1 2588-4042 Iranian association of Physiology and Pharmacology 777 Respiratory system The cholinergic nervous system and lung diseases: dual inflammatory and anti-inflammatory effects Fatemikia Hossein b Ketabchi Farzaneh c b Dep of Physiology, School of Medicine, Shiraz University of Medical Sciences, Shiraz, Iran c Dep of Physiology, School of Medicine, Shiraz University of Medical Sciences, Shiraz, Iran 1 12 2023 7 1 18 28 10 2025 31 12 2023 Pulmonary diseases are among the most common diseases that are associated with the disability of patients and, in severe cases, with high mortality. In recent years, the role of the autonomic nervous system in aggravating or modulating the inflammatory reactions of various body organs, including the lungs, has been discussed. Some reports indicate the anti-inflammatory role of the parasympathetic nervous system, while other studies have suggested the inflammatory role of this system. Also, some studies highlight the role of the spleen in reducing inflammation, while others have considered the role of the spleen in exacerbating inflammation and tissue damage. Meanwhile, the role of alpha-7 nicotinic receptor in spleen and lung is controversial. The contradictory results of these studies can be attributed to the differences in the timing of stimulation and inhibition of the parasympathetic nervous system, the severity of the lung injury, and the type of pathogenic and non-pathogenic factors in different animal models of lung injury. In this review, we have discussed the role of the parasympathetic nervous system, spleen and a7 nicotinic acetylcholine receptor in the pathogenic and non-pathogenic lung diseases and the underlined mechanisms. According to the data of this article, more studies are needed to clarify the inflammatory or anti-inflammatory role of the cholinergic system in various inflammatory lung diseases.  
778 Neuroscience Effects of saffron hydro-alcoholic extract on the hippocampal oligodendrocyte transcription factor 2 and glial fibrillary acidic protein distribution in female demyelinated rats Javid Zahra d Taherianfard Mahnaz e Sajedianfard Javad f Moradi Hamid Reza g d Department of Basic Sciences, School of Veterinary Medicine, Shiraz University, Shiraz, Iran e Department of Basic Sciences, School of Veterinary Medicine, Shiraz University, Shiraz, Iran f Department of Basic Sciences, School of Veterinary Medicine, Shiraz University, Shiraz, Iran g Department of Basic Sciences, School of Veterinary Medicine, Shiraz University, Shiraz, Iran 1 12 2023 7 19 31 28 10 2025 31 12 2023 Background and aim: Demyelinating diseases are autoimmune disorders of the nervous system. The present study aims to effect of the hydro-alcoholic extract of saffron on blood brain-barrier destruction, Glial fibrillary acidic protein, and Oligodendrocyte transcription factor 2 distribution in astrocyte and oligodendroglia cells and the number of mentioned cells.  Methods: Forty female Sprague Dawley adult rats in four groups, Sham, treatment control, saffron treatment, and infliximab treatment were used. Demyelination was induced by 3 µl of 0.01% ethidium bromide injection in the dentate gyrus. Immunohistochemistry was used, for the distribution of Glial fibrillary acidic protein and Oligodendrocyte transcription factor 2 determination. The Image Analyzer software have evaluated the distribution of Glial fibrillary acidic protein and Oligodendrocyte transcription factor 2. Demyelination was determined by Luxol fast blue staining and image-j software. The ELISA method and Evans blue extravasation measurement have investigated the extent of blood brain-barrier disruption. Results: Blood brain-barrier disruption was meaningfully (p < 0.05) more in the treatment control compared to the other groups. The distribution of Glial fibrillary acidic protein and Oligodendrocyte transcription factor 2 and the number of both cells were considerably (p < 0.001) lower in the treatment control relative to the other groups. Glial fibrillary acidic protein and Oligodendrocyte transcription factor 2 remarkably (p < 0.05) were lesser in the treatment control compared to the other groups.  Conclusion: The hydro-alcoholic extract of saffron ameliorates blood brain-barrier destruction, improved Glial fibrillary acidic protein, and Oligodendrocyte transcription factor 2 distribution and number of astrocyte and oligodendroglia cells.   779 Neuroscience The role of central opioid and CB1 cannabinoid receptors in the antinociceptive effect of curcumin on the formalin-induced muscle pain in rats Naqshbandi Nabat h Tamaddonfard Esmaeal i Erfanparast Amir j Soltanalinejad-Taghiabad Farhad k h Department of Basic Sciences, Faculty of Veterinary Medicine, Urmia University, Urmia, Iran i Department of Basic Sciences, Faculty of Veterinary Medicine, Urmia University, Urmia, Iran j Department of Basic Sciences, Faculty of Veterinary Medicine, Urmia University, Urmia, Iran k Department of Basic Sciences, Faculty of Veterinary Medicine, Urmia University, Urmia, Iran 1 12 2023 7 32 45 28 10 2025 31 12 2023 Background and aim: Curcumin, the biologically active ingredient of turmeric (Curcuma longa), affects pain processing at different levels. Local mechanisms of action of curcumin are somewhat known, but the central effects at the spinal and supraspinal levels have been less investigated. In this study, the effect of oral administration of curcumin on the formalin-induced muscle pain was investigated. In addition, naloxone (a non-selective opioid receptor antagonist) and AM251 (a cannabinoid 1 receptor antagonist) were used to clarify the possible central mechanism of action of curcumin.  Methods: For intracerebroventricular (ICV) injections, one stainless steel guide cannula was implanted into in the left lateral ventricle of the brain in anaesthetized rats. Muscle pain was induced by intramuscular (IM) injection of formalin (2.5%, 50 µl) in the right gastrocnemius muscle, and the number of paw flinches and the time spent licking and biting of the injected paw were recorded at 5-min blocks for 1 h. Results: A marked biphasic pain response was observed due to formalin injection. Oral administration of curcumin at doses of 25 and 100 mg/kg significantly (p < 0.01 and p < 0.001, respectively) decreased both phases of muscle pain. Pain intensity was not influenced by alone ICV injection of naloxone and AM251 (p > 0.05), whereas the suppressive effect of 100 mg/kg curcumin was inhibited by ICV administration of 2 µg naloxone and 2 µg AM251 (p < 0.001). All the above-mentioned treatments did not alter locomotor activity. Conclusion: These results showed that curcumin produced an antinociceptive effect in muscle pain. Central opioid and cannabinoid receptors might be involved in the antinociception induced by curcumin. ‎   780 Endocrine Alterations of miR-148a, miR-223, and some blood lipoproteins following high-intensity functional training in overweight or obese middle-aged women shakib Amir l Zolfi Hamid Reza m Pashei Zhaleh n Moradi Leila o l 1. Faculty of Physical Education and Sport Sciences, University of Tabriz, Tabriz, Iran m 2. Department of Physical Education and Sport Science, Technical and Vocational University (TVU), Tehran, Iran n 1. Faculty of Physical Education and Sport Sciences, University of Tabriz, Tabriz, Iran o 1. Faculty of Physical Education and Sport Sciences, University of Tabriz, Tabriz, Iran 1 12 2023 7 45 56 28 10 2025 31 12 2023 Background and aim: MicroRNAs (miRNA) are single-stranded non-coding RNAs containing 19-22 nucleotides. Research investigating the molecular processes underlying obesity-induced dyslipidemia has revealed that miRNA plays a conflicting role in this scenario. As there is a need for more research in this area, this study aimed to investigate how high-intensity functional training (HIFT) affects miR-148a, miR-223, and blood lipoproteins in overweight or obese middle-aged women..  