32 24765236 Iranian Society of Physiology and Pharmacology 1148 Cell, Stem Cell and Cancer Low pH preconditioned amniotic epithelial cells for stem cell therapy of cancer Niknejad Hassan b Yazdanpanah Ghasem c Kakavand Mona d Lavaie Yasaman e b Department of Tissue Engineering and Regenerative Medicine, School of Advanced Technologies in Medicine, Shahid Beheshti University of Medical Sciences, Tehran, Iran c Department of Tissue Engineering and Regenerative Medicine, School of Advanced Technologies in Medicine, Shahid Beheshti University of Medical Sciences, Tehran, Iran d Nanomedicine and Tissue Engineering Research Center, Shahid Beheshti University of Medical Sciences, Tehran, Iran e Nanomedicine and Tissue Engineering Research Center, Shahid Beheshti University of Medical Sciences, Tehran, Iran 1 2 2016 20 1 1 4 16 01 2016 13 03 2016 Amniotic epithelial cells (AECs) possess unique characteristics, which make them a suitable source for cell-based therapeutic strategies. AECs have stem cell properties with low-immunogenicity (due to expressing HLA-G molecule and absence of MHC class I and II antigens) and no ethical problems, as well as availability in sufficient numbers, which can be obtained from a placenta. We have recently shown that the AECs have anti-cancer properties due to inhibition of angiogenesis, induction of apoptosis and cell cycle arrest probably through inhibition of HSP90. Since the viability of AECs must be improved after in vivo administration in acidic microenvironment of tumor, we clarify here that low pH preconditioning of AECs would lead to more survival of implanted cells in tumor site as well as improved functional outcomes.
1156 Neurophysiology/Pharmacology A Review of Animal Models of Alzheimer's Disease: a brief insight to Pharmacologic and genetic models Salari Sajjad f Bagheri Maryam g f Psychosocial Inury Research Center, Ilam University of Medical Sceinces, Ilam, iran g Psychosocial Inury Research Center, Ilam University of Medical Sceinces, Ilam, iran 1 2 2016 20 1 5 11 06 02 2016 11 04 2016 Alzheimer's disease (AD) is the most common form of neurodegenerative disorders. Memory loss in an alert person and impairment in the function of language, attention, perception, judgment or problem solving can occur in patients with AD. However, there are some medications in order to delay the debilitating aspects of the disease; but unfortunately, scientists could not found approaches to cure this progressive problem. Hence, in order to investigate the exact mechanisms underlying the disease and to discover novel drugs that can slow the progress or alleviate the clinical symptoms of AD, producing a model which can express the most pathophysiologic and behavioral features of the disease is a desire. Nowadays, there are different animal models developed by use of pharmacologic agents and/or genetic manipulations. In this paper, we aimed to describe different animal models of AD, genetic and pharmacologic, that are mostly used by researchers. 1155 Pharmacokinetics/Dynamics Carbon nanotube-anandamide complex exhibits sustained protective effects in an in vitro model of stroke Hassanzadeh Parichehr h Arbabi Elham i Atyabi Fatemeh j Dinarvand Rassoul k h Nanotechnology Research Center, Faculty of Pharmacy, Tehran University of Medical Sciences, Tehran, Iran i Research Center for Gastroenterology and Liver Diseases, Shahid Beheshti University of Medical Sciences, Tehran, Iran j Nanotechnology Research Center, Faculty of Pharmacy, Tehran University of Medical Sciences, Tehran, Iran k Nanotechnology Research Center, Faculty of Pharmacy, Tehran University of Medical Sciences, Tehran, Iran 1 2 2016 20 1 12 23 03 02 2016 06 04 2016 Introduction: The therapeutic potential of anandamide (AEA) for the neurological disorders may be negatively affected by its short half-life or poor solubility. The superior properties of carbon nanotubes (CNTs) for controlled drug delivery, prompted us to design AEA-CNTs complex and assess its effect in in vitro model of ischemic stroke. Methods: In this experimental study, a multi-walled CNTs (MWCNTs)-AEA complex was prepared using amino-functionalized COOH-MWCNTs and characterized by Fourier transform infrared spectroscopy and transmission electron microscopy. PC12 