ABSTRACT
ObjectiveTo investigate the effect of modified Renshen Wumeitang（MRWT） on the related regulatory factors of the γ-aminobutyric acid （GABA） signaling pathway in colon tissues of rats with diarrhea， and reveal the mechanism of MRWT in invigorating Qi， generating fluid， and checking diarrhea. MethodForty-eight SD immature rats were randomly divided into a blank group （n=12） and an experimental group （n=36）. The diarrhea model was induced in the experimental group by Sennae Folium combined with overstrain and improper diet for 14 days. Subsequently， the model rats were randomly divided into a model group （normal saline， 20 mL·kg-1）， a western medicine group （Medilac-Vita， 0.7 g·kg-1）， and a Chinese medicine group （MRWT， 35 g·kg-1）， with 12 rats in each group. The rats in the blank group received normal saline at 20 mL·kg-1， and those in the other groups were treated correspondingly， once a day for 7 days. The general condition， loose stool rate， and diarrhea index of the rats were observed daily. Immunohistochemistry was used to detect the optical density expression of GABA protein in the colon of rats. The content of phosphatidylinositol-3 kinase （PI3K）， protein kinase B2 （Akt2）， phosphorylated Akt （p-Akt）， and interleukin-1β （IL-1β） was determined by enzyme-linked immunosorbent assay （ELISA）. The mRNA and protein expression levels of PI3K， Akt2， and GABA type A receptor subunit β2 （GABRB2） in the colon of rats were detected by Real-time fluorescence quantitative polymerase chain reaction （Real-time PCR） and Western blot， respectively. ResultCompared with the blank group， the model group showed worsened general condition， The difference was not statistically significant of loose stool rate and diarrhea index， increased expression of GABA protein （P<0.05）， elevated expression of PI3K， Akt2， p-Akt， and IL-1β （P<0.05， P<0.01）， and up-regulated PI3K， Akt2， and GABRB2 mRNA and protein expression （P<0.01）. Compared with the model group， the western medicine group and the Chinese medicine group showed the improved general condition， decreased loose stool rate and diarrhea index （P<0.01）， and decreased content of PI3K， Akt2， p-Akt， and IL-1β （P<0.05）. The Chinese medicine group displayed decreased mRNA expression of PI3K， Akt2， and GABRB2 （P<0.05， P<0.01） and down-regulated protein expression of GABA， PI3K， and GABRB2 （P<0.05， P<0.01）. The western medicine group exhibited down-regulated mRNA expression of PI3K，Akt2，and protein of PI3K （P<0.05）. ConclusionMRWT can regulate the GABA signaling pathway， reduce Cl- flow in intestinal epithelial cells to the intestinal lumen， and improve the imbalance of colonic fluid metabolism in the colon of diarrhea rats， thereby exerting its effects of invigorating qi， generating fluid， and checking diarrhea.
ABSTRACT
Objective: To investigate the correlation between the polymorphisms of locus in the promoter region of glutamate decarboxylase 1 (GAD1) gene and γ-aminobutyric acid (GABA)A receptor β-3 gene (GABRB3) and schizophrenia (SZ) in Chinese Han population. Methods: SNaPshot genotyping technique was used to detect the polymorphisms of rs3791878 and rs3749034 in the promoter region of GAD1 and rs4906902 in the promoter region of GABRB3 in 545 SZ patients (case group) and 624 healthy controls (control group). The distribution of alleles and genotypes under different genetic models between the case group and the control group in all samples were compared by SNPstats online software. The above analysis was also performed after the subjects were stratified according to gender. The correlation of G/T risk genotype of rs3791878 with the age of the first onset of male SZ was investigated by survival analysis. Results: Under over-dominant genetic model, the distribution of G/T risk genotype of rs3791878 showed statistically difference between the male SZ cases and male controls (P=0.000), and the difference was still statistically significant after Bonferroni correction (P=0.000). However, there was no significant difference in the distribution of alleles and genotypes under different genetic models of rs3749034 and rs4906902 between the case group and the control group in all samples (P>0.05), and there was also no significant difference in the distribution of alleles between the case group and the control group after them being stratified according to gender (P>0.05). Kaplan-Meier analysis showed that there was no significant difference between the age of onset of male SZ who carried G/T genotype in rs3791878 locus and that of male SZ who did not carry it (P=0.603). Conclusion: The polymorphism of rs3791878 in the promoter region of GAD1 is significantly associated with the incidence of male SZ in Chinese Han population.
