Neuroprotective effects of ROCK inhibition on hippocampal energy metabolism in a rat model of metabolic syndrome

Articles in Press

Document Type : Original Article

Authors

1 Department of Biology, Faculty of Exact and Natural Sciences, Iv. Javakhishvili Tbilisi State University, Tbilisi, Georgia

2 Faculty of Medicine, Iv. Javakhishvili Tbilisi State University, Tbilisi, Georgian

3 Department of Pharmacology, Faculty of Medicine, Iv. Javakhishvili Tbilisi State University, Tbilisi, Georgia

10.22038/ijbms.2026.93965.20252

Abstract

Objective(s): Metabolic syndrome (MS) is associated with insulin resistance, hyperglycemia, and dyslipidemia, leading to impaired neuronal energy metabolism and hippocampal dysfunction. Rho-associated kinase (ROCK) has been implicated in metabolic and neuroinflammatory dysregulation; however, its role in MS-induced hippocampal energy imbalance remains unclear. This study aimed to determine whether pharmacological ROCK inhibition with fasudil restores hippocampal energy metabolism in a rat model of fructose-induced MS.
Materials and Methods: Male Lewis rats were divided into control, MS, and MS+fasudil groups. Plasma and hippocampal metabolic parameters were assessed using biochemical assays. Activities of glycolytic enzymes, Krebs cycle enzymes, and mitochondrial respiratory chain complexes I–IV were measured. Phosphatidylinositol 3-kinase/Protein kinase B/Mechanistic target of rapamycin (PI3K/AKT/mTOR) pathway components, including phosphorylated PI3K, AKT, mTOR, Phosphatase and tensin homolog (PTEN), and Ras homolog enriched in brain (Rheb), were analyzed by Western blotting.
Results: MS disrupted glucose homeostasis and significantly suppressed the activities of glycolytic, mitochondrial, and respiratory chain enzymes in the hippocampus. These changes were accompanied by reduced phosphorylation of PI3K, AKT, and mTOR, increased PTEN expression, and decreased Rheb levels, indicating impaired PI3K/AKT/mTOR signaling. Fasudil treatment normalized plasma and hippocampal glucose levels, restored enzymatic activities across major energy-producing pathways, and reactivated PI3K/AKT/mTOR signaling by reducing PTEN and restoring AKT and Rheb activity.
Conclusion: These findings demonstrate that ROCK overactivation contributes to central insulin resistance and hippocampal energy dysfunction in MS. ROCK inhibition with fasudil effectively reverses these alterations, highlighting ROCK as a potential therapeutic target for preserving hippocampal metabolic function in MS.

