• Journal of Oriental Neuropsychiatry
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• v.22 no.2
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• pp.163-176
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• 2011

Objectives : BCS(Bambusae concretio silicae) is used as a traditional medicine in Korea for the incipient stroke. Recent reports indicated that BCS has a neuroprotective effect by anti-convulsion effect. However, it's mechanism is not well studied. The purpose of this study was to investigate into the molecular mechanism of BCS for neuroprotection in normoxia of cultured rat hippocampal neurons. Methods : BCS (1.0, 2.5, 5.0, and $10.0\;{\mu}g/m{\ell}$) was added to culture media (Neurobasal supplemented with B27) on DIV 0, given a normoxia, and the cell viability was measured by typical phase-contrast images of the cultures with 1.0, 2.5, 5.0, and $10.0\;{\mu}g/m{\ell}$ on DIV 21. Effects of BCS on the expression of various synaptic proteins ($GABA_B$ R1, $GABA_B$ R2, GlyR, PSD95) were observed by immunocytochemistry showing on DIV 3, 7 and 21. Results : Typical phase-contrast images of the cultures indicated that BCS has a protective effect of rat hippocampal cells in normoxia. The BCS upregulated $GABA_B$ R1 after normoxia on DIV 7, $GABA_A$ ${\beta}2/3$ on DIV 21 and $GABA_B$ R2 on DIV 21. And the BCS downregulated PSD95 after normoxia on DIV 7. Conclusions : The present study showed evidence for the efficacy of BCS in Typical phase-contrast images, upregulation of inhibitory neurotransmitter receptors($GABA_B$ R1) and downregulation of PSD95 which eventually protected neuronal cell death in normoxia.

• Biomedical Science Letters
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• v.27 no.2
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• pp.69-76
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• 2021

The aim of this study was to evaluate gamma-aminobutyric acid (GABA)-induced erythropoietin (EPO) and EPO-receptor expression in human Hep3B cells and Sprague Dawley (SD) rats during hypoxia. Expression levels of EPO, EPO-R mRNA, Janus kinase-2 (JAK-2), vascular endothelial growth factor (VEGF), hypoxia inducible factor-1 (HIF-1), and HIF-2 in response to GABA treatment were evaluated in cell lines. SD rats were randomly divided into 5 groups of 8 rats each, and GABA was orally administered; the groups were the normal control (NC), hypoxia-exposed (G0), as well as the GABA 1 mg/100 g body weight (BW) GABA treated group (G1), 5 mg/100 g BW GABA treated group (G5), and 10 mg/100 g BW GABA treated group (G10) with hypoxia. We analyzed EPO levels and red blood cell counts in rat blood and EPO gene expression in kidney tissue. EPO and VEGF mRNA levels in Hep3B cells exposed to hypoxia were significantly increased and further increased after GABA treatment. However, the expression of EPO-R and JAK-2 mRNAs were not affected by GABA, but hypoxia-induced HIF-1 and HIF-2 mRNA expression was inhibited by GABA. In the kidney tissue of rats exposed to hypoxia, the expression level of EPO mRNA was greatly increased, but levels in the GABA treatment groups significantly decreased. EPO levels in the serum showed the same significant trend, but the red blood cell counts were not significantly different. These findings demonstrate that HIF-1 and HIF-2 activation increase EPO expression in Hep3B cells exposed to hypoxia. However HIF decreased by GABA addition and VEGF increased significantly.

• Clinical and Experimental Reproductive Medicine
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• v.48 no.1
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• pp.50-60
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• 2021

Objective: This study investigated the mRNA expression of gamma-aminobutyric acid (GABA) receptors in the sperm of oligoasthenoteratozoospermic (OAT) and teratozoospermic (TER) men compared to normozoospermic (NOR) men, as well as the relationships between GABA receptor expression and sperm parameters, fertilization rate, and embryo quality. Methods: The mRNA expression of GABA A-α1 and GABA B-R2 receptors in sperm was examined using reverse transcription-polymerase chain reaction in three groups of patients: NOR (n=32), OAT (n=22), and TER (n=45). The fertilization rate and embryo quality were assessed in 35 patients undergoing intracytoplasmic sperm injection (ICSI; 10 NOR, 10 OAT, and 15 TER men). Results: OAT men had significantly higher mRNA expression of GABA A-α1 and GABA B-R2 receptors in sperm than NOR men; however, the difference between TER and NOR men was not significant. High levels of these receptors were significantly correlated with low sperm concentration, motility, and morphology, as well as the rate of good-quality embryos (GQEs) at the cleavage stage after ICSI. Patients whose female partners had a >50% GQE rate at the cleavage stage had significantly lower levels of GABA A-α1 receptor expression than those whose partners had a ≤50% GQE rate. Conclusion: Our findings indicate that mRNA levels of GABA receptors in human sperm are correlated with poor sperm quality and associated with embryo development after ICSI treatment. The GABA A-α1 receptor in sperm has a stronger relationship with embryo quality at the cleavage stage than the GABA B-R2 receptor.