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Objective:To assess the anxiolytic effect of Chaimu Anshen granules (CMASG) and investigate its bioactive mechanism. Method:ICR mice were randomly divided into normal group, diazepam group(0.002 g·kg-1),Jieyu Anshen granules group(0.001 4 g·kg-1), high, medium, and low-dose (0.001 98,0.000 99,0.000 495 g·kg-1)Chaimu Anshen granule groups, with 20 mice in each group. To detect the anxiolytic effect of CMASG, mice were intragastrically administered for 4 weeks in the morning, and light-dark box transition test and open field test were performed once the other day. After the behavior tests, blood samples were collected. Six mice of each group were perfused with formalin through heart, and then the brains were fixed for immunohistochemistry test. Hippocampus of the other mice in each group were collected and stored in liquid nitrogen. The content of γ-aminobutyric acid(GABA)and glutamic acid(Glu)in hippocampus and blood samples were detected by enzyme-linked immunosorbent assay (ELISA), and the ratio of GABA/Glu was calculated. The expression of GABAα1 receptor was evaluated by the immunohistochemistry method. To test the hypnosis effect of CMASG, mice were administered intragastrically for 7 days. The sub-threshold dose of pentobarbital sodium in the sleep experiment was tested. Result:Compared with normal group, the light-dark box transitions test demonstrated that low-dose and medium-dose CMASG groups significantly prolonged the duration in light box(PPPPPPPPPPα1 receptor protein in hippocampus showed that the medium-dose CMASG significantly increased the expression of GABAα1 protein. The sub-threshold dose of pentobarbital sodium on sleep experiments confirmed that the medium-dose CMASG significantly increased the rate of sleep in mice. Conclusion:CMASG showed an anxiolytic effect, and its bioactive mechanism was related with the increase of GABA content, and the decrease of Glu content in hippocampus. Furthermore, it increased the expression of GABAα1 protein in hippocampus. The changes in content of GABA and Glu in peripheral blood were positively correlated with the changes in hippocampal tissues, which provided reference for clinical diagnosis. CMASG also exhibited an effect in improvement of sleep.
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Angelicae Gigantis Radix (AGR, Angelica gigas) has been used for a long time as a traditional folk medicine in Korea and oriental countries. Decursinol angelate (DCA) is structurally isomeric decursin, one of the major components of AGR. This study was performed to confirm whether DCA augments pentobarbital-induced sleeping behaviors via the activation of GABA(A)-ergic systems in animals. Oral administration of DCA (10, 25 and 50 mg/kg) markedly suppressed spontaneous locomotor activity. DCA also prolonged sleeping time, and decreased the sleep latency by pentobarbital (42 mg/kg), in a dose-dependent manner, similar to muscimol, both at the hypnotic (42 mg/kg) and sub-hypnotic (28 mg/kg) dosages. Especially, DCA increased the number of sleeping animals in the sub-hypnotic dosage. DCA (50 mg/kg, p.o.) itself modulated sleep architectures; DCA reduced the counts of sleep/wake cycles. At the same time, DCA increased total sleep time, but not non-rapid eye movement (NREM) and rapid eye movement (REM) sleep. In the molecular experiments. DCA (0.001, 0.01 and 0.1 µg/ml) increased intracellular Cl- influx level in hypothalamic primary cultured neuronal cells of rats. In addition, DCA increased the protein expression of glutamic acid decarboxylase (GAD(65/67)) and GABA(A) receptors subtypes. Taken together, these results suggest that DCA potentiates pentobarbital-induced sleeping behaviors through the activation of GABA(A)-ergic systems, and can be useful in the treatment of insomnia.
