The effect of hydroalcoholic extract of Ziziphora clinopodioides L. on spatial memory and neuronal density of hippocampal CA1 region in rats with sporadic Alzheimer's disease

Document Type: Original Research Article


1 Department of Biology, Damghan Branch, Islamic Azad University, Damghan, Iran

2 Department of Biology, Mashhad Branch, Islamic Azad University, Mashhad, Iran.

3 Department of Biology, Damghan Branch, Islamic Azad University, Damghan, Iran.

4 Department of chemistry, Damghan Branch, Islamic Azad University, Damghan, Iran.


Objective: Alzheimer's disease is a neurodegenerative disorder associated with gradual loss of cognitive and memory abilities. It was shown that the hippocampus is one of the first structures in the brain that is affected by the disease. Ziziphora clinopodioides (Z. clinopodioides) is a member of Lamiaceae family and contains various substances.
Materials and Methods: In this experimental study, 72 adult male Wistar rats were used for behavioral and histopathologic studies. They were divided into nine groups included: control, negative control (Alzheimer), positive control (Alzheimer's treated with rivastigmine), aCSF (artificial cerebrospinal fluid) + ziziphora extract with doses of 200,400, and 600 mg/kg, and STZ (stereptozotocine)+ziziphoraextract in 200,400,600 mg/kg doses. The injury was created with bilaterally intraventricular injection. The spatial memory was studied by passive avoidance test and neuronal density was evaluated by dissector method. To examine the histopathological lesions, Congo red and toluidine blue staining were done. Data were analyzed by ANOVA Minitab software.
Results: The memory index (neuronal density and passive avoidance test results) showed a significant decrease in negative control group compared to control (p≤0.001). Treatment with the hydroalcoholic extract at the doses of 400 and 600 mg/kg showed a significant increase in memory index in rats with Alzheimer's disease (p≤0.001). The effect of 200 mg/kg extract was not significantly different from that of the negative control group. The results of histological analysis indicated beta-amyloid plaques formation in the control group as compared to the negative control group while treatment with the extract at the doses of 400 and 600 mg/kg, significantly reduced beta-amyloid plaques formation.  
Conclusion: These findings suggest that the extract of Z. clinopodioides can improve Alzheimer's condition and alleviate memory and histopathologic damages; also, it decreases beta-amyloid plaques and apoptosis in CA1 region of the hippocampus.


Main Subjects

Agrawal R, Tyagi E, Shukla R, Nath C. 2009. A study of brain insulin receptors, AChE activity and oxidative stress in rat model of ICV STZ induced dementia. Neuropharmacology, 56:779-787.

Aliev G, Obrenovich ME, Reddy VP. 2008. Antioxidant therapy in Alzheimer’s disease: theory and practice. Med Chem, 8:1395-1399

Babri S, Amani M, Mohaddes G, Alihemmati A, Ebrahimi H. 2012. Effect of aggregated β-amyloid (1-42) on synaptic plasticity of hippocampal dentate gyrus granule cells in vivo. Bio Impacts, BI 2:189-195.

Biasibetti R, dos Santos JPA, Rodrigues L. 2017 Hippocampal changes in STZ-model of Alzheimer’s disease are dependent on sex. Behav Brain Res, 316:205-214.

Butterfield DA, Drake J, Pocernich C, Castegna A. 2001. Evidence of oxidative damage in Alzheimer's disease brain: central role for amyloid β-peptide. Trends Mol Med, 7:548-554.

Carreiras M, Marco J.2004. Recent approaches to novel anti-Alzheimer therapy. Curr Pharm Des, 10:3167-3175.

Chen Y, Liang Z, Blanchard J. 2013. A non-transgenic mouse model (icv-STZ mouse) of Alzheimer’s disease: similarities to and differences from the transgenic model (3xTg-AD mouse). Mol Neurobiol, 47:711-725.

Cummings J, Winblad B. 2007. A rivastigmine patch for the treatment of Alzheimer’s disease and Parkinson’s disease dementia. Expert Rev Neurother, 7:1457-1463.

Dehghan Z, Sefidkon F, Khaniki G, Kalvandi R. 2010. Effects of some ecological factors on essential oil content and composition of Ziziphora clinopodioides Lam. subsp. rigida (Boiss.). Med Aromat Plants, 26:49-63.

Gill G, Woodward A, Casson I, Weston P.2009. Cardiac arrhythmia and nocturnal hypoglycaemia in type 1 diabetes—the ‘dead in bed’syndrome revisited. Diabetologia, 52:42-49.

