Electroacupuncture (EA) Speeds Up the Regulation of Hypothalamic Pituitary Adrenal Axis Dysfunction in Acute Surgical Trauma Rats: Mediated by Hypothalamic Gamma-Aminobutyric Acid (GABA)_AReceptors

Hypothalamic Corticotropin-releasing factor (CRF) directly activates the hypothalamic pituitary adrenal axis (HPA axis) during the surgical trauma induced stress response. Electroacupuncture (EA) has been demonstrated to have stress relieving effects in breast surgery, colorectal surgery, prostatectomy and craniotomy. This study was aimed to investigate the hypothesis that EA could regulate hypothalamic CRF in surgical trauma rats. In experiment one, Sprague-Dawley (SD) male rats were divided into intact, model (10% partial hepatectomy), sham EA and EA group. Rats from the Sham EA and EA group were stimulated at ST36-Zusanli and SP6-Sanyiniiao acupoints twice, 24 hours before the surgery and immediately after the surgery. Expressions of hypothalamic CRF and CRFR, GABA receptors, glutamate decarboxylase (GAD), serum adrenocorticotropic hormone (ACTH) and Corticosterone (CORT) were observed at 2, 4, 8 and 24 h after the surgery by radioimmunoassay (RIA), western blot, real-time PCR and immunohistochemistry. In the experiment two, SD male rats were divided into the intact, model, model + vehicle, model + L-838,417 EA and EA + L838,417 group. It was found that hypothalamus CRF, serum ACTH and CORT levels were increased in model group compared with the intact group, and those in the EA group decreased in comparison with the model group. Compared with the model group, hypothalamus-aminobutyric acid (GABA) receptor Aα3 mRNA and protein expressions of the EA group raised strikingly. In conclusion, EA alleviated surgical stress response by improving the GABA synthesis in hypothalamus, thus enhancing GABA receptors’ inhibitory regulation of the HPA axis dysfunction in rats with acute surgical trauma.

Evolution of neurotransmitter gamma-aminobutyric acid,glutamate and their receptors

Gamma-aminobutyric acid(GABA)and glutamate are two important amino acid neurotransmitters widely present in the nervous systems of mammals,insects,round worm,and platyhelminths,while their receptors are quite diversified across different animal phyla.However,the evolutionary mechanisms between the two conserved neurotransmitters and their diversified receptors remain elusive,and antagonistic interactions between GABA and glutamate signal transduction systems,in particular,have begun to attract significant attention.In this review,we summarize the extant results on the origin and evolution of GABA and glutamate,as well as their receptors,and analyze possible evolutionary processes and phylogenetic relationships of various GABAs and glutamate receptors.We further discuss the evolutionary history of Excitatory/Neutral Amino Acid Transporter(EAAT),a transport protein,which plays an important role in the GABA-glutamate“yin and yang”balanced regulation.Finally,based on current advances,we propose several potential directions of future research.

Hippocampal and cortical expression of gamma-aminobutyric acid transporter 1 and glial fibrillary acidic protein in pentylenetetrazol-induced chronic epileptic rats

BACKGROUND： Gamma-aminobutyric acid transporter plays an important role in gamma-aminobutyric acid metabolism, and is highly associated with epilepsy seizures. Pathologically, astrocytes release active substances that alter neuronal excitability, and it has been demonstrated that astrocytes play a role in epileptic seizures. OBJECTIVE： To observe changes in gamma-aminobutyric acid transporter 1 and glial fibrillary acidic protein expression in the hippocampus and cortex of the temporal lobe in rats with pentylenetetrazol-induced chronic epilepsy. DESIGN, TIME AND SETTING： Randomized, controlled, animal experiment was performed at the Department of Neurobiology, Third Military University of Chinese PLA between January 2006 and December 2007. MATERIALS： Pentylenetetrazol was purchased from Sigma, USA; rabbit anti-rat gammaaminobutyric acid transporter 1 and glial fibrillary acidic protein were from Chemicon, USA. METHODS： A total of 40 Sprague Dawley rats were divided into model and control groups. Rat models of chronic epilepsy were created by pentylenetetrazol kindling, and were subdivided into 3-, 7-, and 14-day kindling subgroups. MAIN OUTCOME MEASURES： Gamma-aminobutyric acid transporter 1 and glial fibrillary acidic protein expression, as well as the number of positive cells in the hippocampus and cortex of temporal lobe of rats, were determined by immunohistochemistry and Western blot analyses. RESULTS： Compared with the control group, the number of gamma-aminobutyric acid transporter 1 and glial fibrillary acidic protein -positive cells in the hippocampus and cortex of rats with pentylenetetrazol-induced epilepsy significantly increased, gamma-aminobutyric acid transporter 1 and glial fibrillary acidic protein expression increased after 3 days of kindling, reached a peak on day 7, and remained at elevated levels at day 14 （P〈 0.05）. CONCLUSION： Astrocytic activation and gamma-aminobutyric acid transporter 1 overexpression may contribute to pentylenetetrazol-induced epilepsy.

