Al-Azhar Medical Journal. 2016;45(3):493-504.

He XL, et al. Chrysin improves cognitive deficits and brain damage induced by chronic cerebral hypoperfusion in rats. European Journal of Pharmacology. 2012;680(1-3):41-48.

Mercer LD, et al. Dietary polyphenols protect dopamine neurons from oxidative insults and apoptosis: Investigations in primary rat mesencephalic cultures. Biochemical Pharmacology. 2005;69(2):339-345.

Falbo F, Aiello F. Chrysin: A polyedric flavone as a tool to explore new phytotherapeutic applications and drug design. Archiv der Pharmazie. 2023;356(2):2200347.

El Khashab IH, et al. Chrysin attenuates global cerebral ischemic reperfusion injury via suppression of oxidative stress, inflammation and apoptosis. Biomedicine and Pharmacotherapy. 2019;112:108619.

Mohamed TM, Ghaffar HMA, El Husseiny RMR. Effects of tramadol, clonazepam, and their combination on brain mitochondrial complexes. Toxicology and Industrial Health. 2015;31(12):1325-1333.

Socała K, et al. Effect of tadalafil on seizure threshold and activity of antiepileptic drugs in three acute seizure tests in mice. Neurotoxicity Research. 2018;34:333-346.

Mehri S, et al. Chrysin reduced acrylamide-induced neurotoxicity in both In vitro and In vivo assessments. Iranian Biomedical Journal. 2014;18(2):101.

Reagan‐Shaw S, Nihal M, Ahmad N. Dose translation from animal to human studies revisited. The FASEB Journal. 2008;22(3):659-661.

Ohkawa H. Assay for lipid peroxidation in animal tissues by thiobarbituric acid reaction. Anal Biochem. 1979;44:276-278.

Nims RW, et al. Colorimetric methods for the determination of nitric oxide concentration in neutral aqueous solutions. Methods. 1995;7(1):48-54.

Mikulecká A, et al. Consequences of early postnatal benzodiazepines exposure in rats. II. Social behavior. Frontiers in Behavioral Neuroscience. 2014;8:169.

Kubová H, et al. Neonatal Clonazepam administration induced long-lasting changes in GABAA and GABAB receptors. International Journal of Molecular Sciences. 2020;21(9):3184.

Sabry SA, Ibrahim MA, Mosaad RM. Biochemical, histological and ultrastructural studies on the ameliorative effects of chrysin on the hepatotoxicity of clonazepam in developing male albino rats. African Journal of Biological Sciences. 2022;18(2):161-175.

Lakshmi D, et al. Ameliorating effect of fish oil on acrylamide induced oxidative stress and neuronal apoptosis in cerebral cortex. Neurochemical Research. 2012;37:1859-1867.

Nixon BJ, et al. Chronic exposure to acrylamide induces DNA damage in male germ cells of mice. Toxicological Sciences. 2012;129(1):135-145.

Neophytou CM, et al. In vivo Investigation of the Effect of Dietary Acrylamide and Evaluation of Its Clinical Relevance in Colon Cancer. Toxics. 2023;11(10): 856.

Zhou QG, et al. Neuronal nitric oxide synthase and affective disorders. IBRO Reports. 2018;5:116-132.

Pitsikas N, Nitric Oxide (NO) Synthase Inhibitors: Potential Candidates for the Treatment of Anxiety Disorders? Molecules. 2024;29(6):1411.

Pitsikas N. The role of nitric oxide (NO) donors in anxiety. Lights and shadows. Nitric Oxide. 2018;77:6-11.

Papi S, Ahmadizar F, Hasanvand A. The role of nitric oxide in inflammation and oxidative stress. Immunopathologia Persa. 2019;5(1):e08-e08.

Badawy S, et al. Effects of dependence of tramadol, diazepam and their combination on the brain of Albino rats: Biochemical, histological and immunohistochemical study. Ain Shams Journal of Forensic Medicine and Clinical Toxicology. 2014;23(2):139-147.

Elsukary AE, et al. Comparative study of the Neurotoxic Effects of Pregabalin versus Tramadol in Rats. Neurotoxicity Research. 2022;40(5):1427-1439.

Girgis N, et al. Cellular and DNA changes due to clonazepam abuse in brains of albino rats and role of clonidine during withdrawal period. Mansoura Journal of Forensic Medicine and Clinical Toxicology. 2010;18(1):25-49.

Bittigau P, et al. Antiepileptic drugs and apoptotic neurodegeneration in the developing brain. Proceedings of the National Academy of Sciences. 2002;99(23):15089-15094.

Khan T, et al. Mitochondrial Dysfunction: Pathophysiology and Mitochondria-Targeted Drug Delivery Approaches. Pharmaceutics. 2022;14(12):2657.

Nadanaciva S, et al. Mitochondrial impairment by PPAR agonists and statins identified via immunocaptured OXPHOS complex activities and respiration. Toxicology and Applied Pharmacology. 2007;223(3):277-287.

Patel M. Mitochondrial dysfunction and oxidative stress: Cause and consequence of epileptic seizures. Free Radical Biology and Medicine. 2004;37(12):1951-1962.

Anand KV, et al. Protective role of chrysin against oxidative stress in d‐galactose‐induced aging in an experimental rat model. Geriatrics and Gerontology International. 2012;12(4):741-750.

Yao Y, et al. Chrysin protects against focal cerebral ischemia/reperfusion injury in mice through attenuation of oxidative stress and inflammation. International Journal of Molecular Sciences. 2014; 15(11):20913-20926.

Shang J, et al. Chrysin protects against cerebral ischemia-reperfusion injury in hippocampus via restraining oxidative stress and transition elements. Biomedicine and Pharmacotherapy. 2023;161:114534.

Rashno M, et al. Possible mechanisms involved in the neuroprotective effects of chrysin against mild traumatic brain injury-induced spatial cognitive decline: An In vivo study in a rat model. Brain Research Bulletin. 2023;204: 110779.

Gresa‐Arribas N, et al. Inhibition of CCAAT/enhancer binding protein δ expression by chrysin in microglial cells results in anti‐inflammatory and neuroprotective effects. Journal of Neurochemistry. 2010;115(2):526-536.

Souza LC, et al. Flavonoid Chrysin prevents age-related cognitive decline via attenuation of oxidative stress and modulation of BDNF levels in aged mouse brain. Pharmacology Biochemistry and Behavior. 2015;134:22-30.