Methods: Twenty middle-aged women between 35-40 years old and overweight or obese (BMI=25-30 kg/m2) were randomly assigned to two groups based on their fat percentage and aerobic capacity (V̇O2max). The first group underwent HIFT exercises, while the second group served as a control. Blood samples were taken from the participants 48 hours before and after eight weeks of HIFT exercises to analyze miR-148a, miR-223, and blood lipid profiles. The data was then analyzed using analysis of covariance (ANCOVA), paired t-test, and Bonferroni post hoc tests. Results: After eight weeks of the HIFT training program, the training group showed a significant decrease miR-148a (p = 0.001), miR-223 (p = 0.006), LDL-C (p < 0.001), triglyceride (p = 0.002), and cholesterol (p < 0.001) compared to the control group. Additionally, the concentration of HDL-C (p = 0.001) increased in the training group compared to the control group. Conclusion: It seems that HIFT training can improve blood lipid profiles by affecting some obesity-related miRNAs, especially miR-148a miR-223, and improve health thereby preventing diseases caused by dyslipidemia such as type 2 diabetes, and cardiovascular disease. ‎   781 Endocrine The effect of Rosmarinic acid on occurrence of morphine withdrawal syndrome in mice Molladizavandi Mahla p Saberi Mehdi Poorheidari Gholamreza Mashhadi Akbar Boojar Mahdi p Student Research Committee, Baqiyatallah University of Medical Sciences, Tehran, Iran Department of Pharmacology and Toxicology, Faculty of Pharmacy, Baqiyatallah University of Medical Sciences, Tehran, Iran Department of Pharmacology and Toxicology, Faculty of Pharmacy, Baqiyatallah University of Medical Sciences, Tehran, Iran Department of Pharmacology and Toxicology, Faculty of Pharmacy, Baqiyatallah University of Medical Sciences, Tehran, Iran 1 12 2023 7 57 65 28 10 2025 31 12 2023 Background and aim: Evidence indicates the beneficial effects of Lemon balm polyphenols in controlling morphine withdrawal syndrome. The effects of Rosmarinic acid, as one of the most active biological components of this plant, have not been studied in the field of opioid withdrawal syndrome, and therefore, this study was conducted with the aim of investigating the effect of Rosmarinic acid on this syndrome in an animal model.  Methods: To induce dependence, morphine was injected intraperitoneally three times a day (with doses of 50, 50, and 75 mg/kg respectively) for 3 days. On the fourth day after morphine, naloxone was administered to induce the withdrawal syndrome. Different doses of Rosmarinic acid (5, 10, and 20 mg/kg) as active treatments, clonidine, and saline (negative control) were injected in different and separate groups (n = 8), 30 minutes before naloxone. The levels of anxiety, irritability, and pain tolerance threshold were evaluated by elevated plus maze (EPM), open field, and hot plate tests after naloxone injection. Results: Clonidine and Rosmarinic acid with a dose of 20 could significantly increase the presence of mice in the open arm of the EPM (p < 0.001). The excitability and motor activity in mice with withdrawal syndrome in the groups receiving 10 and 20 Rosmarinic acid and clonidine significantly decreased and the pain threshold increased (p < 0.05), and there was no significant difference between the Rosmarinic acid 20 and clonidine groups. Conclusion: These results clearly show that moderate to high doses of Rosmarinic acid (>10 mg/kg) are as effective as clonidine in suppressing the occurrence of morphine withdrawal syndrome and can be considered as a candidate in future clinical trials. ‎   782 Endocrine The effect of intrahippocampal injection of orexin A and orexin receptor antagonists 1 and 2 on passive avoidance memory, motor coordination and balance Shokoohi-Yekta Hamideh Khazali Homayoun Faculty of Life Sciences and Biotechnology, Shahid Beheshti University, Tehran, Iran Faculty of Life Sciences and Biotechnology, Shahid Beheshti University, Tehran, Iran 1 12 2023 7 66 74 28 10 2025 31 12 2023 Background and aim: The aim of the research was to block orexin receptors in the hippocampus and investigate the changes in passive avoidance memory, motor coordination and balance in the absence of orexin effects. The effect of intra-hippocampal injection of the orexin receptor type 2 antagonist JNJ-10397049 on passive avoidance memory, motor coordination and balance was not investigated in previous studies, which we addressed in this research. Methods: Twenty adult male Wistar rats weighing 220-250 grams were randomly divided into four groups. Animals in each group respectively received saline, Orexin A, Orexin A and Orexin receptor type 1 antagonist, or Orexin A and Orexin receptor type 2 antagonist in their hippocampus. Subsequently, passive avoidance memory was evaluated using a shuttle box, while motor coordination and balance were assessed utilizing rotarod devices. Results: The average delay in entering the dark area and the total time that the animals spent in the light area of the shuttle box increased in the orexin-receiving group compared to the control group. However, these values significantly decreased in the groups that received both orexin and orexin receptor antagonists, in comparison to the group that received orexin alone (p < 0.01). Furthermore, the duration of motor coordination and balance impairment 30 minutes after injection increased in the orexin-treated group compared to the control group, but it showed a significant decrease (p < 0.01) in the groups that received both orexin and orexin receptor antagonists, when compared to the group that received orexin alone. Conclusion: It seems that orexin in the hippocampus improves the retrieval of passive avoidance memory, motor coordination and balance. On the other hand, Orexin receptor antagonists disrupt these two processes. ‎   783 Endocrine Cadmium and Mercury effects on the stress oxidative and AHR-CYP1A1 pathway in liver of mice Soltan-Oghli Mehdi Mohammadi-Bardbori Afshin Hassani Shokoufeh Shadboorestan Amir Ghafarian-Bahreman Ali Keshavarzi Majid Omidi Mahmoud Student Research Committee, Faculty of Pharmacy, Shiraz University of Medical