Abstract
Purpose : To investigate the effects of the Chinese compound, Songyu Anshen Fang (SYF) on levels of GABA and GABA(B) receptor proteins in insomniac rats induced by para-chlorophenylalanine (PCPA). Methods : All rats were randomly separated into either a control group, insomnia group, or a SYF group (at a dose of 8.5 g/kg or 17 g/kg body weight per day). The rat model of insomnia was induced by intraperitoneal injection of PCPA, and SYF was administered intragastrically in suspension. All experimental groups were treated with a corresponding agent for one week. The levels of glutamic acid (Glu) and γ-aminobutyric acid (GABA) were determined by high performance liquid chromatography (HPLC); mRNA and protein expressions, and GABA(B) receptor levels were detected by real-time polymerase chain reaction (RT-PCR) and western blot. Results : SYF treatment with 8.5 or 17 g/kg/day decreased the levels of Glu and Glu/GABA ratios in the hypothalamus following abnormal increase by PCPA. Moreover, GABA(B) receptor, mRNA and protein expression decreased by PCPA in hypothalamus were significantly normalized by SYF. Conclusion : The study indicates that the effects of PCPA-induced insomnia can be alleviated by SYF modulation of neurotransmitter levels and the expression of GABA(B) receptor in the hypothalamus. This suggests that clinical application of SYF to treat insomnia may be feasible. Keywords : Songyu Anshen Fang, Para-chlorophenylalanine (PCPA), γ-Aminobutyric acid (GABA), GABA(B) receptor, Insomnia
Highlights
Insomnia is a widespread health problem and increasing incidence of insomnia, due to an accelerated life pace, renders insomnia research extremely important
To observe the effects of Songyu Anshen Fang (SYF) on levels of glutamic acid (Glu) and GABA in the hypothalamus of insomniac rats, the levels of Glu and GABA were determined by high performance liquid chromatography (HPLC)
real-time polymerase chain reaction (RT-PCR) results showed that after PCPA treatment, GABA(B) mRNA expression was significantly decreased (p < 0.001) in rat hypothalamus compared with the saline control group
Summary
Insomnia is a widespread health problem and increasing incidence of insomnia, due to an accelerated life pace, renders insomnia research extremely important. Insomnia is attributed at least in part to the role of neurotransmitters in regulation of sleep and wakefulness. In this study, we developed a rat model of insomnia based through the administration of PCPA. Two hours after the last day of SYF administration, the animals were deeply anesthetized by intraperitoneal injection of pentobarbital sodium (65 mg/kg) and the hypothalamus was excised and flash frozen in liquid nitrogen, followed by storage at −80 °C. In this study we investigated the effects of SYF on levels of glutamic acid, γ-aminobutyric acid and GABA(B) receptors in the hypothalamus of insomniac rats. The insomnia model was established by injection of 300 mg/kg PCPA intraperitoneally (i.p.) at 19:00 h for two consecutive days. Statistical significance between groups was examined by one-way analysis of variance (ANOVA) followed by Tukey’s test for multiple comparisons.
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