Keywords

Main Subjects

References

1. Neeland IJ, Lim S, Tchernof A, Gastaldelli A, Rangaswami J, Ndumele CE, et al. Metabolic syndrome. Nat Rev Dis Primers 2024; 10: 77.
2. Eckel RH, Grundy SM, Zimmet PZ. The metabolic syndrome. Lancet 2005; 365: 1415–1428.
3. Rochlani Y, Pothineni NV, Kovelamudi S, Mehta JL. Metabolic syndrome: Pathophysiology, management, and modulation by natural compounds. Ther Adv Cardiovasc Dis 2017; 11: 215-225.
4. Fahed G, Aoun L, Bou Zerdan M, Allam S, Bou Zerdan M, Bouferraa Y, et al. Metabolic syndrome: Updates on pathophysiology and management in 2021. Int J Mol Sci 2022; 23: 786.
5. Swarup S, Ahmed I, Grigorova Y, Zeltser R. Metabolic syndrome. In: StatPearls. Treasure Island (FL): StatPearls Publishing; 2025.
6. Noubiap JJ, Nansseu JR, Lontchi-Yimagou E, Nkeck JR, Nyaga UF, Ngouo AT, et al. Global, regional, and country estimates of metabolic syndrome burden in children and adolescents in 2020: A systematic review and modelling analysis. Lancet Child Adolesc Health 2022; 6: 158-170.
7. Zouridis S, Nasir AB, Aspichueta P, Syn WK. The link between metabolic syndrome and the brain. Digestion 2025; 106: 203-211.
8. Tahmi M, Palta P, Luchsinger JA. Metabolic syndrome and cognitive function. Curr Cardiol Rep 2021; 23: 180.
9. Zuin M, Roncon L, Passaro A, Cervellati C, Zuliani G. Metabolic syndrome and the risk of late onset Alzheimer’s disease: an updated review and meta-analysis. Nutr Metab Cardiovasc Dis 2021; 31: 2244–2252.
10. Al-Kuraishy HM, Jabir MS, Albuhadily AK, Al-Gareeb AI, Rafeeq MF. The link between metabolic syndrome and Alzheimer disease: a mutual relationship and long rigorous investigation. Ageing Res Rev 2023; 91: 102084.
11. Ezkurdia M, Ramírez MJ, Solas M. Metabolic syndrome as a risk factor for Alzheimer’s disease: A focus on insulin resistance. Int J Mol Sci 2023; 24: 4354.
12. Gómez-Apo E, Mondragón-Maya A, Ferrari-Díaz M, Silva-Pereyra J. Structural brain changes associated with overweight and obesity. J Obes 2021; 2021: 6613385.
13. Fuentes E, Venegas B, Muñoz-Arenas G, Moran C, Vazquez-Roque RA, et al. High-carbohydrate and fat diet consumption causes metabolic deterioration, neuronal damage, and loss of recognition memory in rats. J Chem Neuroanat 2023; 129: 102237.
14. de la Monte SM. Insulin resistance and neurodegeneration: Progress towards the development of new therapeutics for Alzheimer’s disease. Drugs 2017; 77: 47-65.
15. Kouvari M, D’Cunha NM, Travica N, Sergi D, Zec M, Marx W, et al. Metabolic syndrome, cognitive impairment and the role of diet: A narrative review. Nutrients 2022; 14: 333.
16. Chen W, Cai W, Hoover B, Kahn CR. Insulin action in the brain: Cell types, circuits, and diseases. Trends Neurosci 2022; 45: 384–400.
17. Tan BL, Norhaizan ME. Effect of high-fat diets on oxidative stress, cellular inflammatory response and cognitive function. Nutrients 2019; 11: 2579.
18. Candelario-Jalil E, Dijkhuizen RM, Magnus T. Neuroinflammation, stroke, blood-brain barrier dysfunction, and imaging modalities. Stroke 2022; 53: 1473–1486.
19.  Grillo CA, Woodruff JL, Macht VA, Reagan LP. Insulin resistance and hippocampal dysfunction: Disentangling peripheral and brain causes from consequences. Exp Neurol 2019; 318: 71-77. 
20.  Spinelli M, Fusco S, Grassi C. Brain insulin resistance impairs hippocampal plasticity. Vitam Horm 2020; 114: 281-306. 