cells in the presence of AEA (0.5, 1, 2 μg/ml), acid- or amine-modified MWCNTs, or MWCNTs-AEA complex (2, 5, 8 μg/ml) were exposed to 1 and 3 h oxygen-glucose deprivation (OGD) followed by 24 h re-oxygenation. In vitro cytotoxicity and oxidative stress were evaluated using three-way ANOVA. Results: AEA immobilization on the aminated MWCNTs was confirmed. OGD significantly reduced cell viability (P<0.001). After 3 h of OGD induction, COOH-MWCNTs showed higher cytotoxicity than other MWCNTs (P<0.05, P<0.01, P<0.001) and MWCNTs-AEA was more protective than AEA alone (P<0.05, P<0.01). OGD increased malondialdehyde (MDA) and decreased glutathione (GSH) and superoxide dismutase (SOD) (P<0.001). Following 1-h OGD, AEA dose-dependently reduced MDA (P<0.001), and elevated GSH (P<0.05, P<0.01) and SOD (P<0.05, P<0.01), but AEA was ineffective following 3-h OGD (P>0.05). MWCNTs-AEA complex was effective at both time points (MDA and GSH: P<0.01, P<0.001, SOD: P<0.05, P<0.01, P<0.001). This nanostructure was more effective than AEA following longer exposure periods to OGD insult (P<0.05, P<0.01, P<0.001). Conclusion: Aminated MWCNTs are suitable carriers for AEA and provide longer- lasting effects against OGD insult. 1138 Endocrine Physiology/Pharmacology Antihyperglycemic and antihyperlipidemic effects of hydroalcoholic extract of Melissa officinalis (Lemon Balm) in alloxan-induced diabetic rats Khodsooz Sedigheh l Moshtaghian Jamal m Eivani Mehdi n l Divison of Animal physiology, Department of Biology, Faculty of Sciences, University of Isfahan, Isfahan, Iran m Divison of Animal physiology, Department of Biology, Faculty of Science, University of Isfahan, Isfahan, Iran n Department of Animal Biology, School of Biology and Center of Excellence in Phylogeny of Living Organisms, College of Science, University of Tehran, Tehran, Iran 1 2 2016 20 1 24 30 06 10 2015 26 04 2016 Introduction: Diabetes mellitus is a metabolic disorder of the endocrine system leading to increased blood glucose concentration in the patients. As a basic treatment for managing the blood glucose level, insulin or hypoglycemic medications are used but herbal medicines are more favored. The design of this research project was to study the therapeutic effect of hydroalcoholic extract of Melissa officinalis (HEMO) in diabetic rats. Methods: Twenty-five Wistar male rats weighing 220±25 grams were distributed semi-randomly into five groups of five each. Group 1 and 2 was respectively the control and diabetic animals. Group 3, 4 and 5 were the diabetic animals treated with HEMO either at 20, 100 or 500 mg/Kg of body weight. To induce diabetic rat models, each animals received a single intraperitoneal injection of alloxan at the dose of 120 mg/Kg. All treatments with HEMO performed daily via gavage for a period of 4 weeks. Then, blood samples were collected from all animals to measure the blood glucose level, cholesterol, triglycerides, LDL and HDL. Results: The results of this study indicated significant (P<0.05) decreases in blood sugar level, cholesterol, triglycerides and LDL in diabetic rats treated with HEMO. In addition, significant (P<0.05) increase in HDL level was observed in HEMO treated diabetic rats compared with the non-treated ones. Conclusion: HEMO has significant effects on attenuating the blood sugar level, serum lipids and lipoproteins levels, whereas it improves the HDL level. These effect might be attributed to the antioxidant benefits of flavonoids which are present in HEMO. 1149 Pharmacokinetics/Dynamics The protective effects of Curcuma longa extract on oxidative stress markers in the liver induced by Adriamycin in rat Khazdair Mohammad Reza o Mohebbati Reza p Karimi Sareh Abbasnezhad Abbasali Haghshenas Milad o Pharmaceutical Research Center and Department of Physiology School of Medicine, Mashhad University of Medical Sciences, Mashhad, Iran p Department of Physiology, School of Medicine, Medical University of Mashhad, Mashhad, Iran Department of Physiology, School of Medicine, Medical University of Mashhad, Mashhad, Iran Department of Basic Sciences, Faculty of Medicine, Gonabad University of Medical Sciences, Gonabad, Iran Department of Physiology, School of Medicine, Medical University of Mashhad, Mashhad, Iran 1 2 2016 20 