• Journal of Life Science
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• v.27 no.5
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• pp.567-574
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• 2017

This study is to isolate and identify ${\gamma}$-amino butyric acid (GABA) producing lactic acid bacteria (LAB) from Makgeolii, traditional Korean rice wine and then establish the optimal culture conditions for GABA production. Sixty four LAB from Makgeolli were isolated according to the characteristics of the shape and color of the colony grown on MRS agar plate. The GABA production of the isolated strain cultured in MRS broth contained 1% MSG (mono-sodium glutamate) were determined and evaluated by TLC and HPLC analysis. Strain B-134 was selected for highest GABA production. From the analysis of 16S rRNA and glutamate decarboxylase B (gadB) gene sequences, strain B-134 was tentatively identified as a Lactobacillus plantarum subsp. plantarum B-134. Effects of culture parameters, including glutamic acid level, culture temperature, NaCl level, and pH on GABA production were investigated for culture optimization. The optimum culture condition for GABA production by B-134 were culture temperature of $37^{\circ}C$, pH of 5.7, NaCl content of 0% (w/v) and MSG content of 3% (w/v), which produced 25 mM of GABA during cultivation time of 48 hr. From these results, strain B-134 is expected to be utilized as useful microorganisms for GABA-enriched health beneficial food.

• Journal of Life Science
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• v.23 no.1
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• pp.102-109
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• 2013

To isolate GABA-producing microorganisms, 1,500 strains were isolated from different Chungkookjang samples and screened. From these strains, 20 were selected for further analyses based on a protease and slime-producing activity test. The MC 31 strain showed the highest GABA concentration in Chungkookjang and was used in this study. MC 31 was identified as Bacillus subtilis by an API 50CHB kit and 16S rDNA sequences analysis and named as B. subtilis MC 31. B. subtilis MC 31 showed exponential growth up to 12 hours at $37^{\circ}C$ in LB broth, and it reached a stationary phase after 24 to 36 hours of incubation. B. subtilis MC 31 showed maximum GABA content at 72 hours after incubation at $40^{\circ}C$.

• Food Engineering Progress
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• v.13 no.3
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• pp.183-189
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• 2009

Lactobacillus sakei B2-16 producing high level of $\gamma$-amino butyric acid (GABA) was previously isolated from Korean traditional fermented food, Kimchi. L. sakei B2-16 converted 99.3% of mono sodium glutamate (MSG) to GABA in Lactobacilli MRS broth supplemented with 1% MSG. In order to enhance the production of GABA by L. sakei B2-16, growth parameters such as media components and concentrations of major components were evaluated. The maximum GABA concentration was obtained by a modified rice germ extract broth containing 4%(w/v) sucrose and 1%(w/v) yeast extract. L. sakei B2-16 converted 100% of MSG to GABA in modified rice germ extract broth supplemented with 7% MSG.

• Journal of mucopolysaccharidosis and rare diseases
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• v.1 no.2
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• pp.54-59
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• 2015

Purpose: Growth hormone (GH) therapy substantially improves several cognitive functions in PWS. However, the molecular mechanisms underlying the beneficial effects of GH on cognition remain unclear in PWS. In this study, we investigated the effects of recombinant human GH on the gene expression of GABAB receptor subunits and GH/insulin-like growth factor (IGF) axis genes in the brain regions of PWS-mimicking mice (Snord116del). Methods: Snord116del mice were injected subcutaneously with 1.0 mg/kg GH or saline, once daily for 7 days. The collected brain tissues were analyzed for mRNA content using quantitative PCR (qPCR) in the cerebellum, hippocampus, and cerebral cortex. Results: GH increased the mRNA expression level of the $GABA_{B1}$ receptor subunit ($GABA_{BR1}$) and IGF-1R in the cerebellum. Furthermore, a significant positive correlation was found between the level of $GABA_{BR1}$ mRNA and the expression of the IGF-1R transcript. GH also induced an increase in the mRNA expression of IGF-2 and IGF-2R in the cerebellum. Conclusion: These data indicate that GH may provide beneficial effects on cognitive function through its influences on the expression of $GABA_{BR1}$ and GH/IGF-1 axis genes in PWS patients.