Grünblatt E, SalkovicPetrisic M, Osmanovic J, Riederer P, Hoyer S. 2007. Brain insulin system dysfunction in streptozotocin intracerebroventricularly treated rats generates hyperphosphorylated tau protein. Neurochemistry, 101:757-770.

Hajiaghaee R, Akhondzadeh S. 2012. Herbal medicine in the treatment of Alzheimer’s disease. Med Plants, 1:1-7.

Hamann S. 2001. Cognitive and neural mechanisms of emotional memory. Trends Cognit Sci, 5:394-400.

Herring A, Ambrée O, Tomm M. 2009. Environmental enrichment enhances cellular plasticity in transgenic mice with Alzheimer-like pathology. Exp Neurol, 216:184-192.

Hoyer S, Lee SK, Löffler T, Schliebs R. 2000. Inhibition of the neuronal insulin receptor an in vivo model for sporadic Alzheimer disease? Ann N Y Acad Sci, 920:256-258.

Jain S, Shukla SD, Sharma K, Bhatnagar M. 2001. Neuroprotective effects of withania somnifera dunn in hippocampal subregions of female albino rat. Phytother Res, 15:544-548.

Janson J, Laedtke T, Parisi JE, O’Brien P, Petersen RC, Butler PC. 2004. Increased risk of type 2 diabetes in Alzheimer disease. Diabetes, 53:474-481.

Kazmierski J, Messini-Zachou C, Gkioka M, Tsolaki M. 2018. The impact of a long-term rivastigmine and donepezil treatment on all-cause mortality in patients with Alzheimer’s disease. Alzheimer's Dis. Other Dementias ®:1533317518775044.

Kennedy DO, Little W, Scholey A. B 2004. Attenuation of laboratory-induced stress in humans after acute administration of Melissa officinalis (Lemon Balm). Psychosom Med, 66:607-613.

Kim J, Huh J, Song S. 1996. The antioxidative mechanism of ursolic acid. Korean J Gerontol, 6:52-56.

Klyubin I, Walsh DM, Lemere CA.2005. Amyloid β protein immunotherapy neutralizes Aβ oligomers that disrupt synaptic plasticity in vivo. Nat Med, 11:556-560.

Kroner Z.2009.The relationship between Alzheimer's disease and diabetes: Type 3 diabetes? Altern Med, 14:373-378.

Kulišić T, Kriško A, Dragović-Uzelac V, Miloš M, Pifat G. 2007. The effects of essential oils and aqueous tea infusions of oregano (Origanum vulgare L. spp. hirtum), thyme (Thymus vulgaris L.) and wild thyme (Thymus serpyllum L.) on the copper-induced oxidation of human low-density lipoproteins. Food Sci Nutr, 58:87-93.

Kyari M.2008. Extraction and characterization of seed oils. Int Agrophys, 22:139-143.

Lane RM, Kivipelto M, Greig NH.2004. Acetylcholinesterase and its inhibition in Alzheimer disease. Clin Neuropharmacol, 27:141-149.

Lannert H, Hoyer S. 1998. Intracerebroventricular administration of streptozotocin causes long-term diminutions in learning and memory abilities and in cerebral energy metabolism in adult rats. Behav Neurosci, 112:1199-1204.

Li G, Ma R, Huang C. 2008. Protective effect of erythropoietin on β-amyloid-induced PC12 cell death through antioxidant mechanisms. Neurosci Lett, 442:143-147.

Mahdy K, Shaker O, Wafay H, Nassar Y, Hassan H, Hussein A. 2012. Effect of some medicinal plant extracts on the oxidative stress status in Alzheimer’s disease induced in rats. Eur Rev Med Pharmacol Sci, 16:31-42.

Mehla J, Pahuja M, Gupta YK. 2013. Streptozotocin-induced sporadic Alzheimer's disease: selection of appropriate dose. Alzheimer's Dis, 33:17-21.

Miu AC, Andreescu CE, Vasiu R, Olteanu AI. 2003. A behavioral and histological study of the effects of long-term exposure of adult rats to aluminum. Neuroscience, 113:1197-1211.

Moazedi A, Ghotbedin Z, Parham G. 2015. The effect of Zncl2 on passive avoidance and motor activity learing due to administration of Alcl3 in male rats. Physiol Pharmacol, 1386:146-152.

Naghizadeh B, Mansouri M, Ghorbanzadeh B, Farbood Y, Sarkaki A. 2013. Protective effects of oral crocin against intracerebroventricular streptozotocin-induced spatial memory deficit and oxidative stress in rats. Phytomedicine, 20:537-542.

Nezhadali A, Shirvan BZ. 2010. Separation, Identification and Determination ofVolatile Compounds of Ziziphora persica BungeUsing HS-SPME/GC-MS. Int J Environ Sci Dev, 1:115-119.