21.  Erichsen JM, Woodruff JL, Grillo CA, Piroli GG, Fadel JR, Reagan LP. Hippocampal-specific insulin resistance elicits synaptic effects on glutamate neurotransmission. J Neurochem 2025; 169:  e70083.
22. Spinelli M, Fusco S, Grassi C. Brain insulin resistance and hippocampal plasticity: Mechanisms and biomarkers of cognitive decline. Fron Neurosci 2019; 13: 788. 
23.  He Y, Sun M, Qu M, Lu Y, Yang H, Wang R, et al. Brain insulin signaling pathway regulation of hippocampal neuroplasticity in neurocognitive disorders: Mechanisms and therapeutic implications. J Integr Neurosci 2025; 24: 39446. 
24. Battú CE, Rieger D, Loureiro S, Furtado GV, Bock H, Saraiva-Pereira ML, et al. Alterations of PI3K and Akt signaling pathways in the hippocampus and hypothalamus of Wistar rats treated with highly palatable food. Nutr Neurosci 2012; 15: 10-17.
25. Yamamoto T, Ugawa Y, Kawamura M, Yamashiro K, Kochi S, Ideguchi H, et al. Modulation of microenvironment for controlling the fate of periodontal ligament cells: The role of Rho/ROCK signaling and cytoskeletal dynamics. J Cell Commun Signal 2018; 12: 369-378.
26. Dai C, Khalil RA. Calcium signaling dynamics in vascular cells and their dysregulation in vascular disease. Biomolecules 2025; 15: 892.
27. Koch JC, Tatenhorst L, Roser AE, Saal KA, Tönges L, Lingor P. ROCK inhibition in models of neurodegeneration and its potential for clinical translation. Pharmacol Ther 2018; 189: 1-21.
28. Chong CM, Ai N, Lee SM. ROCK in CNS: Different roles of isoforms and therapeutic target for neurodegenerative disorders. Curr Drug Targets 2017; 18: 455-462.
29. Iyer M, Subramaniam MD, Venkatesan D, Cho SG, Ryding M, Meyer M, et al. Role of RhoA-ROCK signaling in Parkinson’s disease. Eur J Pharmacol 2021; 894: 173815.
30. Schmidt SI, Blaabjerg M, Freude K, Meyer M. RhoA signaling in neurodegenerative diseases. Cells 2022; 11: 1520.
31. Lu W, Chen Z, Wen J. The role of RhoA/ROCK pathway in the ischemic stroke-induced neuroinflammation. Biomed Pharmacother 2023; 165: 115141.
32. Zhu YT, Zhang Q, Xie HY, Yu KW, Xu GJ, Li SY, et al. Environmental enrichment combined with fasudil promotes motor function recovery and axonal regeneration after stroke. Neural Regen Res 2021; 16: 2512-2520.
33. Muraoka S, Izumi T, Nishida K, Chrétien B, Ishii K, Takeuchi I, et al. RECOVER study: A multicenter retrospective cohort study and comparison of the efficacy and safety of clazosentan and fasudil in patients with aneurysmal subarachnoid hemorrhage. J Neurosurg 2025; 143: 624–633.
34. Tatenhorst L, Eckermann K, Dambeck V, Fonseca-Ornelas L, Walle H, Lopes da Fonseca T, et al. Fasudil attenuates aggregation of α-synuclein in models of Parkinson’s disease. Acta Neuropathol Commun 2016; 4: 39.
35. Shibuya M, Hirai S, Seto M, Satoh S, Ohtomo E. Effects of fasudil in acute ischemic stroke: results of a prospective placebo-controlled double-blind trial. J Neurol Sci 2005; 238: 31-39.
36. Dong J, Wang Z, Li L, Zhang M, Wang S, Luo Y, et al. Fasudil alleviates postoperative neurocognitive disorders in mice by downregulating the surface expression of α5GABAAR in hippocampus. CNS Neurosci Ther 2024; 30: e70098.
37. National Research Council (US) Committee for the Update of the Guide for the Care. Guide for the care and use of laboratory animals. 8th ed. Washington, DC: National Academies Press; 2011.
38. American Veterinary Medical Association. AVMA guidelines for the euthanasia of animals. 2020 ed. Schaumburg; 2020.
39. Paxinos G, Watson C. The Rat Brain in Stereotaxic Coordinates. 7th ed. Academic Press; 2014.
