Protective effects of Vitex agnus-castus in ovariectomy mice following permanent middle cerebral artery occlusion

Document Type: Short Communication


1 Student Research Committee, Rafsanjan University of Medical Sciences, Rafsanjan, Iran

2 Research Center for Tropical and Infectious Diseases, Kerman University of Medical Sciences, Kerman, Iran

3 Physiology-Pharmacology Research Center, Research Institute of Basic Medical Sciences, Rafsanjan University of Medical Sciences, Rafsanjan, Iran

4 Department of Physiology and Pharmacology, School of Medicine, Rafsanjan University of Medical Sciences, Rafsanjan, Iran


Objective(s): Previous studies have indicated that phytoestrogens induce estrogenic as well as anti-inflammatory effects, and they are found in high abundance in the extracts of some herbs such as Vitex Agnus Castus (VAC). Therefore, we investigated the effect of VAC extract on ovariectomized mice after the induction of permanent middle cerebral artery occlusion (PMCAO) model.
Materials and Methods: In this study, 50 mice ranging from 25 to 35 g were divided into five experimental groups as follows: Control, VAC, Estrogen, Tamoxifen, and Tamoxifen-VAC.  Animals were ovariectomized, and after 30 days of treatment, they were given PMCAO induction. Behavioral assessment (adhesive removal and wire hanging tests) was evaluated 24 hr, 48 hr, and one week after induction of stroke. The infarct volume, as well as serum levels of matrix metalloproteinase-9 (MMP-9) and interleukin-10 (IL-10), were measured one week after stroke.
Results: One week after stroke, in both VAC and estrogen groups, the infarct size reduced in comparison with the control group. Estrogen and VAC extract improved adhesive removal and wire hanging test, increased the level of IL-10, and decreased the level of MMP-9 compared with the control group. In addition, co-administration of tamoxifen and VCA extract had no significant effect on measured indices compared with control and tamoxifen groups.
Conclusion: Based on our findings, VAC extract has neuroprotective properties and can reduce stroke injuries in PMCAO-induced ovariectomized mice via anti-inflammatory and estrogenic properties.


