Jundishapur Scientific Medical Journal

Jundishapur Scientific Medical Journal

Protective Effects of Crocin and Aerobic Training on Amphetamine-Induced Alterations in AMPK and IL-1β Expression in the Cerebral Cortex of Female Wistar Rats

Document Type : Original Article

Authors
Hossein Ansari Mahyari 1
Alireza Barari 2
Seyed Javad Ziaolhagh 3
1 PhD student, Department of Sports Physiology, Am.C., Islamic Azad University, Amol, Iran
2 Associated Professor, Department of Sports Physiology, Am.C., Islamic Azad University, Amol, Iran.
3 Assistant Professor, Department of Exercise Physiology, Sha.C., Islamic Azad University, Shahrood, Iran.
10.22118/jsmj.2025.447862.3460
Abstract
Background and Objectives Methamphetamine is a potent central nervous system stimulant, and its chronic use leads to neurotoxicity and brain inflammation. This study investigated the effects of aerobic exercise and crocin (the active compound in saffron) on the expression of inflammatory (IL-1β) and metabolic regulatory (AMPK) genes in the cerebral cortex of female Wistar rats exposed to methamphetamine.
Subjects and Methods In this experimental study, 40 rats were divided into five groups: control, methamphetamine, methamphetamine + exercise, methamphetamine + crocin, and methamphetamine + crocin + exercise. The rats underwent treadmill running for 15–30 minutes at a speed of 20–25 meters per minute, three days a week continued for eight weeks. Crocin (40–80 mg/kg) was administered intraperitoneally. Gene expression was measured using quantitative real-time PCR (qRT-PCR). One-way ANOVA was used for statistical analysis.
Results The results showed that methamphetamine administration significantly increased the expression of IL-1β and AMPK. Both aerobic exercise and crocin independently reduced the methamphetamine-induced elevation in IL-1β and AMPK expression (P < 0.05). The combined intervention of exercise and crocin reduced AMPK expression to the extent that no significant difference was observed between the methamphetamine +crocin+exercise group and the healthy control group (P > 0.05).
Conclusion The findings suggest a protective role of crocin and aerobic exercise in reducing inflammation and improving mitochondrial function in methamphetamine-induced brain damage. These interventions may serve as complementary strategies for preventing neurotoxic effects associated with stimulant drug use.
Keywords
Amphetamine
Neuroinflammation
Mitochondrial biogenesis
Crocin
Aerobic training
Subjects
1.   Shrestha P, Katila N, Lee S, Seo JH, Jeong JH, Yook S. Methamphetamine induced neurotoxic diseases, molecular mechanism, and current treatment strategies. Biomedicine & Pharmacotherapy. 2022 Oct 1;154:113591.[ 10.1016/j.biopha.2022.113591 ][PMID]
2.   Omidvari S, Azimzadeh Z, Rashnoo F, Tahmasebinia F, Keramatinia A, Roozbahany NA, Abbaszadeh HA, Darabi S. Molecular mechanisms and treatment strategies for methamphetamine‑induced neurodegeneration, inflammation and neurotoxicity. Acta neurobiologiae experimentalis. 2023 Dec 28;83(4):414-31.[ 10.55782/ane-2023-2488 ][PMID]
3.   Nemeth DP, Quan N. Modulation of neural networks by interleukin-1. Brain Plasticity. 2021 Jun;7(1):17-32. .[ 10.3233/BPL-200109 ][PMID]
4.   McConnell SE. Methamphetamine-induced Neuroinflammation and Neurotoxicity: a Role for Interleukin-1β (Doctoral dissertation, University of Rochester).