Sciences, Shiraz, Iran Department of Pharmacology and Toxicology, Faculty of Pharmacy, Shiraz University of Medical Sciences, Shiraz, Iran Toxicology and Diseases Group (TDG), Pharmaceutical Sciences Research Center (PSRC), the Institute of Pharmaceutical Sciences (TIPS), Tehran, Iran Department of Toxicology, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, Iran Occupational Environment Research Center, Rafsanjan University of Medical Sciences, Rafsanjan, Iran Department of Pharmacodynamics and Toxicology, Faculty of Pharmacy, Mashhad University of Medical Sciences, Mashhad, Iran Department of Pharmacology and Toxicology, Faculty of Pharmacy, Hormozgan University of Medical Sciences, Hormozgan, Iran 1 12 2023 7 75 83 28 10 2025 31 12 2023 Background and aim: Aryl Hydrocarbon Receptor (AHR) is an environmental sensor and sensitive to oxidative change. However, little is known about the effects of stress, especially oxidative stress, on aryl hydrocarbon receptor signaling. Therefore, this study aimed to investigate the effects of oxidizing compounds on the function of aryl hydrocarbon receptor in mice. Methods: For this purpose, in mice treated with Formylindolo[3,2-b] carbazole (FICZ), buthionine sulfoximine (BSO), cadmium (Cd), and mercury (Hg), the effects of oxidants on the function of aryl hydrocarbon receptors were studied for 3 and 24 hours. Results: For the first time, the data of this research unveil a clear relationship between Cytochrome P450 Family 1 Subfamily A Member 1 (CYP1A1) activity and cellular oxidative status. The in vivo findings indicated that the duration of activation/inhibition of aryl hydrocarbon receptors was directly related to the increase/decrease of the reduced glutathione/oxidized glutathione (GSH/GSSG) ratio. Conclusion: This study showed that many environmental pollutants such as heavy metals and oxidants might affect the normal function of AHR target enzymes like CYP1A1 by altering intracellular oxidative potential.   784 Cellular and Molecular The effect of Echium amoenum hydro-methanolic extract on MCF-7 and MDA-MB468 breast cancer cell lines and increasing the cytotoxicity of doxorubicin Fazeli Mehdi Department of Basic Sciences, School of Veterinary Medicine, Shiraz University, Shiraz, Iran 1 12 2023 7 84 94 28 10 2025 31 12 2023 Background and aim: Chemotherapy is one of the most common and important treatments in the fight against breast cancer, but drug resistance to these drugs reduces the success of the treatment. Today, extensive research is focused on the role of herbal compounds in increasing the effect of chemotherapeutic drugs. This study was conducted to investigate the effect of hydro-methanolic extract of Echium amoenum on two breast cancer cell lines MCF-7 and MDA-MB468 and its role in the cytotoxic activity of doxorubicin. Methods: The effect of Echium amoenum extract and doxorubicin alone or in combination were examined at two incubation times of 24 and 48 hours. Cells were seeded at a density of 104 cells per well in 96-well plates, and after treatment with different concentrations of extract and drug, their viability was measured by MTT assay. Results: Echium amoenum extract showed a high cytotoxic effect on breast cancer cells, while it had less toxicity on normal cells. The extract at the highest tested concentration reduced the percentage of viable cells in MCF-7 cells and in MDA-MB468 cells to 10.59 ± 0.72 and 12.22 ± 0.78%, respectively. The inhibitory effect of doxorubicin was 19.31 ± 0.92 and 18.13% ± 0.99 for MCF-7 and MDA-MB468 cells, respectively. IC50 of the extract in 24 and 48 hours incubation was calculated, for MCF-7 cells, 2.41 and 1.78 mg/ml, respectively, and for MDA-MB468 cells, 2.41 and 1.78 mg/ml, respectively.. The extract also showed a significant effect in increasing the cytotoxicity of doxorubicin. The maximum effect was seen with a concentration of 5 mg/ml of extract on a concentration of 1 μg/ml of doxorubicin in 48 hours incubation. The percentage of survival in MCF-7 and MDA-MB468 cells was decreased from 46.38 ± 1.80 to 24.23 ± 1.46 and from 55.30 ± 1.95 to 35.99 ± 1.40, respectively. Conclusion: Echium amoenum extract effectively killed the breast cancer cells in vitro and increased the cytotoxicity of doxorubicin. Further evaluation of the anti-tumor effects of the extract can be done in vivo.   785 Neuroscience Examining two practical methods for modeling relapse in preclinical studies Aghajani Niloofar Hashemizadeh Shiva Azizi Hossein Semnanian Saeed 1Department of Physiology, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, Iran 2School of Cognitive Sciences, Institute for Research in Fundamental Sciences, IPM, Tehran, Iran 1Department of Physiology, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, Iran 1Department of Physiology, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, Iran 1 12 2023 7 95 108 28 10 2025 31 12 2023 Drug addiction is a recurring chronic disease characterized by repeated and compulsive drug use and seeking behavior, despite awareness of its negative consequences. Relapse, which refers to a person's return to old patterns of behavior and drug use after a period of abstinence, is a significant concern in addiction treatment. One of the main concerns in addiction treatment is the high rate of relapse, even during a short period after withdrawal. It is common for individuals to experience relapse shortly after completing treatment. Exposure to the drug itself, drug-related cues and coping with stress are three factors induce relapse. To investigate and address the issue of relapse, significant efforts have been made to develop extinction-reinstatement models in laboratory animals. Modeling a human psychological disorder such as addiction in animals is not possible due to its complexity, laboratory animals can be used to simulate human psychological issues objectively and behaviorally, with measurable parameters. Over the past decades, animal models of relapse have been developed to investigate and address this neurobiological phenomenon. These models primarily focus on seeking behavior and drug taking, using methods such as spontaneous administration of substances and conditional place preference. This review article provides description of these two main models in relation to the "reinstatement" phenomenon.   