Nilsson MR. 2004. Techniques to study amyloid fibril formation in vitro. Methods, 34:151-160.

Paxinos G, Watson C. 1986. The rat in stereotaxic coordinates. J Anat, 191:315-317.

Palazzi X, Bordier N. 2008. The marmoset brain in stereotaxic coordinates. In:Palazzi, X, The Marmoset Brain in Stereotaxic Coordinates, pp. 51-59, New York,Springer.

Perry EK, Pickering AT, Wang WW, Houghton P, Perry NS.1998. Medicinal plants and Alzheimer's disease: Integrating ethnobotanical and contemporary scientific evidence. Complementary Altern Med, 4:419-428.

Perry N, Menzies R, Hodgson F. 2018. A randomised double-blind placebo-controlled pilot trial of a combined extract of sage, rosemary and melissa, traditional herbal medicines, on the enhancement of memory in normal healthy subjects, including influence of age. Phytomedicine, 39:42-48.

Perry NS, Houghton PJ, Theobald A, Jenner P, Perry EK.2000. Invitro inhibition of human erythrocyte acetylcholinesterase by Salvia lavandulaefolia essential oil and constituent terpenes. Am J Pharm Pharmacol, 52:895-902.

Rabiei Z, Mokhtari S, Asgharzade S, Gholami M, Rahnama S, Rafieian-kopaei M. 2015. Inhibitory effect of Thymus vulgaris extract on memory impairment induced by scopolamine in rat. Asian Pac J Trop Biomed, 5:845-851.

Reisi P, Alaei H, Babri S, Sharifi MR, Mohaddes G. 2009. Effects of treadmill running on spatial learning and memory in streptozotocin-induced diabetic rats. Neurosci Lett, 455:79-83.

Rice-Evans CA, Miller NJ, Paganga G. 1996. Structure-antioxidant activity relationships of flavonoids and phenolic acids. Adv. Free Radical Biol. Med, 20:933-956.

Roig y Mesa JT. 1974. Plantas medicinales, aromáticas o venenosas de Cuba. In:Roig y,Plantas medicinales, pp. 120-122, Cuba, informed.

Rowan M, Klyubin I, Wang Q, Hu N, Anwyl R. 2007. Synaptic memory mechanisms: Alzheimer's disease amyloid β-peptide-induced dysfunction. Portland Press Limited, 4:1219-1223.

SalkovicPetrisic M, Tribl F, Schmidt M, Hoyer S, Riederer P. 2006. Alzheimerlike changes in protein kinase B and glycogen synthase kinase3 in rat frontal cortex and hippocampus after damage to the insulin signalling pathway. J Neurochem, 96:1005-1015.

Singh B, Sharma B, Jaggi AS, Singh N. 2013. Attenuating effect of lisinopril and telmisartan in intracerebroventricular streptozotocin induced experimental dementia of Alzheimer’s disease type: possible involvement of PPAR-γ agonistic property. J Renin Angiotensin Aldosterone Syst, 14:124-136.

Squire LR, Zola SM. 1996. Structure and function of declarative and nondeclarative memory systems. Proc. Natl Acad Sci, 93:13515-13522.

Sul D, Kim H-S, Lee D, Joo SS, Hwang KW, Park S-Y. 2009. Protective effect of caffeic acid against beta-amyloid-induced neurotoxicity by the inhibition of calcium influx and tau phosphorylation. Life Sci, 84:257-262.

Tota S, Kamat PK, Shukla R, Nath C. 2011. Improvement of brain energy metabolism and cholinergic functions contributes to the beneficial effects of silibinin against streptozotocin induced memory impairment. Beha. Brain Res, 221:207-215.

Williams RJ, Spencer JP.2012. Flavonoids, cognition, and dementia: actions, mechanisms, and potential therapeutic utility for Alzheimer disease. Adv Free Radical Biol Med, 52:35-45.

Xiao J, Tundis R.2013. Natural products for Alzheimer's disease therapy: basic and application. J Pharm Pharmacol, 65:1679-1680.

Xing S, Zhang P, Ji Q, Jia H, Wang X.2010. Essential oil compositions and antioxidant activities of two Ziziphora species in Xinjiang. Food Sci, 31:154-159.

Yang LP, Keating GM.2007. Rivastigmine Transdermal Patch. CNS drugs, 21:957-965.

Zhou L, Tan S, Shan Y-l.2016. Baicalein improves behavioral dysfunction induced by Alzheimer’s disease in rats. Neuropsychiatr Dis Treat, 12:3145-3149