• Archives of Pharmacal Research
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• 제28권9호
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• pp.995-1001
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• 2005

Morphine-induced analgesia has been shown to be antagonized by ginseng total saponins (GTS), which also inhibit the development of analgesic tolerance to and physical dependence on morphine. GTS is involved in both of these processes by inhibiting morphine-6-dehydrogenase, which catalyzes the synthesis of morphinone from morphine, and by increasing the level of hepatic glutathione, which participates in the toxicity response. Thus, the dual actions of ginseng are associated with the detoxification of morphine. In addition, the inhibitory or facilitated effects of GTS on electrically evoked contractions in guinea pig ileum (I-L-receptors) and mouse vas deferens $(\delta-receptors)$ are not mediated through opioid receptors, suggesting the involvement of non-opioid mechanisms. GTS also attenuates hyperactivity, reverse tolerance (behavioral sensitization), and conditioned place preference induced by psychotropic agents, such as methamphetamine, cocaine, and morphine. These effects of GTS may be attributed to complex pharmacological actions between dopamine receptors and a serotonergic/adenosine $A_{2A}1\delta-opioid$ receptor complex. Ginsenosides also attenuate the morphine-induced cAMP signaling pathway. Together, the results suggest that GTS may be useful in the prevention and therapy of the behavioral side effects induced by psychotropic agents.

• Biomolecules & Therapeutics
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• 제7권2호
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• pp.105-111
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• 1999

Nitric oxide (NO), a cellular messenger synthesized from L-arginine by NO synthase (NOS, EC.1.14.13.39), is considered to be a regulator of gastric secretion. In the present study, the role of NO in the regulation of exocrine secretion was investigated in rat gastric chief cells. Treatment of chief cells with carba-chol resulted in an increase in the arginine conversion to citrulline, the amount of $NO_{x}$, the release of pepsine-gen, and the level of cGMP Especially, carbachol-stimulated increase of arginine to citrulline transformation, the amount of $NO_{x}$, cGMP level and the release of pepsinogen were partially reduced by the natural NOS inhibitor, $N^{G}$-monomethyl-L-arginine (MMA) and $N^{G}$, $N^{G}$-dimethyl-L-arginine (DMA). Furthermore, MMA- and DMA-induced decrease of pepsinogen secretion showed dose-dependent patters. Activation of NOS is one of the early events in receptor-mediated cascade of reactions in gastric chief cells and NO, not completely, but partially mediates gastric secretion. Agonist-stimulated pepsinogen secretion in chief cells has been considered to be mediated in adenosine 3',5'-cyclic monophosphate pathway and/or guanosine 3', 5'-cyclic monophosphate (cGMP) pathway. Taken together, the above results suggest that partial decrease of exocrine secretion following treatment of NOS inhibitor may result from the inactivation of NOS and subsequent guano- late cyclase, and NO/cGMP pathway may play a pivotal role in exocrine secretion.

• 한방재활의학과학회지
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• 제28권3호
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• pp.13-25
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• 2018

Objectives We investigated anti-obesity effects of essential oils extracted from Cnidii Rhizoma (CR) in immature adipocytes to magnify it's clinical therapeutic usage. Methods Essential oil of CR was extracted with ethyl acetate or petroleum ether and through steam distillation, respectively. Oil red-O staining for monitoring its inhibition effect on adipogenesis and differentiation in murine 3T3-L1 adipocytes and 3-(4,5-methylthiazol-2-yl)-2,5-diphenyletetra zolium bromide (MTT) assay for cell safety were done. Also phospho-adenosine monophosphate (AMP)-activted protein kinase (P-AMPK), AMP-activated protein kinase, phospho-acetyl-CoA carboxylase (P-ACC), acetyl-CoA carboxylase, peroxisome proliferator-activated receptor-${\alpha}$ (PPAR-${\alpha}$), peroxisome proliferator-activated receptor-${\gamma}$ (PPAR-${\gamma}$) and CCAAT/enhancer binding protein ${\alpha}$ (C/EBP-${\alpha}$) expressions as obesity-related factors were measured by western blot analysis. Results Protein expressions of P-AMPK, P-ACC and PPAR-${\alpha}$ were increased in essential oils-treated adipocytes compared to those of control group, respectively. Furthermore, protein expressions of PPAR-${\gamma}$ and C/EBP-${\alpha}$ were decreased in essential oils-treated adipocytes compared to those of control group, respectively. Conclusions These results demonstrate that essential oils of CR inhibit adipogenesis and differentiation. Also they promote the oxidation of fatty acids in adipocytes. Thus, results suggest that essential oils of CR could be used as a valuable material for anti-obesity therapeutics via control of lipid metabolism.