786 Neuroscience Investigating the central role of the dopaminergic system in the physiological functions of the body: what we know so far Mahdavi Kimia Zendehdel Morteza Department of Basic Sciences, Faculty of Veterinary Medicine, University of Tehran, Tehran, Iran Department of Basic Sciences, Faculty of Veterinary Medicine, University of Tehran, Tehran, Iran 1 12 2023 7 109 122 28 10 2025 31 12 2023 The dopaminergic system plays an important role in modulating some neural functions such as motor control, motivation, pain, cognition, maternal behavior, and reward. Dopaminergic signaling pathways are very important for maintaining physiological processes, and an unbalanced activity may lead to disorders related to neurological diseases. A correct understanding of the neurobiology and underlying molecular mechanisms of these disorders leads to the development of new treatments that improve the quality of life of patients all over. The purpose of the present study is to review the research conducted on the central role of the dopaminergic system and the dimensions of its influence on various physiological functions. In this review, articles related to the topic in the years 1970 to 2023 were identified using a targeted search of related keywords in reliable databases such as Civilica, Web of Science, Scopus, Science Direct, PubMed, Springer, Google Scholar, and Elsevier, and in order to obtain more studies from the list The sources of these articles were used. Based on the findings of these studies, the appetite-reducing effects of dopamine have been reported in birds and mammals. On the other hand, the role of this neural mediator in pain perception and memory processing was determined. In connection with the endocrine glands, the interaction of dopamine with prolactin, sex steroids, growth hormone, and thyroid hormones has been proven. Also, numerous studies have shown the role of dopamine in motor control and the occurrence of maternal behavior. Finally, based on the review of past research, the occurrence of neurological diseases such as schizophrenia, Parkinson, Huntington, attention deficit/hyperactivity disorder (ADHA), and addiction can be related to abnormalities in the dopaminergic system. Despite conducting numerous studies on the role of the dopaminergic system in various biological activities, it is suggested to carry out more studies in order to identify new dimensions of the effect of this system on different body functions and to provide new treatment paths. ‎   657 Others... Nanobodies and methods of increasing their half-lives Khodabakhsh Farnaz Salimian Morteza Ahangari Cohan Reza 1Department of Genetics and Advanced Medical Technology, Medical Biotechnology Research Center, Faculty of Medicine, AJA University of Medical Sciences, Tehran, Iran 2Department of Clinical Laboratory Sciences, Faculty of Allied Medical Sciences, Kashan University of Medical Sciences, Kashan, Iran Department of Nanobiotechnology, New Technologies Research Group, Pasteur Institute of Iran, Tehran, Iran 1 12 2023 7 123 138 15 11 2023 31 12 2023 The development of antibody fragments is growing due to problems associated with commercially available antibodies including their large size and expensive and time-consuming production processes. Nanobodies are fragmented antibodies that are found naturally in the serum of the Camelidae family. Nanobodies, as the smallest functional part of antibodies, have many applications in the diagnosis and treatment of various diseases, especially in tumors. However, their small size causes them to be quickly removed from the blood circulation after administration, which restricted their use in the clinics. To solve this issue, various methods have been proposed to increase their plasma half-life, generally using two mechanisms: (1) the endosomal recycling of neonatal Fc receptor and (2) the increase of the hydrodynamic volume. The first mechanism naturally occurs in the body and leads to an increase in the plasma half-life of immunoglobulins and albumin. The chemical conjugation of nanobodies to the synthetic polymers, genetic fusion to polymer-mimetics peptides, and dimerization of the nanobodies increase the plasma half-life of nanobodies via the second mechanism. In this review, first, we will introduce the nanobodies, and in the following, we will discuss various methods that have been developed to increase their plasma half-lives.   662 Neuroscience Paradoxical effects of menthol on passive avoidance and spatial memory in the rat Armand Raham Vatanparast Jafar 1Department of Biology, School of Science, Shiraz University, Shiraz, Iran 1Department of Biology, School of Science, Shiraz University, Shiraz, Iran 1 12 2023 7 139 149 07 12 2023 31 12 2023 Background and aim: Menthol is well-known for its peripheral effects, e.g. cold receptor stimulation, pain modulation, and anti-inflammatory properties. However, its impacts on the CNS remain less understood. Here, we studied the acute and chronic effects of menthol, on passive avoidance learning-retention, and spatial memory, respectively. Methods: An appropriate dose of menthol that did not acutely affect motor activity was determined. Rats were then administered either a single dose of menthol or the solvent via intraperitoneal injection before the training or test session. In one group, bicuculline (1 mg/kg) was coadministered with menthol to rats prior to the training session. To determine the chronic effect of menthol on spatial memory, rats received daily doses of menthol or the solvent for 12 days before each session of the spatial memory test. Results: A dose of 40 mg/kg of menthol did not significantly alter motor activity in the open field test, but higher doses caused significant decrease of locomotion. Administration of menthol (40 mg/kg) before the training session led to reduced learning and memory retention. This effect was found to be inhibited by bicuculline. However, menthol administration before the test session had no effect on fear memory retention. Chronic administration of menthol during the spatial memory test period improved reference and working spatial memory. Conclusion: These findings suggest that menthol impairs fear memory learning and retention by activating GABA receptors. Menthol may also improve spatial memory through its reported anxiolytic action. Further studies are necessary to confirm this potential mechanism. 663 Neuroscience Exposure to morphine during embryonic period impairs learning in attention task in rats Alaee Elham Azizi Hossein Department of Physiology, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, Iran Department of Physiology, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, Iran 1 12 2023 7 150 158 07 12 2023 31 12 2023 Morphine passes through the placenta and selectively accumulates in the nervous tissues of the fetus. In the present study, the effect of exposure to morphine during embryonic period on cognitive functions such as attention and impulsive behaviors in male offspring has been investigated. Methods: Female rats received morphine subcutaneously twice a day in doses of 5 and 10 mg/kg during pregnancy (days 11 to 18). In the behavioral study, the 5-CSRTT (5-choice serial reaction time task) was used. Results: In the training phase of the behavioral test, the prenatal morphine group needed more days to learn the test than the saline group. In the baseline condition, the animals exposed to morphine in the embryonic period showed more impulsive and obsessive responses compared to the saline group. Conclusion: Behavioral results show a direct effect of exposure to morphine during embryonic period on delayed learning and impulsive behaviors in adulthood. 