• Journal of Ginseng Research
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• 제44권1호
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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.

• 생명과학회지
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• 제28권5호
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• pp.597-604
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• 2018

ADHD (Attention Deficit/Hyperactivity Disorder)은 4-17세의 아동 및 청소년의 약 10%가 겪는 흔한 신경 발달 장애이지만 그 핵심 기전이 알려져 있지 않은 가운데 관련한 여러 단백질들이 보고되어왔다. 이중 GIT1 (G-protein coupled-receptor kinase interacting protein-1)은 중추신경계에서 dendritic spine formation와 growth에 영향을 미치는 multifunctional adaptor protein으로, GIT1이 제거된 생쥐는 과잉행동, 주의력결핍 그리고 충동성을 보이는 ADHD 증상을 보이게 된다. 이 논문에서는 GIT1 유전자 변형 생쥐를 이용하여 genotype별로 신경교세포의 전달물질(gliotransmitter)을 비교 분석하는 실험을 진행하였다. 그 결과 주요 흥분성 전달물질인 glutamate는 HE (hetero)와 KO (knock-out)의 세포 내에서 WT (wildtype)보다 더 높은 농도로 존재했다. 한편, 억제성 신경전달물질인 GABA와 glycine의 경우 전반적으로 HE에서 가장 많은 함유량을 보였지만 소뇌 세포내의 경우, KO이 WT보다 많은 양을 함유한 것에 비해 대뇌 세포 내에서는 KO보다 WT의 억제성 전달물질 함유량이 높았다. 또한, glutamate와 GABA를 기준으로 흥분성/억제성 비율(excitation/inhibition ratio)을 보았을 때, 소뇌 세포 내/외 모두에서 KO이 가장 높은 수치를 보였고, 대뇌에서는 세포 내/외 모두 HE에서 가장 높은 수치를 보였다. 억제성 신경전달물질인 GABA가 KO의 대뇌 세포 외에서 가장 많은 것으로 보아 GIT1 결손을 보완하기 위해 억제성 물질을 더 많이 분비하거나 또는 과도하게 분비된 GABA를 재흡수하지 못하는 것이라 사료된다. 이는 ADHD 병리기전으로써 기능할 가능성을 제시하며 후속 연구를 통해 해당 기전에 대한 규명이 필요할 것으로 보인다.

• Journal of Ginseng Research
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• 제46권6호
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• pp.819-829
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• 2022

Background: Anxiolytic properties of Korean Red Ginseng (KRG) have been previously reported. However, the exact mechanism(s) of action remains to be elucidated. The present study investigated the effect of KRG on immobilization-induced anxiety-like behaviors in mice and explored the involvement of the serotonin and GABA systems and BDNF in the anxiolytic action. Methods: Mice were orally administered with KRG (200 mg/kg/day) for 4 weeks and immobilized once daily for 2 h. p-Chlorophenylalanine (p-CPA) was intraperitoneally injected on day 22-28, and flumazenil or bicuculline was injected on day 25-28. After behavioral evaluations, brains were dissected for biochemical analyses. Results: KRG improved immobilization-induced anxiety-like behaviors in mice, as assessed by the elevated plus maze (EPM) and marble burying tests (MBT). The anxiolytic effect of KRG was comparable to that of fluoxetine, a reference drug clinically used for anxiety disorders. A serotonin synthesis inhibitor, p-CPA, blocked the effect of KRG in the EPM and MBT, indicating the requirement of serotonin synthesis for anxiolytic action. In addition, the anxiolytic effect of KRG was inhibited by bicuculline (a GABA_A antagonist) in MBT, implying the involvement of GABA transmission. Western blotting analyses revealed that KRG upregulated the expression of tryptophan hydroxylase and GABA_A receptor in the brain, which was blocked by p-CPA. Enhanced BDNF expression by KRG in the hippocampus was also indicated to mediate the anxiolytic action of KRG in immobilized mice. Conclusion: KRG exhibited the anxiolytic effect in immobilized mice by multiple mechanisms of action, involving enhanced serotonin and GABA transmissions and BDNF expression.