5.   Liśkiewicz A, Przybyła M, Park M, Liśkiewicz D, Nowacka-Chmielewska M, Małecki A, Barski J, Lewin-Kowalik J, Toborek M. Methamphetamine-associated cognitive decline is attenuated by neutralizing IL-1 signaling. Brain, behavior, and immunity. 2019 Aug 1;80:247-54.[ 10.1016/j.bbi.2019.03.016 ][PMID]
6.   Shin EJ, Tran HQ, Nguyen PT, Jeong JH, Nah SY, Jang CG, Nabeshima T, Kim HC. Role of mitochondria in methamphetamine-induced dopaminergic neurotoxicity: involvement in oxidative stress, neuroinflammation, and pro-apoptosis—a review. Neurochemical research. 2018 Jan;43(1):66-78.[ 10.1007/s11064-017-2318-5 ][PMID]
7.   Oh TS, Cho H, Cho JH, Yu SW, Kim EK. Hypothalamic AMPK-induced autophagy increases food intake by regulating NPY and POMC expression. Autophagy. 2016 Nov 1;12(11):2009-25 .[ 10.1080/15548627.2016.1215382 ][PMID]
8.   Kohno D, Sone H, Tanaka S, Kurita H, Gantulga D, Yada T. AMP-activated protein kinase activates neuropeptide Y neurons in the hypothalamic arcuate nucleus to increase food intake in rats. Neuroscience Letters. 2011 Jul 25;499(3):194-8.[ 10.1016/j.neulet.2011.05.060 ][PMID]
9.   Ronnett GV, Ramamurthy S, Kleman AM, Landree LE, Aja S. AMPK in the brain: its roles in energy balance and neuroprotection. Journal of neurochemistry. 2009 May;109:17-23.[ 10.1111/j.1471-4159.2009.05916.x ][PMID]
10. Muraleedharan R, Dasgupta B. AMPK in the brain: its roles in glucose and neural metabolism. The FEBS journal. 2022 Apr;289(8):2247-62 .[ 10.1111/febs.16151 ][PMID]
11. Mozaffaritabar S, Koltai E, Zhou L, Bori Z, Kolonics A, Kujach S, Gu Y, Koike A, Boros A, Radák Z. PGC-1α activation boosts exercise-dependent cellular response in the skeletal muscle. Journal of physiology and biochemistry. 2024 May;80(2):329-35 .[ 10.1007/s13105-024-01006-1 ][PMID]
12. Wu X, Li C, Ke C, Huang C, Pan B, Wan C. The activation of AMPK/PGC-1α/GLUT4 signaling pathway through early exercise improves mitochondrial function and mitigates ischemic brain damage. Neuroreport. 2024 Jul 1;35(10):648-56.[ 10.1097/WNR.0000000000002048 ][PMID]
13. Thirupathi A, De Souza CT. Multi-regulatory network of ROS: the interconnection of ROS, PGC-1 alpha, and AMPK-SIRT1 during exercise. Journal of physiology and biochemistry. 2017 Nov;73(4):487-94.[ 10.1007/s13105-017-0576-y ][PMID]
14. Nourolapour S, Abbassi Daloii A, Ziaolhagh SJ, Barari A. The effect of aerobic exercise and crocin consumption on the gene expression of BDNF, TrkB, dopamine, and serotonin in the cerebral cortex of rats induced with methamphetamine. Journal of Sport and Exercise Physiology. 2024 Jun 21;17(2):1-9.
15. Bahari H, Shahraki Jazinaki M, Aghakhani L, Amini MR, Noushzadeh Z, Khodashahi R, Malekahmadi M. Crocin Supplementation on Inflammation and Oxidative Stress: A Systematic Review and Meta‐Analysis. Phytotherapy Research. 2025 Jan;39(1):465-79. [][PMID]
16. Boussabbeh M, Haddar M, Sallem A, Chaieb A, Khdhiri R, Abid‐Essefi S, Mehdi M. Enhancing Male Fertility: The Role of Crocin in Boosting Sperm Motility Through Antioxidant Activity and Mitochondrial Pathways. Journal of Biochemical and Molecular Toxicology. 2025 May;39(5):e70275. [10.1002/ptr.8380 ][PMID]
17. Sánchez-Fernández C, Del Olmo-Aguado S, Artime E, Barros A, Fernández-Vega Cueto L, Merayo-Lloves J, Alcalde I. Immunocytochemical Analysis of Crocin against Oxidative Stress in Trigeminal Sensory Neurons Innervating the Cornea. Molecules. 2024 Jan 17;29(2):456. [10.3390/molecules29020456 ][PMID]