Effects of Gamma-Aminobutyric Acid on Performance of Lactating Sows during Heat Weather

[ Objective] To study the effects of gamma-aminobutyric acid （GABA） on performance of lactating sows during heat stress. [ Metbod] A total of 14 sows at the same parity and with close expected date of childbirth and similar body we：,ght were randomly divided into control group and experimental group. They were fed a common basal diet and a GABA （300 mg/kg） supplementary diet, respectively. The trial lasted for 21 d. [ Result ] The daily feed intake, lactation yield and average daily gain of piglets in the experimental group were increased by 9.4%, 28.5% and 10.7%, respectively. The backfat of lactating sows was decreased less, and the same with the weaning-oestrus interval. The survival ratio of piglets was increased by 4.5%. Compared with the control group, the content of lactose and fat increased significantly, but other components almost did not change. [Conclusien] Supplementing GABA in diet can improve the performance of lactating sows and promote the growth of piglets effectively.

Gamma-aminobutyric acid A receptor and N-methyl-D-aspartate receptor subunit expression in rat spiral ganglion neurons

BACKGROUND： Gamma-aminobutyric acid A （GABAA） and N-methyl-D-aspartate （NMDA） receptors are significant receptors in the central nervous system. An understanding of GABAA and NMDA receptor expression in spiral ganglion neurons （SGN） provides information for the functional role of these receptors in the auditory system. OBJECTIVE： To investigate mRNA expression of GABAA receptor （GABAAR） and NMDA receptor （NMDAR） subunits in the rat SGN. DESIGN, TIME AND SETTING： This in vitro, molecular biological study was performed at the Laboratory of Otolaryngology-Head and Neck Surgery, Guangxi Medical University, China from July 2007 to May 2008. MATERIALS： Reverse Transcriptase Kit and Taq DNA polymerase were purchased from Fermentas Burlington, ON, Canada; GABAAR and NMDAR primers were purchased from Shanghai Sangon, Shanghai, China. METHODS： SGN from 3-5 day postnatal Wistar rats was collected for primary cultures, mRNA expression of GABAAR and NMDAR subunits in the SGN was determined by reverse transcription polymerase chain reaction. MAIN OUTCOME MEASURES： Expression levels of GABAAR and NMDAR subunits were determined by quantitative analysis. RESULTS： GABAAR subunits （αl 6, β1 3, and y1 3） and NMDAR subunits （NR1, NR2A, NR2B, NR2C, NR2D, NR3A, and NR3B） were detected in the SGN. In α subunit genes of GABAAR, α1 and α3 expression was similar （P 〉 0.05） and greater than the other subunits. Of the β subunit genes, β1 subunit mRNA levels were greater than β2 and β3. Of the y subunit genes, y2 subunit mRNA levels were greater than y1 and y3. NR1 mRNA expression was the greatest of NMDAR subunits. CONCLUSION： GABAAR subunits （α1 6, β1-3, and y1-3） and NMDAR subunits （NR1, NR2A, NR2B, NR2C, NR2D, NR3A, and NR3B） were expressed in the rat SGN. Through comparison of GABAAR and NMDAR subunit expression, possible GABAAR combinations, as well as highly expressed subunit combinations, were estimated, which provided information for pharmacological and electrophysiological characteristics of GABAAR in the auditory system.

Gamma-aminobutyric acid A receptors in Alzheimer＇s disease： highly localized remodeling of a complex and diverse signaling pathway

Alzheimer＇s disease （AD）, the predominant form of dementia, is a chronic, incurable neurodegenerative disorder presenting with symptoms includ- ing progressive memory loss and disturbed emotional state. It has been estimated that dementia affects over 47 million people worldwide （Prince et al., 2015）, and with 60-80% of cases attributable to AD.