• International Journal of Oral Biology
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• 제40권4호
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• pp.183-187
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• 2015

The present study was aimed to evaluate the influence of glutaraldehyde (GA) concentration on multiple electron microscopic (EM) immunostaining using pre-embedding peroxidase and post-embedding immunogold method. Influence of various concentrations of GA included in the fixative on immuoreactivity was assessed in the multiple immunostaining using antisera against anti-transient receptor potential vanilloid 1 (TRPV1) for peroxidase staining and anti-GABA for immunogold labeling in the rat trigeminal caudal nucleus. Anti-TRPV1 antiserum had specificity in pre-embedding peroxidase staining when tissues were fixed with fixative containing paraformaldehyde (PFA) alone. Immunoreactivity for TRPV1 was specific in tissues fixed with fixative containing 0.5% GA at both perfusion and postfixation steps, though the immunoreactivity was weaker than in tissues fixed with fixative containing PFA alone. Tissues fixed with fixative containing 0.5% GA at the perfusion and postfixation steps showed specific immunogold staining for GABA. The results of the present study indicate that GA concentration is critical for immunoreactivity to antigens such as TRPV1 and GABA. This study also suggests that the appropriate GA concentration is 0.5% for multiple immunostaining with peroxidase labeling for TRPV1 and immunogold labeling for GABA.

• 대한약학회:학술대회논문집
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• 대한약학회 2001년도 Proceedings of the Pharmaceutical Society of Korea
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• pp.124.3-125
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• 2001

• 한국약용작물학회:학술대회논문집
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• 한국약용작물학회 2008년도 춘계학술발표회
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• pp.152-153
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• 2008

• The Korean Journal of Pain
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• 제37권3호
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• pp.218-232
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• 2024

Background: Cynara scolymus has bioactive constituents and has been used for therapeutic actions. The present study was undertaken to investigate the mechanisms underlying pain-relieving effects of the hydroethanolic extract of C. scolymus (HECS). Methods: The antinociceptive activity of HECS was assessed through formalin and acetic acid-induced writhing tests at doses of 50, 100 and 200 mg/kg intraperitoneally. Additionally, naloxone (non-selective opioid receptors antagonist, 2 mg/kg), atropine (non-selective muscarinic receptors antagonist, 1 mg/kg), chlorpheniramine (histamine H₁-receptor antagonist, 20 mg/kg), cimetidine (histamine H₂-receptor antagonist, 12.5 mg/kg), flumazenil (GABA_A/BDZ receptor antagonist, 5 mg/kg) and cyproheptadine (serotonin receptor antagonist, 4 mg/kg) were used to determine the systems implicated in HECS-induced analgesia. Impact of HECS on locomotor activity was executed by open-field test. Determination of total phenolic content (TPC) and total flavonoid content (TFC) was done. Evaluation of antioxidant activity was conducted employing 2,2-diphenyl-1-picrylhydrazyl (DPPH) free radical scavenging assay. Results: HECS (50, 100 and 200 mg/kg) significantly indicated dose dependent antinociceptive activity against pain-related behavior induced by formalin and acetic acid (P < 0.001). Pretreatment with naloxone, atropine and flumazenil significantly reversed HECS-induced analgesia. Antinociceptive effect of HECS remained unaffected by chlorpheniramine, cimetidine and cyproheptadine. Locomotor activity was not affected by HECS. TPC and TFC of HECS were 59.49 ± 5.57 mgGAE/g dry extract and 93.39 ± 17.16 mgRE/g dry extract, respectively. DPPH free radical scavenging activity (IC₅₀) of HECS was 161.32 ± 0.03 ㎍/mL. Conclusions: HECS possesses antinociceptive activity which is mediated via opioidergic, cholinergic and GABAergic pathways.

• Biomolecules & Therapeutics
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• 제26권5호
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• pp.432-438
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• 2018

Worldwide, caffeine is among the most commonly used stimulatory substances. Unfortunately, significant caffeine consumption is associated with several adverse effects, ranging from sleep disturbances (including insomnia) to cardiovascular problems. This study investigates whether treatment with the Evodia rutaecarpa aqueous extract (ERAE) from berries and its major molecular component, evodiamine, can reduce the adverse caffeine-induced sleep-related and excitation effects. We combined measurements from the pentobarbital-induced sleep test, the open field test, and the locomotor activity test in mice that had been dosed with caffeine. We found that ERAE and evodiamine administration reduced the degree of caffeine-induced sleep disruption during the sleep test. Additionally, we found that evodiamine significantly inhibits caffeine-induced excitation during the open field test, as well as decreasing hyperlocomotion in the locomotor activity test. Additional in vitro experiments showed that caffeine administration decreased the expression of ${\gamma}$-aminobutyric acid $(GABA)_A$ receptor subunits in the mouse hypothalamus. However, evodiamine treatment significantly reversed this expression reduction. Taken together, our results demonstrate that ERAE and its major compound, evodiamine, provide an excellent candidate for the treatment or prevention of caffeine-induced sleep disturbances and excitatory states, and that the mechanism of these beneficial effects acts, at least in part, through the $GABA_A$-ergic system.