18. Xu E, Liu J, Liu H, Wang X, Xiong H. Inflammasome activation by methamphetamine potentiates lipopolysaccharide stimulation of IL-1β production in microglia. Journal of Neuroimmune Pharmacology. 2018 Jun;13(2):237-53. [10.1007/s11481-018-9780-y ][PMID]
19. Yang T, Zang S, Wang Y, Zhu Y, Jiang L, Chen X, Zhang X, Cheng J, Gao R, Xiao H, Wang J. Methamphetamine induced neuroinflammation in mouse brain and microglial cell line BV2: Roles of the TLR4/TRIF/Peli1 signaling axis. Toxicology letters. 2020 Oct 15;333:150-8. [10.1016/j.toxlet.2020.07.028 ][PMID]
20. He T, Han C, Liu C, Chen J, Yang H, Zheng L, Waddington JL, Zhen X. Dopamine D1 receptors mediate methamphetamine-induced dopaminergic damage: involvement of autophagy regulation via the AMPK/FOXO3A pathway. Psychopharmacology. 2022 Mar;239(3):951-64. [10.1007/s00213-022-06097-6 ][PMID]
21. Khaksari Z, Mehri F, Abdolvahab MH, Manavi MA, Nasab MH, Karbasi A, Baeeri M, Ranjbar A. Crocin and Nano-Crocin Mitigate Paraquat Hepatotoxicity by modulating expression of genes involved in oxidative stress and inflammation. Pharmaceutical nanotechnology. 2025. [10.2174/0122117385323941241211084423 ][PMID]
22. Su W, Wang Y, Shao S, Ye X. Crocin ameliorates neuroinflammation and cognitive impairment in mice with Alzheimer's disease by activating PI3K/AKT pathway. Brain and Behavior. 2024 May;14(5):e3503. [10.1002/brb3.3503 ][PMID]
23. Alizadehmoghaddam S, Pourabdolhossein F, Najafzadehvarzi H, Sarbishegi M, Saleki K, Nouri HR. Crocin attenuates the lipopolysaccharide-induced neuroinflammation via expression of AIM2 and NLRP1 inflammasome in an experimental model of Parkinson's disease. Heliyon. 2024 Feb 15;10(3). [10.1016/j.heliyon.2024.e25523 ][PMID]
24. Dastan M, Rajaei Z, Sharifi M, Salehi H. Crocin Improves Cognitive Impairment in LPS-treated Rats through Anti-Apoptotic, Anti-Inflammatory, and Antioxidant Activities. Molecular Neurobiology. 2025 May;62(5):5804-15. [10.1007/s12035-024-04638-y ][PMID]
25. Liu L, Tang J, Liang X, Li Y, Zhu P, Zhou M, Qin L, Deng Y, Li J, Wang Y, Jiang L. Running exercise alleviates hippocampal neuroinflammation and shifts the balance of microglial M1/M2 polarization through adiponectin/AdipoR1 pathway activation in mice exposed to chronic unpredictable stress. Molecular psychiatry. 2024 Jul;29(7):2031-42. [10.1038/s41380-024-02464-1 ][PMID]
26. Chen Y, Wu S. Effects of different exercise modalities on oxygenation stress, inflammatory factors, and drug craving in drug users during the audit period. Quality in Sport. 2025 May 11;41:60144.
27. Seo DY, Heo JW, Ko JR, Kwak HB. Exercise and neuroinflammation in health and disease. International neurourology journal. 2019 Nov 30;23(Suppl 2):S82. [10.5213/inj.1938214.107 ][PMID]
28. Lang X, Zhao N, He Q, Li X, Li X, Sun C, Zhang X. Treadmill exercise mitigates neuroinflammation and increases BDNF via activation of SIRT1 signaling in a mouse model of T2DM. Brain Research Bulletin. 2020 Dec 1;165:30-9. [10.1016/j.brainresbull.2020.09.015 ][PMID]
29. Spielman LJ, Little JP, Klegeris A. Physical activity and exercise attenuate neuroinflammation in neurological diseases. Brain research bulletin. 2016 Jul 1;125:19-29. [10.1016/j.brainresbull.2016.03.012 ][PMID]
30. Rakshe PS, Dutta BJ, Chib S, Maurya N, Singh S. Unveiling the interplay of AMPK/SIRT1/PGC-1α axis in brain health: Promising targets against aging and NDDs. Ageing research reviews. 2024 Apr 1;96:102255. [10.1016/j.arr.2024.102255 ][PMID]

  • Receive Date 10 March 2024
  • Revise Date 13 June 2025
  • Accept Date 29 June 2025