• Journal of Microbiology and Biotechnology
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• v.20 no.4
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• pp.763-766
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• 2010

An efficient and simple fermentation process was developed for the production of ${\gamma}$-aminobutyric acid (GABA) by Lactobacillus sakei B2-16. When the L. sakei B2-16 was cultivated in the rice bran extracts medium containing 4% sucrose, 1% yeast extract, and 12% monosodium glutamate, the maximum GABA concentration reached 660.0 mM with 100% conversion yield, showing the 2.4- fold higher GABA concentration compared with the modified MRS medium without the rice bran extracts. The GABA production was scaled-up from a laboratory scale (5 l) to a pilot (300 l) and a plant (5,000 l) scale to investigate the application possibility of GABA production to industrial fields. The production yields at the pilot and plant scales were similar to the laboratory scale using rice bran extracts medium, which could be effective for the low-cost production of GABA.

• Microbiology and Biotechnology Letters
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• v.49 no.1
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• pp.9-17
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• 2021

A γ-aminobutyric acid (GABA)-producing microorganism was isolated from galchi (hairtail fish, Trichiurus lepturus) jeotgal, a Korean salted and fermented seafood. The G144 isolate produced GABA excessively when incubated in MRS broth containing monosodium glutamate (MSG, 3%, w/v). G144 was identified as Lactobacillus brevis through 16S rRNA and recA gene sequencing. gadB and gadC encoding glutamate decarboxylase (GAD) and glutamate/GABA antiporter, respectively, were cloned and gadB was located downstream of gadC. The operon structure of gadCB was confirmed by reverse transcription (RT)-polymerase chain reaction. gadB was overexpressed in Escherichia coli and recombinant GAD was purified and its size was 54.4 kDa as evidenced by SDS-PAGE results. Maximum GAD activity was observed at pH 5.0 and 40℃ and the activity was dependent on pyridoxal 5'-phophate. The K_m and V_max of GAD were 8.6 mM and 0.01 mM/min, respectively.

• Journal of Ginseng Research
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• v.44 no.1
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• pp.86-95
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• 2020

Background: Ginsenoside Rb1 (Rb1), one of the most abundant protopanaxadiol-type ginsenosides, exerts excellent neuroprotective effects even though it has low intracephalic exposure. Purpose: The present study aimed to elucidate the apparent contradiction between the pharmacokinetics and pharmacodynamics of Rb1 by studying the mechanisms underlying neuroprotective effects of Rb1 based on regulation of microflora. Methods: A pseudo germ-free (PGF) rat model was established, and neuroprotective effects of Rb1 were compared between conventional and PGF rats. The relative abundances of common probiotics were quantified to reveal the authentic probiotics that dominate in the neuroprotection of Rb1. The expressions of the gamma-aminobutyric acid (GABA) receptors, including GABAA receptors (α2, β2, and γ2) and GABAB receptors (1b and 2), in the normal, ischemia/reperfusion (I/R), and I/R+Rb1 rat hippocampus and striatum were assessed to reveal the neuroprotective mechanism of Rb1. Results: The results showed that microbiota plays a key role in neuroprotection of Rb1. The relative abundance of Lactobacillus helveticus (Lac.H) increased 15.26 fold after pretreatment with Rb1. I/R surgery induced effects on infarct size, neurological deficit score, and proinflammatory cytokines (IL-1β, IL-6, and TNF-α) were prevented by colonizing the rat gastrointestinal tract with Lac.H (1 × 10⁹ CFU) by gavage 15 d before I/R surgery. Both Rb1 and Lac.H upregulated expression of GABA receptors in I/R rats. Coadministration of a GABA_A receptor antagonist significantly attenuated neuroprotective effects of Rb1 and Lac.H. Conclusion: In sum, Rb1 exerts neuroprotective effects by regulating Lac.H and GABA receptors rather than through direct distribution to the target sites.