Subject(s)
Animals , Rats , Administration, Oral , Angelica , Electroencephalography , Eye Movements , Glutamate Decarboxylase , Korea , Medicine, Traditional , Motor Activity , Muscimol , Neurons , Pentobarbital , Receptors, GABA-A , Rodentia , Sleep Initiation and Maintenance Disorders , Sleep, REMABSTRACT
Objective To investigate the neuroprotective mechanisms of Cerebroprotein Hydrolysate for Injection (Ⅰ) on vascular dementia in rats.Method The rat vascular dementia model was prepared using an improved two-vessel occlusion method,and the common carotid artery was only isolated but not blocked in sham group.Rats were randomly divided into sham group,model group,Cerebroprotein Hydrolysate for Injection (Ⅰ) groups with low,medium and high dose (5,10,20 mg/kg) and Cerebroprotein Hydrolysate Injection group (Cerebrolysin,Positive drug,10 mg/kg).The drug was administered by iv injection of rat tail vein once a day for two weeks,while the same volume of saline was administered in sham and model group.At the end of administration,the plasma was collected through abdominal aorta to separate serum,and rat cortex was isolated to prepare homogenate.The levels of nerve growth factor (NGF) and insulin-like growth factor 2 (IGF-2) in serum and level of gamma-aminobutyric acid (GABA) in cortex were detected by ELISA.Level of glutamate (Glu) in cortex of VaD rats was detected by colorimetry.Results Compared with model group,levels of NGF and IGF-2 in the serum of VaD rats and level of GABA in cortex were significantly increased,while level of Glu in cortex was significantly decreased after administration of Cerebroprotein Hydrolysate for Injection (Ⅰ).The increased IGF-2 and GABA levels by Cerebroprotein Hydrolysate for Injection (Ⅰ) were significantly higher than that of Cerebrolysin at same dose.Conclusion The mechanisms underlying the increased leaming and memory ability of VaD rats by Cerebroprotein Hydrolysate for Injection (Ⅰ),are possibly related to the increased levels of NGF and IGF-2 in body and a regulation of the balance between excitatory and inhibitory amino acid neurotransmitters.
ABSTRACT
Objective To investigate the neuroprotective mechanisms of Cerebroprotein Hydrolysate for Injection (Ⅰ) on vascular dementia in rats.Method The rat vascular dementia model was prepared using an improved two-vessel occlusion method,and the common carotid artery was only isolated but not blocked in sham group.Rats were randomly divided into sham group,model group,Cerebroprotein Hydrolysate for Injection (Ⅰ) groups with low,medium and high dose (5,10,20 mg/kg) and Cerebroprotein Hydrolysate Injection group (Cerebrolysin,Positive drug,10 mg/kg).The drug was administered by iv injection of rat tail vein once a day for two weeks,while the same volume of saline was administered in sham and model group.At the end of administration,the plasma was collected through abdominal aorta to separate serum,and rat cortex was isolated to prepare homogenate.The levels of nerve growth factor (NGF) and insulin-like growth factor 2 (IGF-2) in serum and level of gamma-aminobutyric acid (GABA) in cortex were detected by ELISA.Level of glutamate (Glu) in cortex of VaD rats was detected by colorimetry.Results Compared with model group,levels of NGF and IGF-2 in the serum of VaD rats and level of GABA in cortex were significantly increased,while level of Glu in cortex was significantly decreased after administration of Cerebroprotein Hydrolysate for Injection (Ⅰ).The increased IGF-2 and GABA levels by Cerebroprotein Hydrolysate for Injection (Ⅰ) were significantly higher than that of Cerebrolysin at same dose.Conclusion The mechanisms underlying the increased leaming and memory ability of VaD rats by Cerebroprotein Hydrolysate for Injection (Ⅰ),are possibly related to the increased levels of NGF and IGF-2 in body and a regulation of the balance between excitatory and inhibitory amino acid neurotransmitters.
