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Journal of Integrative Neuroscience  2018, Vol. 17 Issue (1): 19-25    DOI: 10.31083/JIN-170035
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The role of rosemary extract in degeneration of hippocampal neurons induced by kainic acid in the rat: A behavioral and histochemical approach
Elahe Naderali1, Farnaz Nikbakht2, Sattar Norouzi Ofogh3, Homa Rasoolijazi1, 4, *()
1 Anatomy Department, Medical school, Iran University of Medical Sciences, Tehran, Iran
2 Physiology Department, Medical School, Iran University of Medical Sciences, Tehran, Iran
3 Neuroscience Department, School of Advanced Technologies in Medicine, Iran University of Medical Sciences, Tehran, Iran
4 Cellular and Molecular Research Center, Iran University of Medical Sciences, Tehran, Iran
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Systemic kainic acid administration has been used to induce experimental temporal lobe epilepsy in rats. The aim was to evaluate the neuroprotective effect of rosemary extract (40% Carnosic acid) against kainic acid-induced neurotoxicity in hippocampus and impaired learning and memory. Subjects received a single dose of kainic acid (9.5 mg/kg) intraperitoneally, were observed for two hours, and scored from 0 (for normal, no convulsion) to five (for continuous generalized limbic seizures). Rosemary extract (100 mg/kg, orally) was administered daily for 23 days, starting a week before kainic acid injection. Neuronal degeneration in hippocampus was demonstrated by using Fluoro-Jade B immunofluorescence. The number of pyramidal cells in hippocampus was evaluated by Nissl staining. Also, the Morris water maze and shuttle box were used to assess spatial memory and passive avoidance learning, respectively. Results revealed that, after treatment with rosemary extract, neuronal loss in CA1 decreased significantly in subjects in the kainic acid + rosemary extract group. Morris water navigation task results revealed that spatial memory impairment decreased in subjects in the kainic acid + rosemary extract group. Furthermore, results in shuttle box testing showed that passive avoidance learning impairment significantly improved for subjects in the kainic acid + rosemary extract group. These results suggest that rosemary extract improves spatial and working memory deficits and also, due to its antioxidant activities, neuronal degeneration induced by the toxicity of kainic acid in rat hippocampus.

Key words:  Epilepsy      kainic acid      neurotoxicity      rosemary      learning and memory      rosemary extract     
Submitted:  04 February 2017      Accepted:  24 May 2017      Published:  15 February 2018     
*Corresponding Author(s):  Homa Rasoolijazi     E-mail:

Cite this article: 

Elahe Naderali, Farnaz Nikbakht, Sattar Norouzi Ofogh, Homa Rasoolijazi. The role of rosemary extract in degeneration of hippocampal neurons induced by kainic acid in the rat: A behavioral and histochemical approach. Journal of Integrative Neuroscience, 2018, 17(1): 19-25.

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Fig.1.  Effect of KA (kainic acid) with or without RE (rosemary extract) on occurrence and intensity of seizure. Subjects in the control and RE groups received vehicle and rosemary extract, respectively. There was no sign (score 0) of the occurrence of seizure in these subjects. Other groups received KA alone (KA group) or with RE (KA+RE group) and were evaluated for signs of seizure. **p < 0.01 compared with the KA+RE group.

Fig.2.  Effects of systemic administration of KA alone or in combination with RE administration on memory retrieval in the passive avoidance task. On the test day, step-through latency for subjects was assessed as a measure of passive avoidance learning. ***p < 0.001 compared with the control and RE group, **p < 0.01 compared with the KA+RE group.

Fig.3.  Effects of systemic administration of KA alone or in combination with RE on spatial memory formation in the Morris water maze test. Animals were classified into four groups according to the administration (Control, RE, KA or KA+RE) and spatial memory formation was evaluated over 5 days. On days 1 and 5, there were no significant differences among groups in Morris water maze performance. On days 2 and 3, KA administration impaired spatial learning in the Morris water maze and caused a significant increase in escape latency when compared to both the control and RE groups. On day 2, post hoc analysis also revealed that escape latency decreased (not significantly) for the KA+RE group compared to the KA group (p = 0.06). On day 4, significant differences were observed between the KA group and the control group but not with the RE group. **p < 0.01 compared to the RE group, *p < 0.05 compared to the control and RE groups.

Fig.4.  The spatial probe test measured the time spent in the target quadrant (TQ). KA (kainic acid) administration caused subjects in the KA group to spend less time in the TQ compare to subjects in the control group. This performance by subjects demonstrated a spatial memory disability induced by KA. *p< 0.05 compared to control groups.

Fig.5.  Improvement effects of RE on hippocampal neurons survival influenced by KA (kainic acid). KA administration significantly decreased the number of intact neurons in CA1 cell layer. RE administration alone had no effect on neuronal survive in this layer compared to the control group, but RE reversed the KA-neurotoxicity in CA1 in the KA+RE group. ***p < 0.001 compared with the control group, ###p < 0.001 compared with the KA group.

Fig.6  Light micrographs of hippocampus with cresyl violet staining in the control, KA, RE, and KA+RE groups in CA1 cell layer of hippocampus (400 $ \mathrm{\times} $). Black and white arrows indicate intact and degenerating neurons, respectively.

Fig.7.  Fluorescent micrographs of hippocampus with Fluoro-Jade B (FJB) from control, KA, RE, and KA+RE groups in CA1 cell layer of hippocampus (x400). Black and white arrows indicate the intact and fluorescent positive neurons, and white arrows indicate the neurons, respectively. Numerous FJBstained positive neurons in the CA1 cell layer are seen in KA group. Rare FJB-stained fluorescent cells are detected in CA1 cell layer by pretreatment of RE.

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