• Journal of Periodontal and Implant Science
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• 제25권3호
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• pp.635-647
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• 1995

Human polymorphonuclear leukocytes(PMN) constitute a first line of defense against all forms of injury and microbial challenge, which share a common cell lineage with macrophage. Microbial component LPS activates macrophages to produce IL-1, MIP-1${\alpha}$, -1${\beta}$, TNF-${\alpha}$ and IL-6, etc. Those cytokines have autocrine function to the macrophages, and paracrine function to other cell such as PMN and affect them to produce some biological functions. Having a responsive homogeneous cell line, HL-60, offers us the possibility of studying extensively on the function of PMN, which were not possible previously with peripheral PMN, due to the short-lived nature and difficulty of getting a purified PMN. In the present study, I performed MIP-1 receptor binding assay using HL-60 cell and human peripheral PMN. Also, in vitro antimicrobial assay was performed using differentiated or undifferentiated HL-60 cell. Differentiation was induced by treatment with 500 M of $N^6,O^2-dibutyryl$ adenosine 3'5' cyclic monophosphate(dbcAMP) (PMN-like cell), or 20ng/ml of 12-O-tetradecanoylphorbol-13-acetate(TPA) (macrophage/monocyte-like cell). Receptors for MIP-1${\alpha}$ were identified on dbcAMP-treated HL-60 as well as peripheral PMN. However, bound radioactive MIP-1${\alpha}$ on differentiated HL-60 was much higher than that of peripheral PMN, which suggest receptor number of differentiated HL-60 cell is higher than that of peripheral PMN. Although both of TPA and dbcAMP treatment significantly enhanced antimicrobial action of HL-60 cell, dbcAMP-treated cell(PMN-like HL-60) killed S.aureus more effectively in this experiment. TPA or dbcAMP treatment significantly enhanced antimicrobial action of undifferentiated HL-60 cell. MIP-1${\alpha}$ further increased enhancing effect of TPA or dbcAMP. IL-1${\alpha}$, however, increased only dbcAMP-induced enhancing effect of antimicrobial action of HL-60 cell. These results suggest that differentiated HL-60 cell could replace peripheral PMN in analysis of various biological functions of cytokines on PMN cell.

• Nutrition Research and Practice
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• 제9권6호
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• pp.606-612
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• 2015

BACKGROUND/OBJECTIVES: Several medicinal properties of Smilax china L. have been studied including antioxidant, anti-inflammatory, and anti-cancer effects. However, the antiobesity activity and mechanism by which the water-soluble fraction of this plant mediates its effects are not clear. In the present study, we investigated the lipolytic actions of the water-soluble fraction of Smilax china L. leaf ethanol extract (wsSCLE) in 3T3-L1 adipocytes. MATERIALS/METHODS: The wsSCLE was identified by measuring the total polyphenol and flavonoid content. The wsSCLE was evaluated for its effects on cell viability, lipid accumulation, glycerol, and cyclic adenosine monophosphate (cAMP) contents. In addition, western blot analysis was used to evaluate the effects on protein kinase A (PKA), PKA substrates (PKAs), and hormone-sensitive lipase (HSL). For the lipid accumulation assay, 3T3-L1 adipocytes were treated with different doses of wsSCLE for 9 days starting 2 days post-confluence. In other cell experiments, mature 3T3-L1 adipocytes were treated for 24 h with wsSCLE. RESULTS: Results showed that treatment with wsSCLE at 0.05, 0.1, and 0.25 mg/mL had no effect on cell morphology and viability. Without evidence of toxicity, wsSCLE treatment decreased lipid accumulation compared with the untreated adipocyte controls as shown by the lower absorbance of Oil Red O stain. The wsSCLE significantly induced glycerol release and cAMP production in mature 3T3-L1 cells. Furthermore, protein levels of phosphorylated PKA, PKAs, and HSL significantly increased following wsSCLE treatment. CONCLUSION: These results demonstrate that the potential antiobesity activity of wsSCLE is at least in part due to the stimulation of cAMP-PKA-HSL signaling. In addition, the wsSCLE-stimulated lipolysis induced by the signaling is mediated via activation of the ${\beta}$-adrenergic receptor.

• 생명과학회지
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• 제28권1호
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• pp.50-57
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• 2018