1 31 37 17 01 2016 02 03 2016 Introduction: The aim of the study was to investigate the effects of Curcuma longa (C. longa) extract on Adriamycin-induced hepatotoxicity in rat. Methods: Animals were divided in six groups: Control (CO), Adriamycin (ADR), Adriamycin with Vitamin C (ADR+VitC), Vitamin C (Vit C), C. longa with Adriamycin (CL+ADR) and without Adriamycin (CL-ADR). Hepatotoxicity was induced by Adriamycin 5mg/kg and rats were treated with C. longa 1000 mg/kg and Vitamin C 100 mg/kg , per day, orally for 4 weeks. Results: In the liver tissue of ADR group, Malonyldialdehyde (MDA) level was increased significantly compared to CO group, (p < 0.05). MDA level in the treatment groups, Vit C, CL+ADR and CL-ADR were increased significantly compared to ADR group (p < 0.05, for all three groups), and compared to ADR+VitC group (p < 0.01). Thiol level in ADR, ADR+VitC and CL+ADR groups were decreased compared to CO group (p < 0.001), and also thiol level in CL-ADR and Vit C were increased significantly compared to ADR group (p < 0.01 and p < 0.001 respectively). The activity of catalase in liver tissue in ADR group was lower compared to CO group (p < 0.01), though was increased in CL-ADR, ADR+VitC and Vit C groups in comparison with ADR group (p<0.05, p < 0.01 and p < 0.001, respectively). Conclusion: The results showed that chronic administration of C. longa hydroalcoholic extract in Adriamycin-induced hepatotoxic rats could decrease the oxidative stress injuries in the liver tissue. 1157 Neurophysiology/Pharmacology Kisspeptin-13 ameliorates memory impairment induced by streptozotocin in male rats via cholinergic system Pourmir Maryam Babaei Parvin Soltani Tehrani Bahram Department of Physiology, Faculty of Medicine, Guilan University of Medical Sciences, Rasht, Iran Department of Physiology, Faculty of Medicine, Guilan University of Medical Sciences, Rasht, Iran Cellular & Molecular Research Center, Guilan University of Medical Sciences, Rasht, Iran 1 2 2016 20 1 38 47 08 02 2016 16 03 2016 Introduction: Kisspeptin-13 (KP-13) is a novel endogenous factor, increases synaptic transmission and is involved in several behavioral functions such as anxiety, locomotion, epilepsy and avoidance learning. However, the role of this peptide on cognition has not been well clarified yet. Here we studied the effect of kisspeptin-13 pretreatment on spatial learning and also interaction with cholinergic and adrenergic systems. Methods: Eighty adult male Wistar rats were divided into 10 groups: saline + saline; saline + STZ; KP-13 + STZ; propranolol + STZ; prazosin + STZ; atropine + STZ; saline + KP-13 + STZ; propranolol + KP-13 + STZ; prazosin + KP-13 + STZ; atropine + KP-13+ STZ. Streptozotocin (STZ) (3mg/Kg) was administrated intracerebroventricularly (i.c.v), kisspeptin-13 was infused (1μg/2μl, i.c.v) 30 minutes before STZ and antagonists were infused (i.p) 30 minutes before kisspeptin-13. Memory performance was measured 14 days after STZ injection using Morris Water Maze (MWM) consisting of 4 blocks and one probe tests. Results: Pretreatment with kisspeptin-13 ameliorated acquisition (p = 0.001) and retrieval of memory impaired by STZ (P = 0.011). Moreover, we found that injection of atropine, but not propranolol or prazocin was able to reverse the memory enhancement caused by kisspeptin-13 (P = 0.037). Conclusion: Our findings indicate that facilitatory action of kisspeptin-13 on the spatial learning and memory in STZ-induced Alzheimer’s is mediated, at least in part, through cholinergic systems. 1131 Blood and Immune System Inhibitory effects of oxytocin on the inflammatory parameters and vascular endothelial growth factor (VEGF) in the rat air pouch model of inflammation Ghadrdan Elliyeh Najafi Moslem Mikaily Mirak Sevda Eteraf-Oskouei Tahereh Student Research Committee, Faculty of Pharmacy, Tabriz University of Medical Sciences, Tabriz, Iran Department of Pharmacology and Toxicology, Faculty of Pharmacy, Tabriz University of Medical Sciences, Tabriz, Iran Introduction In the recent decades, there have been increasing evidence that a bi-directional communication exists among the immune, endocrine and central nervous systems which plays an important role in maintaining the body's homeostasis (Jiang et al., 1998; Lawrence