Verification of Lactobacillus brevis tolerance to simulated gastric juice and the potential effects of postbiotic gamma-aminobutyric acid in streptozotocin-induced diabetic mice

The therapeutic effect of gamma-aminobutyric acid(GABA)on diabetes was spread as one of the alarming epidemics worldwide.The study aims to investigate the function of Lactobacillus brevis KLDS_(1.0727) and KLDS_(1.0373) strains as glutamic acid decarboxylase 65(GAD65)carriers capable of generating GABA by comparing in vitro free and freeze-dried models and GABA intervention in vivo.PCR amplification of gad and in vitro i.e.,(growth rate,viability at different pH,bile tolerance,and survivability in simulated gastric juice)were performed.In vivo experiments were conducted in 7 groups of C57BL/6J mice.Each group was injected with streptozotocin(Cont_(STZ),INSSTZ,LAC1_(STZ),LAC_(1MFDSTZ),LAC_(2STZ),LAC_(2MFDSTZ))daily except for the control(Cont).One group was injected with insulin(INSSTZ).The body weight and hyperglycemia in the blood were assessed weekly,post-euthanasia blood plasma parameters,insulin,and histological examination were evaluated.Results indicated L.brevis strains demonstrated a great tolerance to bile and simulated gastric juice in vitro(P<0.05).Cont_(STZ) had the highest average glucose level(6.84±6.46)mmol/L while INS_(STZ) expressed dramatically decreed in glucose level and displayed a significant decline in the average of weekly blood glucose(−5.74±3.08)mmol/L.The lowest body weight(ContSTZ)was(19.30±0.25)g.Based on the blood plasma analysis,L.brevis strains improved good cholesterol properties,liver and kidney functions,where most of these parameters fall within the average the reference range and prevent the development of symptoms of type 1 diabetes in vivo.As recommended,L.brevis should be commonly distributed as a postbiotic GABA in pharmaceutical and nutritional applications.

Cognitive dysfunction in schizophrenia patients caused by downregulation of γ-aminobutyric acid receptor subunits

BACKGROUND The expression pattern of gamma aminobutyric acid(GABA)receptor subunits are commonly altered in patients with schizophrenia,which may lead to nerve excitation/inhibition problems,affecting cognition,emotion,and behavior.AIM To explore GABA receptor expression and its relationship with schizophrenia and to provide insights into more effective treatments.METHODS This case-control study enrolled 126 patients with schizophrenia treated at our hospital and 126 healthy volunteers who underwent physical examinations at our hospital during the same period.The expression levels of the GABA receptor subunits were detected using 1H-magnetic resonance spectroscopy.The recognized cognitive battery tool,the MATRICS Consensus Cognitive Battery,was used to evaluate the scores for various dimensions of cognitive function.The correlation between GABA receptor subunit downregulation and schizophrenia was also analyzed.RESULTS Significant differences in GABA receptor subunit levels were found between the case and control groups(P<0.05).A significant difference was also found between the case and control groups in terms of cognitive function measures,including attention/alertness and learning ability(P<0.05).Specifically,as the expression levels of GABRA1(α1 subunit gene),GABRB2(β2 subunit gene),GABRD(δsubunit),and GABRE(εsubunit)decreased,the severity of the patients’condition increased gradually,indicating a positive correlation between the downregulation of these 4 receptor subunits and schizophrenia(P<0.05).However,the expression levels of GABRA5(α5 subunit gene)and GABRA6(α6 subunit gene)showed no significant correlation with schizophrenia(P>0.05).CONCLUSION Downregulation of the GABA receptor subunits is positively correlated with schizophrenia.In other words,when GABA receptor subunits are downregulated in patients,cognitive impairment becomes more severe.