665 Neuroscience Investigating the mechanism of protective effects of fingolimod administration following hypoxia-induced neonatal seizures in adult male and female rats: the possible role of brain-derived neurotrophic factor and nitric oxide Bavarsad Kowsar Yazdanfar Neda Farbood Yaghoob Sarkaki Alireza Ghafouri Samireh 1. Department of Physiology, Faculty of Medicine, Ahvaz Jundishapur University of Medical Sciences, Ahvaz, Iran 2. Persian Gulf Physiology Research Center, Medical Basic Sciences Research Institute, Ahvaz Jundishapur University of Medical Sciences, Ahvaz, Iran 3. Neuroscience Research Center, Tabriz University of Medical Sciences, Tabriz, Iran 1. Department of Physiology, Faculty of Medicine, Ahvaz Jundishapur University of Medical Sciences, Ahvaz, Iran 2. Persian Gulf Physiology Research Center, Medical Basic Sciences Research Institute, Ahvaz Jundishapur University of Medical Sciences, Ahvaz, Iran 1. Department of Physiology, Faculty of Medicine, Ahvaz Jundishapur University of Medical Sciences, Ahvaz, Iran 2. Persian Gulf Physiology Research Center, Medical Basic Sciences Research Institute, Ahvaz Jundishapur University of Medical Sciences, Ahvaz, Iran 1. Department of Physiology, Faculty of Medicine, Ahvaz Jundishapur University of Medical Sciences, Ahvaz, Iran 2. Persian Gulf Physiology Research Center, Medical Basic Sciences Research Institute, Ahvaz Jundishapur University of Medical Sciences, Ahvaz, Iran 1 12 2023 7 159 168 21 12 2023 31 12 2023 Background and aims: Hypoxia-induced neonatal seizures activate neuroinflammatory and oxidative stress mechanisms and lead to brain damage. Therefore, considering the high sensitivity of the immature brain to anaerobic metabolism during hypoxia, the purpose of this study is to investigate the mechanism of the protective effects of fingolimod in modulation of complications caused by neonatal seizures induced by hypoxia in puberty. Methods: In this study, 40 male and female wistar rats were divided into 4 groups: 1. control-saline, 2. control-fingolimod, 3. hypoxia-saline and 4. hypoxia-fingolimod. Neonatal seizures were induced 10 days after birth by placing the neonates in hypoxia chamber for 15 minutes. In different experimental groups, fingolimod or saline were injected from the 10th to the 21st day after birth. Then, 60 days after birth, hippocampus samples were extracted and used for biochemical evaluations. Results: The results of this study showed that fingolimod significantly prevents the increased concentration of brain-derived neurotrophic factor and nitric oxide in the hippocampus of both male and female hypoxia groups (p < 0.05). Although this decrement was not significant in hypoxia-fingolimod female group. Conclusion: Administration of fingolimod can modulate complications caused by neonatal seizures, possibly through decreasing the expression of brain-derived nerve growth factor and the compensatory effect of nitric oxide during puberty.   667 Neuroscience The effect of long-term deprivation of Whisker on the lateralization of cerebral hemispheres and the discrimination of surfaces in male rats Haghpanah Tahereh Meftahi Gholamhosein Shojaeian Melika Afarinesh Mohammad Reza Anatomical Sciences Department, School of Medicine, Kerman University of Medical Sciences, Kerman, Iran Neuroscience Research Center, Baqiyatallah University of Medical Sciences, Kerman, Iran Neuroscience Research Center, Institute of Neuropharmachology, Kerman University of Medical Sciences, Kerman, Iran Neuroscience Research Center, Institute of Neuropharmachology, Kerman University of Medical Sciences, Kerman, Iran 1 12 2023 7 169 177 27 12 2023 31 12 2023 Background and aims: The whisker movements in rodents project sensory information to the barrel cortex, allowing the animal to perceive its environment. Deprivation of whiskers may affect the animal's instinctive behavior towards the left or right side of the body. This study examined the impact of two months of whisker deprivation on brain hemisphere dominance and tissue level differentiation in rats. Methods: Firstly, male Wistar rats were unilaterally deprived of whiskers (left or right) and bilaterally from birth to 60 days old. Then, the animals' preference was then studied using the tail suspension test at one month old, and the difference in roughness of two-month-old animals was evaluated using the tactile discrimination test. Results: Animals with unilateral whisker deprivation (right or left) showed a more tendency to turn left compared to the control and bilateral whisker-deprived groups. Additionally, the ability to distinguish roughness in both unilateral and bilateral whisker-deprived groups was impaired compared to the control group (p < 0.05). Conclusion: Deprivation of whiskers reduces touch discriminations and can affect rats' preference for lateral superiority instinctive behavior.   668 Others... Dopamine facilitates toxoplasma tachyzoite replication by stimulating D1-like dopamine receptors Yasami-Khiabani Setayesh Mostafavi Maryam Babaie Jalal Golkar Majid Department of Parasitology, Pasteur Institute of Iran, Tehran, Iran Department of Parasitology, Pasteur Institute of Iran, Tehran, Iran Department of Parasitology, Pasteur Institute of Iran, Tehran, Iran Department of Parasitology, Pasteur Institute of Iran, Tehran, Iran 1 12 2023 7 178 186 06 01 2024 31 12 2023 Background and Aim: Toxoplasma gondii is an obligate intracellular parasite. T. gondii leads to increased production of dopamine in the host's brain. It is possible that dopamine plays a role in the proliferation of T. gondii. In this study, the effect of dopamine on the proliferation of Toxoplasma tachyzoites in HFF-1 cells was investigated. Methods: HFF-1 cells were cultured and treated with 500 nM dopamine. 4 × 105 tachyzoites were added to the cell wells. To investigate the role of receptors, first the confluensed cells were treated with dopamine. Dopamine D1 and D2 receptor antagonists (SCH-23390 and sulpiride, respectively), alone and in combination, were added to the culture medium at a concentration of 10 nM. The treated cells were infected with Tachyzoite. The amounts of tachyzoites were evaluated by microscopic counting and by RT-qPCR method. Results: Dopamine significantly (p < 0.05) increases proliferation of tachyzoites in HFF-1 cells. SCH-23390 significantly neutralized the effect of dopamine in treated cells (p < 0.05), while sulpiride could not avert the effect of dopamine. Moreover, combination of two antagonists, SCH-23390 and sulpiride, significantly reduced the proliferation of tachyzoites in HFF-1 cells (p < 0.05). Conclusion: Our study showed that dopamine stimulates proliferation of tachyzoites in HFF-1 cell line. In addition, dopamine exerts its effect on parasite proliferation through D1 receptors. These results can help to design and develop effective drugs against Toxoplasma.   