and Kim, 2000; Maier, 2003; Quan and Banks, 2007; Tian et al., 2012). Oxytocin is a nonapeptide neurotransmitter, hormone produced in hypothalamic nuclei (Szeto et al., 2013). It is mainly involved in uterine contraction during parturition and the milk ejection reflex during lactation (Petersson et al., 2001). Evidence for anti-inflammatory activity of Physiol Pharmacol 20 (2016) 48-56 www.phypha.ir/ppj Abstract Introduction: The aim of the present study was to evaluate the effect of oxytocin on the angiogenesis and inflammatory parameters in air pouch model of inflammation. Methods: Inflammation was induced by injection of carrageenan into pouches in male Wistar rats. Oxytocin (4.25, 8.5 and 17 μg) was administered intra pouch at the same time as the carrageenan and then for 2 consecutive days. After 72 h, the pouches fluid was collected to determine exudates volume, interleukin 1-beta (IL-1ß) and vascular endothelial growth factor (VEGF) concentrations. Then, the pouches were dissected out, weighed and the hemoglobin concentration was assessed. Results: All three doses of oxytocin (4.25, 8.5 and 17 μg) significantly decreased volume of exudates (P<0.05, P<0.01 and P<0.001, respectively) while leukocyte accumulation in the pouch fluid was diminished by 8.5 and 17 μg oxytocin. The granulation tissue weight was also markedly reduced in comparison with the control group. A significant reduction in the angiogenesis rate in oxytocin-treated rats by all doses was seen. Interestingly, there was no significant difference between the effect of oxytocin and diclofenac on the inhibition of angiogenesis, VEGF concentration and inflammatory parameters except leukocyte accumulation. In addition, administration of oxytocin (17 μg/pouch) significantly decreased IL-1ß level (47%) compared to the control group (P<0.05). Conclusion: Oxytocin has an anti-inflammatory effect and inhibits cell influx and exudation to the site of the inflammatory response. The anti-angiogenesis effect of oxytocin may be related to the local inhibition of VEGF levels as similarly shown by diclofenac. Physiology and Pharmacology Student Research Committee, Faculty of Pharmacy, Tabriz University of Medical Sciences, Tabriz, Iran Department of Pharmacology and Toxicology, Faculty of Pharmacy, Tabriz University of Medical Sciences, Tabriz, Iran 1 2 2016 20 1 48 56 14 09 2015 24 02 2016 Introduction: The aim of the present study was to evaluate the effect of oxytocin on the angiogenesis and inflammatory parameters in air pouch model of inflammation. Methods: Inflammation was induced by injection of carrageenan into pouches in male Wistar rats. Oxytocin (4.25, 8.5 and 17 μg) was administered intra pouch at the same time as the carrageenan and then for 2 consecutive days. After 72 h, the pouches fluid was collected to determine exudates volume, interleukin 1-beta (IL-1ß) and vascular endothelial growth factor (VEGF) concentrations. Then, the pouches were dissected out, weighed and the hemoglobin concentration was assessed. Results: All three doses of oxytocin (4.25, 8.5 and 17 μg) significantly decreased volume of exudates (P<0.05, P<0.01 and P<0.001, respectively) while leukocyte accumulation in the pouch fluid was diminished by 8.5 and 17 μg oxytocin. The granulation tissue weight was also markedly reduced in comparison with the control group. A significant reduction in the angiogenesis rate in oxytocin-treated rats by all doses was seen. Interestingly, there was no significant difference between the effect of oxytocin and diclofenac on the inhibition of angiogenesis, VEGF concentration and inflammatory parameters except leukocyte accumulation. In addition, administration of oxytocin (17 μg/pouch) significantly decreased IL-1ß level (47%) compared to the control group (P<0.05). Conclusion: Oxytocin has an anti-inflammatory effect and inhibits cell influx and exudation to the site of the inflammatory response. The anti-angiogenesis effect of oxytocin may be related to the local inhibition of VEGF levels as similarly shown by diclofenac. 