MicroRNA-502-3p regulates GABAergic synapse function in hippocampal neurons

Gamma-aminobutyric acid(GABA)ergic neurons,the most abundant inhibitory neurons in the human brain,have been found to be reduced in many neurological disorders,including Alzheimer's disease and Alzheimer's disease-related dementia.Our previous study identified the upregulation of microRNA-502-3p(miR-502-3p)and downregulation of GABA type A receptor subunitα-1 in Alzheimer's disease synapses.This study investigated a new molecular relationship between miR-502-3p and GABAergic synapse function.In vitro studies were perfo rmed using the mouse hippocampal neuronal cell line HT22 and miR-502-3p agomiRs and antagomiRs.In silico analysis identified multiple binding sites of miR-502-3p at GABA type A receptor subunitα-1 mRNA.Luciferase assay confirmed that miR-502-3p targets the GABA type A receptor subunitα-1 gene and suppresses the luciferase activity.Furthermore,quantitative reve rse transcription-polymerase chain reaction,miRNA in situ hybridization,immunoblotting,and immunostaining analysis confirmed that overexpression of miR-502-3p reduced the GABA type A receptor subunitα-1 level,while suppression of miR-502-3p increased the level of GABA type A receptor subunitα-1 protein.Notably,as a result of the overexpression of miR-502-3p,cell viability was found to be reduced,and the population of necrotic cells was found to be increased.The whole cell patch-clamp analysis of human-GABA receptor A-α1/β3/γ2L human embryonic kidney(HEK)recombinant cell line also showed that overexpression of miR-502-3p reduced the GABA current and overall GABA function,suggesting a negative correlation between miR-502-3p levels and GABAergic synapse function.Additionally,the levels of proteins associated with Alzheimer s disease were high with miR-502-3p overexpression and reduced with miR-502-3p suppression.The present study provides insight into the molecular mechanism of regulation of GABAergic synapses by miR-502-3p.We propose that micro-RNA,in particular miR-502-3p,could be a potential therapeutic to rget to modulate GABAergic synapse function in neurological disorders,including Alzheimer's disease and Alzheimer's diseaserelated dementia.

Sodium glutamate and gamma-aminobutyric acid affect iron metabolism in the rat caudate putamen

Glutamic acid and gamma-aminobutyric acid （GABA） influence iron content in the substantia nigra and globus pallidus, although the mechanisms of action remain unclear. The present study measured iron content and changes in divalent metal transporter 1 （DMT1） and hephaestin expression in the substantia nigra and caudate putamen, and explored the effects of GABA and glutamic acid on iron metabolism. Results demonstrated that iron content and DMT1 non iron response element [DMT1 （-IRE）] expression were significantly greater but hephaestin expression was significantly lower in the caudate putamen of the monosodium glutamate group compared with the control group. No significant difference in iron content was detected between the GABA and control groups. DMT1 （-IRE） expression was significantly reduced, but hephaestin expressiori was significantly increased in the GABA group compared with the control group. In addition, there was no significant difference in tyrosine hydroxylase expression between monosodium glutamate and GABA groups and the control group. These results suggested that glutamate affected iron metabolism in the caudate putamen by increasing DMTI（-IRE） and decreasing hephaestin expression. In addition, GABA decreased DMT1 （-IRE） expression in the caudate putamen.

Effects of Toll-like receptor 4 antagonist Eritrane on neurogenesis and gamma-aminobutyric acid glutamate balance in prefrontal lobe and hippocampus of depressive rats

Objective:To investigate the effects of Toll-like receptor 4 antagonist Eritrane on neurogenesis and gamma-aminobutyric acid glutamate balance in the prefrontal lobe and hippocampus of depressive rats.Methods:100 healthy SD rats were divided into healthy control group, blank control group and Eritram group (low dose group, medium dose group and high dose group), 20 rats in each group. The depression model of rats in blank control group and Salvia miltiorrhiza group was established. After the completion of the model, rats in the blank control group were injected with normal saline and rats in the Eritrean group were injected with high, medium and low doses of Eritrean injection respectively. After 21 days of continuous administration, the total distance of spontaneous activity, the immobility time of forced swimming, the levels of gamma-aminobutyric glutamate (GABA), glutamate (Glu) and Toll-like receptor 4 (TLR-4) protein in prefrontal lobe and hippocampal neurons were measured and compared. Pearson correlation test was used to analyze the correlation between TLR4 and GABA, Glu levels in depressive rats.Results: Compared with the healthy control group, the total spontaneous activity distance of the blank control group and Eritrean group decreased significantly, and the immobility time of forced swimming increased significantly (P<0.05). There was a significant difference between the blank control group and Eritrean group (P<0.05). The rats in Eritrean group spontaneously survived with the increase of dose. The total distance increased, while the immobility time of forced swimming decreased (P<0.05). Compared with the healthy control group, the levels of GABA in the prefrontal lobe and hippocampus of rats in blank control group and Eritrean group decreased significantly, the levels of Glu and TLR4 increased significantly, and the levels of GABA in Eritrean group were higher than those in blank control group, and the levels of Glu and TLR4 were lower than those in blank control group. TLR4 and TLR4 decreased significantly (P<0.05). Pearson correlation test showed that TLR4 was negatively correlated with GABA and positively correlated with Glu (P<0.05). Conclusion: Eritrean can reduce the effects of depression on the neurogenesis of prefrontal lobe and hippocampal neurons and the balance of gamma-aminobutyric acid and glutamate in rats. The mechanism is that Eritrean can increase the level of GABA and decrease the level of Glu by antagonizing Toll-like receptor 4, thus exerting the neuroprotective effect of prefrontal lobe and hippocampal neurons.

