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Effect of Saffron (Croucus Sativus L.) Extract on Central Nervous System of Male Albino Rats

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Effect of Saffron (Croucus Sativus L.) Extract on Central Nervous System of Male Albino Rats Middle-East Journal of Scientific Research 23 (4): 599-605, 2015 ISSN 1990-9233 © IDOSI Publications, 2015 DOI: 10.5829/idosi.mejsr.2015.23.04.92101 Effect of Saffron (Croucus sativus L.) Extract on Central Nervous System of Male Albino Rats Abeer M. Waggas Department of Biology (Zoology), Faculty of Girls Education, Scientific Department, (King Abdulaziz University) (Department of Biology -University of Jeddah) Saudi Arabia Abstract: Saffron (Croucus sativus L) has been used traditionally as a spice for flavoring and coloring preparations. The aim of the present study is to evaluate the effect of saffron on some neurotransmitter content in the different brain regions, i.e. cerebellum, brainstem, striatum, cerebral cortex, hypothalamus and hippocampus. The daily (i.p.) administration of saffron extract (60mg/kg) for four weeks and subsequence withdrawal caused a significant increase in epinephrine (E), norepinephrine (NE), dopamine (DA) and serotonin (5-HT) content in all the tested brain regions at most of the time intervals studied. This is may be in a part, due to the presence of safranal which are mediated through agonized GABAA and blocked calcium channel a result the release of neurotransmitter is decreased and the content is increased. Saffron may be useful as a herbal medicine having sedative effect. Key words: Saffron Brain Epinephrine (E) Norepinephrine (NE) Dopamine (DA) and Serotonin (5-HT) Rat INTRODUCTION Saffron is generally not toxic when ingested in culinary amounts. The lethal dose is 20 g and Croucus sativus L. stigma, commonly known as theabortifacient dose is 10 g [17]. In animal study LD50 saffron, is a perennial stemless herb of the values of saffron stigma and petal extract 1.6 and 6 g/kg Iridaceaefamilythat is widely cultivated in Iran and other (intraperitonally injection), respectively in mice[18]. In countries such as India, China, Spain, Italy and Greece some reports doses between 1.2 and 2 g showed nausea [1-3]. Commercial Saffron comprise the dried red stigma and followed vomiting, diarrhea and bleeding[17]. with a small portion of the yellowish style [4] and is added Saffron widely used as spice and as coloring and to food for its color and taste [5]. Phytochemical flavoring agent in preparation of various food and researches have revealed that the color is mainly due to cosmetics in many countries specially in Saudi Arabia, it's the degraded carotenoid compounds, crocin and commonly known as zaafaran. However, few studies have crocetin[6], the flavor comes from the carotenoid oxidation been published in about their effect on central nervous products, mainly safranal[7]. system. Therefore, the present work aimed to examine the Saffron is used in traditional medicine for awide chronic administration of saffron extract on epinephrine range of indications including cramps, asthma and (E), norepinephrine (NE), dopamine (DA) and serotonin depression [8]. In modern pharmacological studies, (5-HT) contents in different brain regions (Cerebellum, saffron or its active constituents has demonstrated anti- brainstem, striatum, cerebral cortex, hypothalamus and convulsant[9], anti-depressant [10], anti-tumour activities hippocampus) of male albino rat, since these [11-14]. It is beneficial in cardiac disease [15] and decrease neurotransmitters are known to play an important role in injury in kidney and brain tissues[16]. brain function. Corresponding Author: Abeer M. Waggas, Department of Biology (Zoology), Faculty of Girls Education, Scientific Department, (King Abdulaziz University) (Department of Biology - University of Jeddah) Saudi Arabia. 599 Middle-East J. Sci. Res., 23 (4): 599-605, 2015 MATERIALS AND METHODS (5-HT) were extracted and estimated according to the method of Chang [23] modified by Ciarlone[24]. The Preparation of Saffron Extract: Saffron stigmas were fluorescence was measured in Jenway 6200 obtained from open market from Jeddah (Khorsan fluorometer. province, Iran ). Dried stigmas were sliced and macerated in water for 3 days. Then the combination was filtered Statistical Analysis: Data are expressed as mean±SD. and prepared solution incorporated 100 mg/ml The changes between the means were considered concentration from dry weight of stigma [19]. significant where the value of P> 0.05. Statistical analysis was done using Student’s pairedt-test with SPSS software Animals: Experiments were performed on male albino (Version 14). rats,Rattusrattus(120-140 g), 8-10 weeks old were obtained from animal house at the center of king Fahad of medical RESULTS researchers in Jeddah (Saudi Arabia). The animals were housed under controlled environment conditions (12 h Tables 1-3 and 4 illustrated epinephrine (E), light / dark cycle) at a temperature of 25+1 