Main Subjects

1. Prongay KD, Lewis AD, Hurn PD, Murphy SJ. Dietary soy may not confound acute experimental stroke infarct volume outcomes in ovariectomized female rats. Lab Anim 2010; 44:238-246.
2. Liu R, Wen Y, Perez E, Wang X, Day AL, Simpkins JW, et al. 17β-Estradiol attenuates blood–brain barrier disruption induced by cerebral ischemia–reperfusion injury in female rats. Brain Res 2005; 1060:55-61.
3.  Merchenthaler I, Dellovade TL, Shughrue PJ. Neuroprotection by estrogen in animal models of global and focal ischemia. Ann N Y Acad Sci 2003; 1007:89-100.
4. Habib P, Dreymueller D, Ludwig A, Beyer C, Dang J. Sex steroid hormone-mediated functional regulation of microglia-like BV-2 cells during hypoxia. J Steroid Biochem Mol Biol 2013; 138:195-205.
5. Zhang J, Lapato A, Bodhankar S, Vandenbark AA, Offner H. Treatment with IL-10 producing B cells in combination with E2 ameliorates EAE severity and decreases CNS inflammation in B cell-deficient mice. Metab Brain Dis 2015; 30:1117-1127.
6. Protti GG, Gagliardi RJ, Forte WC, Sprovieri SR. Interleukin-10 may protect against progressing injury during the acute phase of ischemic stroke. Arq Neuropsiquiatr 2013; 71:846-851.
7. Viscoli CM, Brass LM, Kernan WN, Sarrel PM, Suissa S, Horwitz RI. A clinical trial of estrogen-replacement therapy after ischemic stroke. N Engl J Med 2001; 345:1243-1249.
8. Duncan AM, Phipps WR, Kurzer MS. Phyto-oestrogens. Best Pract Res Clin Endocrinol Metab 2003; 17:253-271.
9. Sirotkin AV, Harrath AH. Phytoestrogens and their effects. Eur J Pharmacol 2014; 741:230-236.
10. Yaman M EO, Cosar M, Bas O, Sahin O, Mollaoglu H, et al. Oral administration of avocado soybean unsaponifiables (ASU) reduces ischemic damage in the rat hippocampus. Arch Med Res 2007; 38:489-494.
11. Castelló-Ruiz M TG, Burguete M, Salom J, Gil J, Miranda F, et al. Soy-derived phytoestrogens as preventive and acute neuroprotectors in experimental ischemic stroke: influence of rat strain. Phytomedicine 2011; 18:513-515.
12. Soni M, Rahardjo TB, Soekardi R, Sulistyowati Y, Lestariningsih, Yesufu-Udechuku A, et al. Phytoestrogens and cognitive function: a review. Maturitas 2014; 77:209-220.
13. Aras AB, Guven M, Akman T, Alacam H, Kalkan Y, Silan C, et al. Genistein exerts neuroprotective effect on focal cerebral ischemia injury in rats. Inflammation 2015; 38:1311-1321.
14. E Orhan I, F Nabavi S, Daglia M, C Tenore G, Mansouri K, M Nabavi S. Naringenin and atherosclerosis: A review of literature. Curr Pharm Biotechnol 2015; 16:245-251.
15. Kashtriya R, Shaikh Y, Nazeruddin G. A Brief Review: Flavonoids as a pharmacophore. J Applicable Chem 2015; 4:801-817.
16. Schreihofer DA, Deutsch C, Lovekamp-Swan T, Sullivan JC, Dorrance AM. Effect of high soy diet on the cerebrovasculature and endothelial nitric oxide synthase in the ovariectomized rat. Vascul Pharmacol 2010; 52:236-242.
17. Rani A, Sharma A. The genus Vitex: A review. Pharmacognosy reviews 2013; 7:188-197.
18. Marotta F, Mao G, Liu T, Chui D, Lorenzetti A, Xiao Y, et al. Anti‐inflammatory and neuroprotective effect of a phytoestrogen compound on rat microglia. Ann N Y Acad Sci 2006; 1089:276-281.
19. Ghannadi A, Bagherinejad M, Abedi D, Jalali M, Absalan B, Sadeghi N. Antibacterial activity and composition of essential oils from Pelargonium graveolens L’Her and Vitex agnus-castus L. Iran J Microbiol 2012; 4:171-176.
20. Khalilzadeh E, Vafaei Saiah G, Hasannejad H, Ghaderi A, Ghaderi S, Hamidian G, et al. Antinociceptive effects, acute toxicity and chemical composition of Vitex agnus-castus essential oil. Avicenna J Phytomed 2015; 5:218-230.
21. Liu J, Burdette J, Sun Y, Deng S, Schlecht S, Zheng W, et al. Isolation of linoleic acid as an estrogenic compound from the fruits of Vitex agnus-castus L.(chaste-berry). Phytomedicine 2004; 11:18-23.
22. Chen SN, Friesen JB, Webster D, Nikolic D, van Breemen RB, Wang ZJ, et al. Phytoconstituents from Vitex agnus-castus fruits. Fitoterapia. 2011; 82:528-533.
23. Honari N, Pourabolli I, Hakimizadeh E, Roohbakhsh A, Shamsizadeh A, Vazirinejad R, et al. Effect of Vitex agnus-castus extraction on anxiety-like behaviors in ovariectomized rats. J Babol Univ Med Sci 2012; 14:29-35.
24. Fatemi I, Heydari S, Kaeidi A, Shamsizadeh A, Hakimizadeh E, Khaluoi A, et al. Metformin ameliorates the age-related changes of d-galactose administration in ovariectomized mice. Fundam Clin Pharmacol. 2018; 32:392-399.
25. Fatemi I, Delrobaee F, Bahmani M, Shamsizadeh A, Allahtavakoli M. The effect of the anti-diabetic drug metformin on behavioral manifestations associated with ovariectomy in mice. Neurosci Lett 2019; 690:95-98.
26. Delrobaei F, Fatemi I, Shamsizadeh A, Allahtavakoli M. Ascorbic acid attenuates cognitive impairment and brain oxidative stress in ovariectomized mice. Pharmacol Rep 2018; 71:133-138.
27. Sharma K, Mehra RD. Long-term administration of estrogen or tamoxifen to ovariectomized rats affords neuroprotection to hippocampal neurons by modulating the expression of Bcl-2 and Bax. Brain Res 2008; 1204:1-15.
28. Allahtavakoli M, Amin F, Esmaeeli‐Nadimi A, Shamsizadeh A, Kazemi‐Arababadi M, Kennedy D. Ascorbic acid reduces the adverse effects of delayed administration of tissue plasminogen activator in a rat stroke Model. Basic Clin Pharmacol Toxicol 2015; 117:335-339.
29. Hadadianpour Z, Fatehi F, Ayoobi F, Kaeidi A, Shamsizadeh A, Fatemi I. The effect of orexin-A on motor and cognitive functions in a rat model of Parkinson’s disease. Neurol Res 2017; 39:845-851.
30. Hassanshahi A, Shafeie SA, Fatemi I, Hassanshahi E, Allahtavakoli M, Shabani M, et al. The effect of Wi-Fi electromagnetic waves in unimodal and multimodal object recognition tasks in male rats. Neurol Sci 2017; 38:1069-1076.
31. Hakimizadeh E, Shamsizadeh A, Roohbakhsh A, Arababadi MK, Hajizadeh MR, Shariati M, et al. TRPV1 receptor-mediated expression of toll-like receptors 2 and 4 following permanent middle cerebral artery occlusion in rats. Iran J Basic Med Sci 2017; 20:863-869.
32. Schreihofer DA, Do KD, Schreihofer AM. High-soy diet decreases infarct size after permanent middle cerebral artery occlusion in female rats. Am J Physiol Regul Integr Comp Physiol 2005; 289:103-108.
33. Wang W, Tang L, Li Y, Wang Y. Biochanin A protects against focal cerebral ischemia/reperfusion in rats via inhibition of p38-mediated inflammatory responses. J Neurol Sci 2015; 348:121-125.
34. Huang G, Cao X, Zhang X, Chang H, Yang Y, DU W, et al. Effects of soybean isoflavone on the notch signal pathway of the brain in rats with cerebral ischemia. J Nutr Sci Vitaminol 2009; 55:326-331.
35. Dong X, Song Y-N, Liu W-G, Guo X-L. Mmp-9, a potential target for cerebral ischemic treatment. Current neuropharmacology 2009; 7:269-273.
36. Rachoń D, Rimoldi G, Wuttke W. In vitro effects of genistein and resveratrol on the production of interferon-γ (IFN-γ) and interleukin-10 (IL-10) by stimulated murine splenocytes. Phytomedicine 2006; 13:419-424.