血清GABA、BDNF、5-HT、DA及NE在胃食管反流病中的表达及临床意义

目的探讨血清γ-氨基丁酸(GABA)、脑源性营养因子(BDNF)、5-羟色胺(5-HT)、去甲肾上腺素(NE)及多巴胺(DA)在胃食管反流病(GERD)中的表达及临床意义。方法选取100例GERD患者作为GERD组、40例同期健康体检者为对照组,GERD组患者给予药物治疗8周,根据治疗效果分为效果良好组和效果不良组;对照组于体检时、GERD组于治疗前、治疗后(8周时)取外周静脉血,检测血清GABA、BDNF、5-HT、DA、NE、胃动素(MTL)、胃泌素(GAS)、胆囊收缩素(CCK)水平;采用匹兹堡睡眠质量指数(PSQI)、汉密尔顿抑郁量表(HAMD)、焦虑自评量表(SAS)、日常生活能力量表(ADL)评分评估GERD组治疗前后、对照组体检时的睡眠质量、抑郁程度、焦虑状态、日常生活能力;比较GERD患者治疗前后、效果良好及效果不良组食管括约压力、立位返流时间比、卧位反流时间比、总反流时间;采用Pearson分析GERD患者治疗前GABA、BDNF、5-HT、NE及DA水平与患者食管括约肌压力、立位返流时间比、卧位返流时间比、总返流时间比、MTL、GAS、CCK的相关性。结果GERD组治疗后血清GABA、BDNF、5-HT、NE、DA、MTL、GAS水平高于治疗前、但低于对照组,血清CCK水平及各量表评分均低于治疗前、但高于对照组(P<0.05);GERD组患者治疗后食管括约压力高于治疗前,立位返流时间比、卧位反流时间比、总反流时间均低于治疗前(P<0.05);30例疗效不良组GERD患者血清GABA、BDNF、5-HT、NE、DA、MTL、GAS水平及食管括约肌压力均低于70例疗效良好组,血清CCK水平、各量表评分、立位返流时间比、卧位反流时间比、总反流时间比均高于疗效良好组(P<0.05);GERD患者治疗前血清BDNF、5-HT、DA水平与MTL、GAS、食管括约肌压力呈正相关,与立位返流时间比、卧位反流时间比、总反流时间比、CCK呈负相关,GABA、NE与MTL、GAS、食管括约肌压力呈正相关,NE与立位返流时间比、CCK呈负相关性,GABA与立位返流时间比、总反流时间比、CCK呈负相关,差异有统计学意义(P<0.05)。结论GABA、BDNF、5-HT、NE及DA水平与胃内激素MTL、GAS、CCK的合成分泌及胃动力相关指标关系密切,这些神经递质可能参与GERD的发生和发展过程。

A New Synthesis of 4, 4-Diaryl/Diheteroaryl-3-butenyl Derivatives of Nipecotic Acids as GABA Transporter Inhibitors

A new method for the synthesis of 4, 4-diaryl/diheteroaryl-3-butenyl derivatives of nipecotic acid as GABA transporter inhibitors is described. The key intermediates 4-tosyl-1, 1-diaryl/diheteroaryl-1-butene 10a-d were synthesized by Wittig reaction, and followed by alkylation with （R）-3-piperidinecarboxylate. The resulting N-cycloalkylated amino acid esters 11a-d were saponified and then acidified to get the target compounds 1a-d. The preliminary bioassays showed that la-d exhibited excellent inhibition of [3H]-GABA uptake in vitro of culture cells.