1159 Gastrointestinal Physiology/Pharmacology Duodenal acidification stimulates gastric H2S release through upregulating mRNA expression of cystathionine gamma lyase Mard Seyyed Ali Godarzinejad Hajar Dianat Mahin Physiology Research Center (PRC), Research Center for Infectious Diseases of Digestive System, Department of Physiology, School of Medicine, Ahvaz Jundishapur University of Medical Sciences, Ahvaz, Iran Physiology Research Center (PRC), Research Center for Infectious Diseases of Digestive System, Department of Physiology, School of Medicine, Ahvaz Jundishapur University of Medical Sciences, Ahvaz, Iran Physiology Research Center (PRC), Department of Physiology, School of Medicine, Ahvaz Jundishapur University of Medical Sciences, Ahvaz, Iran 1 2 2016 20 1 57 62 13 02 2016 18 04 2016 Introduction: It has been reported the alkaline response of pancreas to duodenal acidification is partly mediated through duodenal release of H2S, but till now the effect of duodenal acidification on gastric H2S release has not been investigated. Therefore, the present study designed to evaluate the effects of duodenal acidification on gastric H2S release and level of mRNA expression of cystathionine gamma lyase (CSE). Methods: Twenty four rats were randomly assigned into 3 groups (8 in each). They were control, pH2-, and pH3-treated groups. Under anesthesia, animals underwent midline laparotomy. Neutral isotonic saline or acidic isotonic solutions (pH=2 or 3) were injected in the duodenum 1 cm just below the pyloric sphincter. Ninety minutes after beginning the experiment, animals were sacrificed, stomachs ligated at lower esophageal sphincter and 2 ml saline infused in the stomach through pylorus and then gastric content was drained for measuring the pH. Two samples of gastric mucosal tissue were quickly snap-frozen and stored in liquid nitrogen for measuring the mucosal H2S concentration using ELISA kit and quantifying the mRNA expression of CSE by quantitative real-time PCR. Results: Duodenal acidification with acidic solution (pH=2) increased the gastric release of H2S and upregulated mRNA expression of CSE in gastric mucosa. The gastric mucous content was significantly increased in response to duodenal application of acidic solutions with pH2 and 3. Conclusion: Our findings indicated the stimulatory effect of duodenal acidification on gastric H2S release and mucous content is mediated through upregulation of CSE mRNA expression. 1144 Cell, Stem Cell and Cancer Isolation and optimization of mice skeletal muscle satellite cells using preplating method and culture media substitution Bakhtiari Nuredin Department of Biochemistry, Faculty of Basic Sciences, Young Researchers and Elite Club, Sanandaj Branch, Islamic Azad University, Sanandaj, Iran 1 2 2016 20 1 63 73 06 01 2016 13 04 2016 Introduction: Satellite cells are known as the main regenerative cell type in skeletal muscles. Our study established a modified digestion and preplating method for the isolation of slow or weak adherent cells for the enrichment of satellite cells. Low-survival rate of these primary stem cells prompted us to address whether cell culture medium substitution might change cell viability status. Methods: Skeletal muscle from 10-day-NMRI mice was gently isolated, dissected and digested by collagenase type I, IV and dispases. The isolated cells were verified by cellular (immunocytochemistry and flow-cytometry) and molecular (real-time PCR) techniques and the results were compared with sub-cultured cells (non-preplated cells) to determine the efficiency of preplating technique as a common isolating procedure of satellite cells. All data were analyzed using SPSS 16 and One Away ANOVA test. Results: The isolated cells exhibited a close gene expression pattern with satellite cells for self-renewal and fusion phases. The findings revealed that Pax7 as a self-renewal marker was expressed ~ 201.4 times higher than sub-cultured-group. Moreover, the findings obviously indicated that substitution of α-MEM to DMEM cell culture medium improves the survival rates of the cells. Conclusion: Our results recommend that preplating technique is a useful procedure for the isolation of satellite cells. In addition, it seems that substitution of culture medium paves the way for investigators to seek various therapeutic methods for skeletal muscle-related disorders such as skeletal muscle atrophy (SMA), amyotrophic lateral sclerosis (ALS), sarcopenia, diabetes and aging.