671 Neuroscience Permanent effects of maternal deprivation stress and adolescent morphine exposure on anxiety-like behaviors and working memory in male and female rats Khani Fatemeh Mahmoudi Ershad Zeinivand Motahareh Nedaei Seyed Ershad Sleep Disorders Research Center, Kermanshah University of Medical Sciences, Kermanshah, Iran Department of Physiology, Kermanshah University of Medical Sciences, Kermanshah, Iran Department of Physiology, Kermanshah University of Medical Sciences, Kermanshah, Iran Department of Physiology, Kermanshah University of Medical Sciences, Kermanshah, Iran 1 12 2023 7 187 196 15 01 2024 31 12 2023 Background and Aim: Experience of separation from mother can cause lasting changes in brain structure. Meanwhile, exposure to opioids in adolescence can affect cognitive abilities. In this study, effects of maternal deprivation stress and exposure to morphine in adolescence on anxiety-like behaviors and working memory were evaluated in adult rats. Methods: Male and female Wistar rats including control, maternal deprivation stress (Str), receiving morphine (Morphine) in adolescent, and Morphine with Stress (Mor + Str) group were selected. In the Str group, the offspring was separated from their mothers 3 hours/day for 14 days. Morphine group received increasing doses of morphine from 2.5 to 25 mg/kg for 10 days (31-40 postnatal day). Open field test, elevated plus maze and Y maze were used to investigate locomotor activity, anxiety-like behaviors and working memory of animals, respectively. Results: There was no significant difference in locomotor activity between morphine and control groups. Morphine group in both sexes spent more time in center of the open field compared to control group, and unlike the Mor + Str group, had stable anti-anxiety behavioral effects until adulthood. Working memory was impaired in both Morphine and Mor + Str groups in both sexes compared to control group. Conclusion: Maternal deprivation and substance abuse have lasting effects on occurrence of anxiety-like behaviors during the developmental period of adolescence. Maturation of the nervous system can be affected by these adverse environmental factors and underwent permanent behavioral changes and cognitive disorders.   674 Others... Preparation of Polyclonal Antibody against B-cell activating factor Mardani-Jouneghani Rasoul Irani Shiva Habibi-Anbouhi Mahdi Behdani Mahdi Department of Biology, Science and Research Branch, Islamic Azad University, Tehran, Iran Department of Biology, Science and Research Branch, Islamic Azad University, Tehran, Iran National Cell Bank of Iran, Pasteur Institute of Iran, Tehran, Iran Biotechnology Research Centre, Venom and Biotherapeutics Molecules Laboratory, Pasteur Institute of Iran, Tehran, Iran 1 12 2023 7 197 205 04 02 2024 31 12 2023 Background and Aim: B-cell activating factor (BAFF) belongs to the tumor necrosis factor (TNF) superfamily and plays a critical role in B-cell survival and differentiation. Overexpression of BAFF has been linked to autoimmune diseases and B-cell malignancies. The biologically active segment of this protein is a soluble domain, making it a promising target for antibody-based therapies. This study aimed to develop a polyclonal camel antibody capable of detecting BAFF on cell surfaces. Methods: An expression vector, pET22b, containing the extracellular domain of the BAFF protein (NM_001145645.2), was constructed and introduced into Escherichia coli BL21 (DE3) using the calcium chloride heat shock method (CaCl2). Subsequently, the recombinant protein was induced using 1mM IPTG and protein purification was carried out with Ni2+–NTA resin. A 9-month-old female Camelus dromedaries was immunized with purified recombinant BAFF protein (rBAFF) mixed with Freund's adjuvant. Following immunization, the isolated polyclonal antibody was assessed using ELISA, western blotting, and flow cytometry to detect rBAFF protein. Results: The study confirmed the successful preparation of recombinant BAFF protein and the effectiveness of camel immunization and purification processes. The isolated polyclonal antibody demonstrated the ability to identify recombinant BAFF proteins in ELISA and western blot tests. Flow cytometry demonstrated its capacity to detect BAFF protein on cell surfaces. Conclusion: Given the significance of diagnosing and developing novel adjuvant treatment methods for autoimmune diseases and cancer, this study's findings support the potential use of polyclonal antibodies targeting the BAFF antigen as valuable tools for these purposes. 675 Neuroscience The effect of safranal on epileptiform activity induced by penicillin: Role of somatosensory cortex GABAA-benzodiazepine receptor complex Tamaddonfard Sina Erfanparast Amir Tamaddonfard Esmaeal Soltanalinejad-Taghiabad Farhad Department of Basic Sciences, Faculty of Veterinary Medicine, Urmia University, Urmia, Iran Department of Basic Sciences, Faculty of Veterinary Medicine, Urmia University, Urmia, Iran Department of Basic Sciences, Faculty of Veterinary Medicine, Urmia University, Urmia, Iran Department of Basic Sciences, Faculty of Veterinary Medicine, Urmia University, Urmia, Iran 1 12 2023 7 206 218 04 02 2024 31 12 2023 Background and Aim: Safranal is one of the bioactive compounds of saffron and has analgesic, anti-inflammatory, anti-diabetic and anti-cancer properties. In a variety of pathological states of the nervous system, safranal has shown a powerful neuroprotective effect. In this study, the effect of safranal on penicillin-induced epileptic activity was investigated. GABA-benzodiazepine receptor complex antagonist (flumazenil) was used to follow the possible mechanism of action. Methods: Epileptiform activity was induced by intra-somatosensory cortex microinjection of penicillin (300 IU, 1.5 μL) in urethane-anesthetized rats. Electrocorticographic recordings were used to record the latency time to first spike appearance and to analyze the frequency and amplitude of spike waves. Safranal was intraperitoneally injected and flumazenil and diazepam (as a positive control group) were intracerebroventricularly administered. Results: The first spike wave was appeared 3.74±0.27 min after penicillin microinjection. Penicillin also produced 31.91±0.99 spike wave per min with an amplitude of 1.22±0.03 mV. Intraperitoneal injection of safranal (1 and 4 mg/kg) and intracerebroventricular administration of diazepam (5 µg per animal) reduced both the number and amplitude of spike waves. Intracerebroventricular injection of flumazenil (5 µg per animal) prevented diazepam (5 µg per animal)-, and safranal (4 mg/kg)-induced anti-epileptiform activity. Conclusion: The results indicated an anti-epileptiform activity for safranal. It was also found that the central GABAA-benzodiazepine receptor complex might be involved in the antiepileptic activity of safranal.   