C0 and humidity norepinephrine (NE), dopamine (DA) and serotonin of 60+5 % and fed standard diet and water ad libitum. Rats (5-HT) content (ìg/g fresh tissue) of brain regions were kept for one week as acclimatization period before (Cerebellum, brainstem, striatum, cerebral cortex, the start of the experiment. All rats were handled in hypothalamus and hippocampus) of saffron extract accordance with the standard guide for the care and use treated and withdrawal male albino rats respectively. of laboratory animals. Data in Table 1 shows that the injection of 60mg/kg An adequate diet and water were allowed ad libitum (i.p.) of saffron extract for thirty days caused a significant under standard conditions of light, humidity and increase in E content in cerebellum, cerebral cortex, temperature (22-25 °C). Animals were kept in hypothalamus and hippocampus after 20 days. An polypropylene cages, each including a maximum of six increased was also found in all tested brain regions after animals. The treated animals were randomly divided into 30 days. The significant increase in E content persisted two groups. for 10 days of withdrawal in: cerebral cortex, After 2 weeks of acclimatization tothe laboratory hypothalamus and hippocampus. environment, the selected animals weredivided into 2 Saffron extract at dose 60mg/kg (i.p.) caused a groups as follow; SAFFRON TREATED GROP,was significant increase in NE content after 20 and 30 days in divided into four subgroups each of 6 rats. The animals all tested areas except in brainstem and hypothalamus were daily injected (i.p.) with 60 mg/kg of saffron extract after 20 days. After the withdrawal of the extract NE [20,21] and one subgroup was decapitated after 10, 20 content returned to the normal values an all tested regions and 30 days. To examine the withdrawal effect, the except in: cerebral cortex, hypothalamus and hippocampus animals were decapitated after 10 days of stopping the (Table 2). administration of saffron extract. CONTROL GROUPwas Moreover, the treatment caused a significant increase divided as the first group but the rats were treated with in DA content in all tested brain regions after 20 and saline 30 days except in brainstem after 20 days. The significant The treated and control rats were decapitated at the increase in DA content persisted for 10 days of the designed time intervals. The brain was rapidly and withdrawal in cerebral cortex, hypothalamus and carefully excised, separated into 2 halves. Each half was hippocampus (Table3). then dissected on dry ice glass plate, according to the Results from Table 4, shows that 60 mg/kg (i.p.) of method of Glowinski and Iversen [22], into the following saffron extract produced a significant increase in 5-HT regions: cerebellum, brainstem, striatum, cerebral cortex, content in cerebral cortex after 10 days. The significant hypothalamus and hippocampus. The brain tissues were increase was also found in all tested brain regions after 20 weighed then frozen at -80 ? C until analyzed. and 30 days except in brainstem, striatum and hypothalamus after 20 days. 5-HT content returned to the Neurotransmitter Analysis: Epinephrine (E), normal level in all tested regions after the withdrawal norepinephrine (NE) and dopamine (DA) and serotonin except in:cerebellum, hypothalamus and hippocampus. 600 Middle-East J. Sci. Res., 23 (4): 599-605, 2015 Table 1: Effect of chronic administration of saffron extract (60mg/kg, i.p.) and its subsequent withdrawalon epinephrine (E)content (ì g/g) in different brain areas of male albino rat. Time of area Cerebellum Brainstem Striatum Cerebral cortex Hypothalamus Hippocampus Decapitation (day) µg/g±S.E µg/g±S.E µg/g±S.E µg/g±S.E µg/g±S.E µg/g±S.E Treatment 10 % C 322.23±10.36 800.35±16.31 711.10±13.81 123.11±1.16 701.59±15.31 611.41±8.25 T 345.50±54.36 786.32±22.30 733.36±21.01 129.36.35±1.62 723.32±16.36 632.35±14.36 7.22 -1.75 3.13 5.07 3.09 3.42 ------------------------------------------------------------------------------------------------------------------------------------------------------------------------------- 20 % C 311.61±9.11 829.81±14.25 728.31±19.34 121.11±1.61 733.33±18.81 636.31±17.31 T 375.32±11.11 888.44±13.55 780.25±20.32 140.30±1.32 816.36±18.32 717.21±10.36 20.44* 7.06 7.13 15.84* 11.32* 12.70* ------------------------------------------------------------------------------------------------------------------------------------------------------------------------------- 30 % C 322.11±11.11 843.33±20.31 742.05±16.35 122.31±1.32 748.89±16.61 615.31±11.88 T 388.54±11.25 945.32±18.64 833.33±22.21 163.36±3.25 842.65±18.18 714.35±14.25 20.62* 12.09* 12.30* 33.36* 15.60* 16.09* withdrawal 10 % C 311.81+12.11 837.31±19.58 737.15±18.33 117.13±1.00 736.66±17.31 622.16±17.33 T 337.65±15.26 861.21±18.45 785.33±17.45 148.11±1.32 815.65±13.65 685.25±15.12 8.33 2.85 6.53 16.50* 10.72* 10.14* Statistical analyses were performed between control (C=6) and treated (T=6) animals by using paired t' test.
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