γ-氨基丁酸(GABA)种子引发缓解辣椒盐胁迫的效果及生理生化的变化

种子引发是提高作物生长期耐盐性的有效方法,而γ-氨基丁酸(GABA)种子引发对辣椒(Capsicum annuum)耐盐性的效果和作用机制尚不清楚。该研究以‘茂蔬360’朝天椒为材料,分析了不同浓度γ-氨基丁酸(0、1.0、2.0、4.0、6.0、8.0μmol·L^(-1))种子引发对4~6叶期100 mmol·L^(-1) NaCl胁迫下的植株生物量、渗透调节物质、抗氧化能力、光合作用系统及钾钠离子吸收的影响。结果表明:(1)种子引发能显著增加盐胁迫下辣椒植株的生物量,以6.0μmol·L^(-1) GABA引发处理的效果最佳。(2)种子引发处理增加盐胁迫下植株可溶性糖、可溶性蛋白、脯氨酸的含量,同时,O_(2)^(-)和MDA的含量下降,抗氧化酶(SOD、POD、CAT和APX)活性增强,叶片叶绿素含量增加,叶片叶绿素荧光参数受胁迫影响程度低,叶绿素荧光指标包括F_(v)′/F_(m)′、qP_Lss、QY_Lss、NPQ_Lss和Rfd均有所上升。根、茎中的K^(+)含量和K^(+)/Na^(+)比值下降。(3)灰色关联度分析表明,GABA种子引发主要通过提高POD、CAT的活性和渗透调节物质含量来缓解盐胁迫对辣椒植株的伤害。综上所述,6.0μmol·L^(-1) GABA种子引发可有效提高辣椒苗期的耐盐性,其作用机制可能是提高了盐胁迫下辣椒植株的抗氧化能力和渗透调节能力。

海洛因诱导大鼠伏隔核NMDA受体亚基对GABA表达的影响

目的:研究大鼠伏隔核(NAc)在海洛因诱导条件性位置偏爱(CPP)状态及戒断状态下,NMDA受体亚基NR2B和NR2C对GABA表达的调控,探索GABA能神经元上NR2B和NR2C亚基在海洛因诱导中的作用。方法:将SD大鼠随机分为两组,即对照组和海洛因诱导组。海洛因诱导组又分为海洛因诱导CPP状态和海洛因戒断状态。采用海洛因小剂量递增法皮下连续注射7 d,建立海洛因诱导CPP模型,再自然戒断7 d,免疫组织化学染色检测各组大鼠伏隔核GABA和NR2B、NR2C亚基的位置关系。结果:采用小剂量递增法连续注射7 d海洛因,大鼠形成了稳定的海洛因诱导CPP模型。免疫荧光染色结果可见,各组大鼠伏隔核均有GABA、NR2B和NR2C表达,且GABA与NR2B及NR2C受体均有共表达。免疫组织化学结果显示,海洛因成瘾状态大鼠伏隔核GABA表达量明显高于对照组和戒断状态,具显著性差异(P<0.01)。戒断状态大鼠GABA表达量最低。结论:GABA的表达变化受NMDA受体亚基活性的影响;NR2B亚基对GABA的调控作用可能参与了海洛因依赖的复燃。

γ-aminobutyric acid B2 receptor:A potential therapeutic target for cholangiocarcinoma in patients with diabetes mellitus