683 Exercise physiology Effect of 8 weeks high-intensity interval training on the stress of liver endoplasmic reticulum in obese rats with type 2 diabetes Bayat Hadis Gholami Mandana Rajabi Hamid Abed Natanzi Hossein Department of physical education and sport science, Faculty of Literature, Humanities and Social Sciences, Science and Research Branch, Islamic Azad University, Tehran, Iran Department of Professional Physical Education and Sport Science, Kharazmi University, Tehran, Iran Department of physical education and sport science, Faculty of Literature, Humanities and Social Sciences, Science and Research Branch, Islamic Azad University, Tehran, Iran Department of physical education and sport science, Faculty of Literature, Humanities and Social Sciences, Science and Research Branch, Islamic Azad University, Tehran, Iran 1 12 2023 7 219 228 10 03 2024 31 12 2023 Background and Aim: Recent research has shown that stress of endoplasmic reticulum (ER) in liver tissue plays a significant role in development of type 2 diabetes. The aim of this study was to investigate the effect of 8 weeks high-intensity interval training on two known markers of liver ER stress in rats with type 2 diabetes. Methods: Twenty four male Wistar rats (8 weeks age and 200-250 g weight) were used. The "healthy control" group (8 mice) had free access to standard food and drinking water for 13 weeks. Another 16 mice were fed a high-fat diet for 4 weeks. Then these 16 rats were made diabetic with streptozotocin and allocated in two groups: "diabetic control" (maintenance of a high-fat diet for 9 weeks without exercise) and "diabetic exercise" (maintenance of a high-fat diet, one week familiarization with treadmill and then 8 weeks high intensity interval training). Then, the expression levels of eukaryotic initiator factor 2-alpha (eIF-2-α) and tumor necrosis factor alpha (TNF-α) in the liver tissue of all rats were measured by western blot method. Results: Blood glucose level and expression of both eIF2-α and TNF-α were significantly higher in the "diabetic control" than the "healthy control" (P≤0.05). The "diabetic exercise" group showed significantly lower glucose level than the the "diabetic control" group (P≤0.05). However, neither eIF-2-α nor TNF-α showed significant change in "diabetic exercise" compared to the "diabetic control" group. Conclusion: Intense intermittent exercise with the parameters used in this research could not change two stress markers of liver ER in type 2 diabetic rats.   685 Neuroscience Cuprizone causes the destruction of neuron myelin in the hippocampus through an oxidative stress-dependent process, yet it does not impair short-term spatial memory Gharighnia Sanaz Omidi Ameneh Department of Anatomical Sciences, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, Iran Department of Anatomical Sciences, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, Iran 1 12 2023 7 229 239 16 03 2024 31 12 2023 Background and Aim: Multiple Sclerosis (MS) is a demyelinating disease of the central nervous system that affects patients with many sensory, motor, and cognitive symptoms. This study aims to evaluate memory function and histopathology and measure the oxidative stress level in the hippocampus of the MS model in mice. Methods: Mice were divided into two groups that received a normal diet (control) and one that received a diet containing 0.2% cuprizone (cuprizone) for six weeks. After measuring the animal's weight and behavioral memory test, the hippocampus was evaluated in terms of histopathology and biochemical markers to measure the level of oxidative stress parameters. Results: The weight gain of mice after 6 weeks of the cuprizone diet was significantly lower than the control group (p < 0.05). Although behavioral evaluations performed with the Y maze test did not have significant differences between the two groups, the hippocampus in the cuprizone group revealed obvious demyelination. Measuring the parameters related to the level of oxidative stress showed that the amount of lipid peroxidation in the cuprizone group increased significantly compared to the control group (p < 0.05). In addition, the reduced glutathione and catalase activity levels in the cuprizone group showed a significant decrease compared to the control group (p < 0.05). Conclusion: The induction of acute demyelination with cuprizone, probably through the creation of oxidative stress in the hippocampus, was able to lead to demyelination in the tissue of this region, while it had no significant effect on the memory performance of the animals.   684 Exercise physiology Effects of a period of moderate-intensity interval training (MIIT) with ginger supplementation on PGC-1α and ATPIF1 gene expression in gastrocnemius muscle of aged male rats Ramezani Vavdare Fatemeh Nazarali Parvaneh Shakibaee Abolfazl Kazemi Fahimeh 1Department of Exercise Physiology, Faculty of Sport Sciences, Alzahra University, Tehran, Iran 1Department of Exercise Physiology, Faculty of Sport Sciences, Alzahra University, Tehran, Iran 2Exercise Physiology Research Center, Lifestyle Research Institute, Baqiyatallah University of Medical Sciences, Tehran, Iran 1Department of Exercise Physiology, Faculty of Sport Sciences, Alzahra University, Tehran, Iran 1 12 2023 7 240 249 16 03 2024 31 12 2023 Background and Aim: Peroxisome proliferator-activated receptor gamma coactivator 1-alpha (PGC-1α) and ATPase inhibitory factor 1 (ATPIF1) play a role in mitochondrial biogenesis. The aim of the present study was to determine the effects of a period of moderate-intensity interval training (MIIT) with ginger supplementation on PGC-1α and ATPIF1 gene expression in gastrocnemius muscle of aged male rats. Methods: In an experimental research, 20 aged male Wistar rats (22-month-old with a weight of 500 ± 100 g) were randomly divided into four groups of 5 each: control, interval training, ginger, and interval training + ginger. Training protocol was conducted on a treadmill for 8 weeks, three times a week. Ginger extract was orally administered to rats at a dose of 100 mg/kg body weight per day for 8 weeks. Forty-eight hours after last training session, tissue removal was performed from gastrocnemius muscle to measure the expression of PGC-1α and ATPIF1 genes. Non-parametric Kruskal-Wallis test was used for inter-group comparisons, and non-parametric Mann-Whitney U test was used to compare two groups. Results: Only a significant difference was observed in PGC-1α gene expression between interval training + ginger and control groups (p < 0.05). Also, there was no significant difference in ATPIF1 gene expression between research groups (p > 0.05). Conclusion: Eight weeks of MIIT with ginger supplementation can lead to an increased PGC-1α gene expression in gastrocnemius muscle of aged male rats, but it did not cause a significant change in ATPIF1 gene expression   689 Neuroscience The impact of high-frequency stimulation of the hippocampal Schaffer collateral pathway on the consolidation of spatial memory in male laboratory rats Asgari Tara Asgari-Mobarakeh Karim Sayyah Mohamad Gholami Pourbadie Hamid Department of Physiology and Pharmacology, Pasteur Institute of Iran , Tehran, Iran Department of Psychology, Faculty of Education and Psychology, University of Isfahan, Isfahan, Iran Department of Physiology and Pharmacology, Pasteur Institute of Iran , Tehran, Iran Department of Physiology and Pharmacology, Pasteur Institute of Iran , Tehran, Iran 1 12 2023 7 250 259 08 