• 대한소아치과학회지
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• 제32권1호
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• pp.55-66
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• 2005

천궁의 메탄올 수용액 추출물의 진정 작용을 밝히기 위하여 staircase test를 시행하였으며, 근육이완 효과를 검색하기 위하여 rotarod test를 시행하였다. 진정 작용의 기전 구명을 위하여 GABA 및 glycine 수용체에 작용하는 약물인 picrotoxin, yohimbine, isoniazid, strychnine 등의 약물을 이용하여 발작 유도에 걸리는 시간을 비교 관찰하였다. 또한 생체에서의 안전성을 검증하기 위하여 Irwin의 행동시험 및 신경세포에 대한 독성을 평가하기 위하여 MTT test를 실시하여 다음의 결과를 얻었다. 1. Staircase 실험에서 천궁 추출물을 10mg/kg 용량으로 경구 투여했을 때 climbing수는 대조군과 차이가 없었으나, rearing수에서는 대조군에 비하여 42%의 감소를 보였다. 2. 30mg/kg의 용량으로 경구 투여했을 때, climbing수는 대조군과 차이가 없었으나, rearing수에서는 51%의 감소를 유발하였다. 3. 근육이완 작용을 탐색하기 위한 rotarod 실험에서는 대조군과 차이를 나타내지 않았다. 4. 천궁 추출물의 투여는 picrotoxin, isoniazid, yohimbine에 의한 발작에서는 큰 변화를 유발하지 않았으나, strychnine에 의한 발작 유도시간은 41%의 증가를 유발하였다. 5. 일반 행동에서도 특이한 변화를 유발하지 않았으며, 신경세포에 적용했을 때 특이한 독성 반응을 나타내지 않았다. 천궁의 메탄을 수용액 추출물은 현저한 진정 작용을 유발하며, 그 작용 기전으로서 glycine의 작용을 증가시켜 chloride ion의 세포내 유입으로 인한 억제성 신경전도를 유도한다고 사료된다. 또한 근육이완 작용도 나타나지 않았으며, 특이한 생체독성도 유발되지 않은 것으로 보아, 향후 치과임상에서 진정요법의 한 수단으로서 사용될 수 있다는 가능성을 제시한다.

• Biomolecules & Therapeutics
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• 제25권6호
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• pp.586-592
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• 2017

Sinomenium acutum has been long used in the preparations of traditional medicine in Japan, China and Korea for the treatment of various disorders including rheumatism, fever, pulmonary diseases and mood disorders. Recently, it was reported that Sinomenium acutum, has sedative and anxiolytic effects mediated by GABA-ergic systems. These experiments were performed to investigate whether sinomenine (SIN), an alkaloid derived from Sinomenium acutum enhances pentobarbital-induced sleep via ${\gamma}$-aminobutyric acid (GABA)-ergic systems, and modulates sleep architecture in mice. Oral administration of SIN (40 mg/kg) markedly reduced spontaneous locomotor activity, similar to diazepam (a benzodiazepine agonist) in mice. SIN shortened sleep latency, and increased total sleep time in a dose-dependent manner when co-administrated with pentobarbital (42 mg/kg, i.p.). SIN also increased the number of sleeping mice and total sleep time by concomitant administration with the sub-hypnotic dosage of pentobarbital (28 mg/kg, i.p.). SIN reduced the number of sleep-wake cycles, and increased total sleep time and non-rapid eye movement (NREM) sleep. In addition, SIN also increased chloride influx in the primary cultured hypothalamic neuronal cells. Furthermore, protein overexpression of glutamic acid decarboxylase ($GAD_{65/67}$) and $GABA_A$ receptor subunits by western blot were found, being activated by SIN. In conclusion, SIN augments pentobarbital-induced sleeping behaviors through $GABA_A$-ergic systems, and increased NREM sleep. It could be a candidate for the treatment of insomnia.