BACKGROUND The association between diabetes mellitus(DM)and the increased risk and progression of cholangiocarcinoma(CCA)has been reported with unclear underlying mechanisms.Previous studies showed thatγ-aminobutyric acid(GABA)B2 receptor(GABBR2)was upregulated in CCA cells cultured in high glucose(HG)conditions.Roles of GABA receptors in CCA progression have also been studied,but their association with DM and hyperglycemia in CCA remains unclarified.AIM To investigate the effects of hyperglycemia on GABBR2 expression and the potential use of GABBR2 as a CCA therapeutic target.METHODS CCA cells,KKU-055 and KKU-213A,were cultured in Dulbecco Modified Eagle’s Medium supplemented with 5.6 mmol/L(normal glucose,NG)or 25 mmol/L(HG)glucose and assigned as NG and HG cells,respectively.GABBR2 expression in NG and HG cells was investigated using real-time quantitative polymerase chain reaction and western blot.Expression and localization of GABBR2 in CCA cells were determined using immunocytofluorescence.GABBR2 expression in tumor tissues from CCA patients with and without DM was studied using immunohistochemistry,and the correlations of GABBR2 with the clinicopathological characteristics of patients were analyzed using univariate analysis.Effects of baclofen,a GABA-B receptor agonist,on CCA cell proliferation and clonogenicity were tested using the MTT and clonogenic assays.Phospho-kinases arrays were used to screen the affected signaling pathways after baclofen treatment,and the candidate signaling molecules were validated using the public transcriptomic data and western blot.RESULTS GABBR2 expression in CCA cells was induced by HG in a dose-and time-dependent manner.CCA tissues from patients with DM and hyperglycemia also showed a significantly higher GABBR2 expression compared with tumor tissues from those with euglycemia(P<0.01).High GABBR2 expression was significantly associated with a poorer non-papillary histological subtype but with smaller sizes of CCA tumors(P<0.05).HG cells of both tested CCA cell lines were more sensitive to baclofen treatment.Baclofen significantly suppressed the proliferation and clonogenicity of CCA cells in both NG and HG conditions(P<0.05).Phospho-kinase arrays suggested glycogen synthase kinase 3(GSK3),β-catenin,and the signal transducer and activator of transcription 3(STAT3)as candidate signaling molecules under the regulation of GABBR2,which were verified in NG and HG cells of the individual CCA cell lines.Cyclin D1 and c-Myc,the common downstream targets of GSK3/β-catenin and STAT3 involving cell proliferation,were accordingly downregulated after baclofen treatment.CONCLUSION GABBR2 is upregulated by HG and holds a promising role as a therapeutic target for CCA regardless of the glucose condition.

GABA富化处理的大米产品的成分分析与营养评价

近年来随着人们生活水平的迅速改善,人们更加关注膳食营养健康。在本研究中,为了分析经过GABA富化装置处理前后稻谷的营养成分和功能组分变化,测定了GABA、游离氨基酸、蛋白质、还原糖、抗性淀粉和总淀粉等指标的含量。经过GABA富化处理后,精米中的还原糖含量由1.53 mg/g减少到0.341 mg/g;淀粉含量由746 mg/g减少到728 mg/g;而抗性淀粉含量由1.96%增加到2.30%;GABA含量由84.1μg/g增加到111μg/g;游离氨基酸含量由1114μg/g增加到1123μg/g;必需氨基酸含量由419μg/g增加到426μg/g。总蛋白含量几乎不变,且蛋白质组成符合理想蛋白质的要求。

高含量GABA桑叶茶工艺优化研究

以γ-氨基丁酸(GABA)含量为评价指标,在单因素试验基础上采用L9(33)正交试验设计方法,研究谷氨酸钠浓度、杀青功率、杀青时间对桑叶茶GABA含量的影响,确定制作高含量GABA桑叶茶的最佳工艺。结果表明,影响GABA含量的主要因素依次为谷氨酸钠浓度、杀青时间、杀青功率;经正交试验后确定最佳工艺参数为谷氨酸钠浓度5%、杀青功率210 W、杀青时间2 min,在此工艺下GABA含量最高为10.81 mg/g,同时还提高了1-脱氧野尻霉素(DNJ)的含量。

盐胁迫下玉米幼苗ABA和GABA的积累及其相互关系

盐胁迫下玉米幼苗内源脱落酸(abscisicacid,ABA)和γ-氨基丁酸(gamma-aminobutyricacid,GABA)含量增加,且ABA的积累先于GABA.用100μmol/L外源ABA和150mmol/LNacl处理使玉米幼苗内源GABA含量增加,NaCl和ABA同时处理时,对玉米幼苗GABA积累的诱导表现加合效应.ABA生物合成抑制剂氟草酮预处理后使盐胁迫诱导的GABA含量增加幅度减小,说明在盐胁迫下ABA调节GBAA积累.NaCl和ABA处理均刺激谷氨酸脱羧酶(GAD)的活性,推测盐胁迫下ABA通过调控GAD的活性而导致GABA积累.

比色法测定茶叶GABA含量的可行性研究

应用比色法、HPLC和氨基酸自动分析仪法同时测定了六大茶类共45个样品的GABA含量,结果表明,HPLC与氨基酸分析仪测定结果基本一致,比色法和HPLC法测定的普洱茶、红茶、白茶、黄茶的结果差异极显著(P<0.01),应用比色法测得的普洱茶和黑茶中GABA含量比HPLC测定结果高,而其他茶类测定结果低,推测茶叶中含有化学物质(色素、氨基酸等)干扰比色法测定。目前比色法不能应用于茶叶GABA测定,需要进一步改进方法以排除如氨基酸、茶色素等的干扰。