04 2024 31 12 2023 Background and Aim: The acquisition and consolidation of new tasks into long-term memory involve the appearance of sharp-wave ripples in the temporal lobe during resting or sleep, which play a crucial role in memory consolidation. Additionally, sharp-wave ripples, high-frequency waves in the molecular layer of the hippocampus, are associated with synaptic plasticity and memory consolidation. This study aims to investigate the impact of high-frequency electrical stimulation of the hippocampal Schaffer collateral (SC) pathway on memory consolidation in behaving rats. Methods: Stereotaxic surgery was conducted to implant stimulating electrodes in the SC pathway and recording electrodes in the CA1 region of male Wistar rats. After confirming electrophysiological recording, electrodes were secured to the skull using dental cement. Two hours following each acquisition session rats underwent stimulation of the SC pathway with a frequency of 200 Hz to assess its effect on spatial memory formation. Spatial learning was then conducted using the Morris water maze over three consecutive days. Electrical stimulation occurred two hours post-learning on each day, and spatial memory was evaluated on the fourth day. Results: Stimulation of the SC pathway with high frequency resulted in long-term synaptic strengthening in CA1. However, behavioral testing revealed no significant difference in learning and spatial memory between the group receiving electrical stimulation and the control group. Conclusion: These findings suggest that high-frequency stimulation of the SC pathway does not enhance memory, indicating that synaptic strengthening in the CA1 region does not necessarily translate into memory improvement. 691 Others... Factors affectingresearch and development and the role of research centers with a focus on comparing Iranian and other world institutions Mousavi Nafiseh Sadaat Sardari Soroush Drug Design and Bioinformatics Unit, Medical Biotechnology Department, Biotechnology Research Center, Pasteur Institute of Iran, Tehran, Iran Drug Design and Bioinformatics Unit, Medical Biotechnology Department, Biotechnology Research Center, Pasteur Institute of Iran, Tehran, Iran 1 12 2023 7 260 280 14 04 2024 31 12 2023 Research institutions emerged at the beginning of the 20th century. These institutions did their work independently and far from public services and government supervision. Such as the Rockefeller Institute, the Carnegie Institution of Washington and the Institute for Advanced Studies that emerged in the United States. As of 2006, there were more than 14,000 research centers in the United States. The National Institutes of Health in the United States, the Max Planck Instituteof Germany, the International Rice Research Institute of the Philippines, the European Molecular Biology Laboratory of Germany, and the National Cancer Institute are some of the world-class research institutions. The history of research centers in Iran dates back to the early 20th century, when the first scientific associations were established in major cities such as Tehran, Isfahan, and Shiraz. One of the most prominent research centers in Iran is the Institute of Basic Sciences, which was established in Tehran in1368. Another important research center in Iran is the National Institute of Medical Research, which was established in Tehran in1378. Research centers have made a significant contribution to scientific knowledge, promoting innovation and dealing with global challenges, and they continue to play an important role in advancing scientific research and education. These centers contribute to innovation and address global challenges by conducting research, promoting scientific education and spreading knowledge among scientists. The role and duties of research centers can be very different depending on their focus and mission. Some research centers are dedicated to basic scientific research, while others focus on applied research with practical applications. Many research centers are affiliated with universities or other academic institutions, while others are independent organizations. The functions of research centers may include conducting research, disseminating knowledge through publications and conferences, providing education and training to researchers and students, collaborating with industry and other organizations, and participating in policy and decision-making processes. The specific goals and objectives of a research center depend on its focus and mission, as well as the needs and priorities of the community it serves. We intend to compare Iran's research centers in terms of financial indicators, human resources, and performance with research centers in the world.   690 Neuroscience Chronic administration of bacterial lipopolysaccharide (LPS) produces a suitable model of learning and memory deficits in laboratory rats Dalvand Ismaeil Choopani Samira Hamidian Reza Sayyah Mohammad Hasanzadeh Leila Gholami Pourbadi Hamid Department of Physiology and Pharmacology, Pasteur Institute of Iran, Tehran, Iran Department of Physiology and Pharmacology, Pasteur Institute of Iran, Tehran, Iran Department of Physiology and Pharmacology, Pasteur Institute of Iran, Tehran, Iran Department of Physiology and Pharmacology, Pasteur Institute of Iran, Tehran, Iran Department of Physiology and Pharmacology, Pasteur Institute of Iran, Tehran, Iran Department of Physiology and Pharmacology, Pasteur Institute of Iran, Tehran, Iran 1 12 2023 7 281 291 08 04 2024 31 12 2023 Background and Aim: Inflammation within the central nervous system indirectly impacts memory and cognition. Bacterial lipopolysaccharide (LPS) serves as a significant contributor to inflammation and is utilized by researchers to simulate and investigate memory impairments such as Alzheimer's disease. Establishing a suitable model is essential for identifying new therapeutic candidates aimed at improving learning and memory. Thus, this study compares two different methods of administering LPS. Methods: Rats were divided into three groups: sham (PBS), single dose (10 µg LPS), and multiple doses (3 days × 5µg LPS). Intra-hippocampal injections were administered using a stereotaxic apparatus, followed by evaluating the subjects' performance in the Morris Water Maze and shuttle box tests. Results: The Morris Warer Maze test showed that repeated administration of LPS with a lower dose causes a delay in finding the hidden platform during learning. In the memory test, animals receiving repeated doses of LPS spent less time in the target quadrant than the control or single dose groups (p < 0.05). Also, in the shuttle box test, multiple administration of LPS decreased the delay time in entering the dark chamber and increased the time spent in the dark chamber compared to the control group or single dose (p < 0.05). Conclusion: The findings suggest that utilizing multiple doses of LPS, as opposed to a single dose, may be a more effective approach in establishing an animal model for studying learning and memory deficits.