The F0/F principles were directly proportional to [Cl]i. == Western blotting of GAD and GABAAreceptors subunits == The groups were administered orally to DCA (10, 25 and 50 mg/kg) and diazepam (0. 2 mg/kg) before 1 h and 30 min respectively injected to all mice (0. 1 ml/10 g). DCA potentiates pentobarbital-induced sleeping behaviors through the activation of GABAA-ergic systems, and can be useful in the treatment of insomnia. Keywords: Decursinol angelate, Electroencephalogram, Insomnia, Pentobarbital-induced sleeping, -Aminobutyric acidity (GABA)Areceptors == INTRODUCTION == The research to find alternative compounds for the treatment of insomnia offers progressed in the past decades. Many kinds of herbs such as St, John’s wort, Kava Kava, Valerian and Passion flower have been launched in European countries [1]. Herbs because sleep aids are becoming very popular as option medicines. -Aminobutyric Acacetin acid (GABA)A-ergic transmission plays an important role in the mediation of sleep behaviors. GABA is synthesized from glutamate by glutamic acid decarboxylase (GAD) and higher levels of GABA contains in the inhibitory neurons [2]. GABA is the main inhibitory neurotransmitter in the central nervous systems (CNS) of mammalians [3]. GABA binds the ionotropic Acacetin EFNA1 and metabotropic GABA receptors on the pre- and post-synaptic nerves, resulting in hyperpolarization of neuronal cells in CNS [4, 5, 6]. Depending on the configuration of the structural subunits, it determines the pharmacological properties of the GABA receptors [7]. 1-Subunits in GABAAreceptors mediate sedation, amnesia, and ataxic effects of benzodiazepine (BZ), whereas 2-and 3-subunits are involved in their anxiolytic-like and muscle-relaxing actions. 5-Subunits are involved in at least some of the memory impairment caused by BZ [8]. The dried root of Korean Angelicae Gigantis Radix (AGR, Angelica gigas), known by the Korean name as Cham-dang-gui has been used as a medicinal herb in Korea [9]. This herb been used traditionally for the treatment of psychosomatic diseases such as excess stress, stress, depression, pain and insomnia [10, 11, 12, 13, 14]. So far, recent studies have been shown the essential oil components of AGR exhibited anxiolytic-like effects in rodents tests [15]. It was reported that Japanese Angelica root extract reversed Acacetin stress-induced sleep-reduction in pentobarbital sleeping animals through the inhibition from the central noradrenergic or the activation of GABAAreceptors in mice [16]. Decursinol angelate (DCA, among coumarin derivatives from AGR, Fig. 1) has been shown similar pharmacological activities of the extract of AGR such as and anti-amyloid protein aggregation, anti-amnesic activity, anti-nociceptive activity and protective activity of neurotoxicity [17, 18, 19, 20]. Therefore , we investigated whether DCA augments pentobarbital-induced sleeping behaviorsviathe activation of GABAA-ergic systems, and the sleep architectures of electroencephalogram (EEG) induced by DCA were analyzed. These experiments have been more focused on whether DCA might be a candidate of insomnia treatment. == Fig. 1 . Chemical structure from the decursinol angelate. == == METHODS == == Components == DCA was gifted from the professor Jae-Seon Acacetin Kang who works at the College of Pharmacy, Kyungsung University of Pharmacy. Diazepam and pentobarbital sodium were purchased from Hanlim Pharm. Co., Ltd. (Seoul, Korea) and Samjin Pharm. (Seoul, Korea), respectively. Fetal bovine serum (FBS), Dulbecco’s Modified Skull cap Medium (DMEM), Neurobasal A Medium, PenicillinStreptomycin and TrypsinEDTA were obtained from GIBCO (GrandIsland, NY, USA). The Cl- sensitive fluorescence probe N-(ethoxycarbonylmethyl)-6-methoxyquinolinium bromide (MQAE) and cytosine -D-arabinofuranoside were obtained from Sigma-Aldrich Co. (St. Louis, MO, USA). Muscimol (Tocris Bioscience, Bristol, UK) and Dimethyl sulfoxide (Amresco Solon, Ohio, USA) were purchased, respectively. The specific rabbit polyclonal antibodies against GABAAreceptors subunits or GAD65/67and the corresponding conjugated anti-rabbit immunoglobulin G-horseradish peroxidase were purchased from Abcam Inc. (Cambridge, UK). Chemiluminescent HRP substrate was obtained from Millipore Co. (Billerica, MA, USA). == Animals Acacetin == Almost all animals (ICR mice weighing 20~24 g and Sprague-Dawley rats weighing 220~250 g) were purchased from Samtako (Osan, Korea). All animals were housed in acrylic cages (456023 cm) and were kept at least 1 week intended for acclimation time. The room heat was managed at 222 and at the relative humidity (50~52%) with a 12 h light/dark cycle (lights on at 7: 00) in the animal room. Food and water were available adlibitum. All the behavioral experiments were performed between 10: 00 and 17: 00. Experiments were carried out in accordance with the Principle of Laboratory Creature Care (NIH publication No . 85-23, revised 1985) and the.