40. Zhgenti N, Bibilashvili O, Shengelia M, Burjanadze G, Koshoridze M, Davitashvili E, et al. Effect of nicotine on the energy metabolism of substantia nigra cells in MPTP-induced Parkinson’s disease. Iran J Basic Med Sci 2025; 28: 1417-1427.
41. Zhgenti N, Bibilashvili O, Burjanadze G, Shengelia M, Koshoridze M, Davitashvili E, et al. Dietary plant-derived lectins induce oxidative stress, metabolic dysfunction, apoptosis and neuroinflammation in mice brain. Nutr Neurosci 2025; 28: 1386–1398.
42. Shanshiashvili L, Tsitsilashvili E, Dabrundashvili N, Kalandadze I, Mikeladze D. Metabotropic glutamate receptor 5 may be involved in macrophage plasticity. Biol Res 2017; 50: 4-13.
43. Zhgenti N, Bakuradze E, Bibilashvili O, Koshoridze N. The role of BDNF/PI3K/AKT/Nrf2 signaling in nicotine’s protective effects against MPTP-induced Parkinson’s disease. Neurosci Lett 2025; 868: 138412.
44. Lubawy M, Formanowicz D. High-fructose diet-induced hyperuricemia accompanying metabolic syndrome-mechanisms and dietary therapy proposals. Int J Environ Res Public Health 2023; 20: 3596.
45. Pan Y, Kong LD. High fructose diet-induced metabolic syndrome: Pathophysiological mechanism and treatment by traditional Chinese medicine. Pharmacol Res 2018; 130: 438-450.
46. Hannou SA, Haslam DE, McKeown NM, Herman MA. Fructose metabolism and metabolic disease. J Clin Invest 2018; 128: 545–555.
47. Softic S, Cohen DE, Kahn CR. Role of dietary fructose and hepatic de novo lipogenesis in fatty liver disease. Dig Dis Sci 2016; 61: 1282–1293.
48. Jung S, Bae H, Song WS, Jang C. Dietary fructose and fructose-induced pathologies. Annu Rev Nutr 2022; 42: 45-66.
49. García-Berumen CI, Ortiz-Avila O, Vargas-Vargas MA, Del Rosario-Tamayo BA, Guajardo-López C, Saavedra-Molina A, et al. The severity of rat liver injury by fructose and high fat depends on the degree of respiratory dysfunction and oxidative stress induced in mitochondria. Lipids Health Dis 2019; 18: 78.
50. Cioffi F, Senese R, Lasala P, Ziello A, Mazzoli A, Crescenzo R, et al. Fructose-rich diet affects mitochondrial DNA damage and repair in rats. Nutrients 2017; 9: 323.
51. Zhang X, Zhang JH, Chen XY, Hu QH, Wang MX, Jin R, et al. Reactive oxygen species-induced TXNIP drives fructose-mediated hepatic inflammation and lipid accumulation through NLRP3 inflammasome activation. Antioxid Redox Signal 2015; 22: 848-870.
52. Huang H, Lee DH, Zabolotny JM, Kim YB. Metabolic actions of Rho-kinase in periphery and brain. Trends Endocrinol Metab 2013; 24: 506-514.
53. Sawada N, Liao JK. Rho/Rho-associated coiled-coil forming kinase pathway as therapeutic targets for statins in atherosclerosis. Antioxid Redox Signal 2014; 20: 1251-1267. 
54. Huang H, Lee DH, Zabolotny JM, Kim YB. Metabolic actions of Rho-kinase in periphery and brain. Trends Endocrinol Metab 2013; 24: 506-514.
55. Wei L, Shi J. Insight into rho kinase isoforms in obesity and energy homeostasis. Front Endocrinol 2022; 13: 886534.
56. Li G, Liu L, Shan C, Cheng Q, Budhraja A, Zhou T, et al. RhoA/ROCK/PTEN signaling is involved in AT-101-mediated apoptosis in human leukemia cells in vitro and in vivo. Cell Death Dis 2014; 5: e998.
57. Walker CL, Liu NK, Xu XM. PTEN/PI3K and MAPK signaling in protection and pathology following CNS injuries. Front Biol 2013; 8: 10.
Iranian Journal of Basic Medical Sciences

Articles in Press, Accepted Manuscript
Available Online from 20 June 2026

Files

Share

How to cite

Statistics