Millettia erythrocalyx는 콩과(Fabaceae)에 속하는 식물로 중국, 태국, 인도 등 열대 아열대 지역에 분포하며, 항바이러스 활성을 보유하고 있다는 보고가 있으나 항산화능과 항암활성 등에 관한 연구는 보고된 바가 없다. 따라서 본 연구에서는 M. erythrocalyx의 에탄올 추출물(EEME)을 사용하여 항산화능을 측정하고, 인체간암세포주인 HepG2에 대한 항암활성과 그 분자적 기전에 관하여 분석하였다. 먼저 DPPH radical scavenging activity를 통해 분석한 결과, EEME의 $IC_{50}$는 $2.74{\mu}g/ml$로 뛰어난 항산화능을 보유하였음을 확인하였다. 또한 EEME 농도 의존적으로 HepG2 세포의 성장을 억제하였다. EEME의 HepG2 세포 사멸 효과의 기전을 분석하기 위하여 세포주기를 분석한 결과, EEME 농도의존적으로 SubG1 세포가 증가하였으며, Annexin V 염색과 DAPI 염색을 통해 apoptotic 세포 및 apoptotic body가 증가됨을 확인하였다. 또한 apoptosis 관련 단백질들의 발현변화를 분석한 결과, EEME 처리에 의해 사멸수용체인 Fas와 pro-apoptotic 단백질인 Bax의 발현이 증가되었으며, caspase-3, -8, -9가 활성화되고 최종적으로 PARP가 분해되어 apoptosis가 유도되었음을 확인하였다. 이러한 결과들로부터 EEME는 내인성 및 외인성 경로를 통한 apoptosis 유도에 의하여 HepG2 세포의 증식을 억제하는 항암활성을 보유하였음을 확인하였다.

• 생명과학회지
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• 제32권2호
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• pp.86-93
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• 2022

Lupeol은 pentacyclic triterpene의 일종으로 다양한 질병에 약리 효과가 있는 것으로 보고되어 있으나, lupeol이 포도당 흡수에 미치는 영향은 아직 보고된 바 없다. 본 연구에서 3T3-L1 지방세포에서 포도당 흡수에 대한 lupeol의 효과를 조사하였다. 그 결과, Lupeol은 3T3-L1 지방세포에서 GLUT4를 원형질막으로 이동시켜 포도당 흡수를 촉진하였으며, 이는 PI3K/AKT 및 AMPK 경로의 활성화와 관련되어 있었다. PI3K/AKT 경로에서 lupeol은 PI3K를 활성화시키는 insulin receptor substrate 1의 인산화와 AKT의 인산화를 촉진하지만 비정형 단백질 키나아제 C isoforms ζ 및 λ의 인산화는 촉진하지 않았다. 반면, lupeol은 5 'AMP-activated protein kinase의 인산화를 촉진하였고, Lupeol의 의한 AMPK의 활성화는 원형질막-GLUT4의 발현과 세포내 포도당 흡수를 증가시키는 것으로 확인되었다. 3T3-L1 지방세포에서 lupeol에 의한 포도당 흡수 효과는 PI3K 억제제인 wortmannin 및 AMPK 억제제인 Compound C에 의해 억제됨을 통해 확인하였다. 본 연구 결과는 lupeol이 3T3-L1 지방세포에서 PI3K/AKT 및 AMPK 경로를 통해 원형질막 GLUT4의 발현을 자극함으로써 인슐린 감수성을 증가시켜 포도당 흡수를 촉진할 수 있음을 제시하고 있다.

• Journal of Nutrition and Health
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• 제57권1호
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• pp.16-26
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• 2024

토종보리수 열매의 항당뇨 효과를 규명하기 위하여, 마우스를 네 군으로 나누어, 기본 식이, 고지방·고단순당 식이, 토종보리수 열매 추출물을 저농도 (0.5%) 및 고농도 (1.0%)로 첨가한 고지방·고단순당 식이를 12주간 제공하였다. 고지방·고단순당 식이를 섭취한 군은 대조군에 비해 체중 및 체중 증가량, 부고환 지방무게가 유의적으로 증가하였으나, 고농도 보리수 열매 추출물의 급여는 부고환 지방무게를 유의적으로 감소시켰다. 고지방·고단순당군은 대조군에 비해 혈당, 인슐린, HOMA-IR값이 유의적으로 증가하였으나, 고농도 보리수 열매 추출물은 인슐린 농도를 감소시켰고, 저농도 및 고농도 추출물은 혈당 및 HOMA-IR값을 감소시켰다. 고지방·고단순당군은 대조군에 비해 IRS-2 및 AMPK 단백질 발현도가 유의적으로 감소하였으나, 고농도 보리수 열매 추출물의 급여는 IRS-2 발현도를 증가시켰고, 저농도 및 고농도 보리수 열매 추출물의 급여는 AMPK 발현도를 증가시켰다. 따라서, 토종보리수 열매는 제2형 당뇨병 동물에서 인슐린 저항성을 개선시켜 고혈당 개선효과를 나타낸 것으로 나타났다. 고지방·고단순당 식이를 섭취한 마우스에서 저농도 및 고농도 토종보리수 열매 추출물의 급여는 혈청 중성지방 농도 및 간조직의 총 지질과 중성지방 함량을 감소시켰고, 고농도추출물의 급여는 혈청 콜레스테롤 농도를 감소시켜, 토종보리수 열매는 지방간과 이상지질혈증 개선효과를 나타내었다.

• Molecules and Cells
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• 제38권2호
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• pp.138-144
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• 2015

Raloxifene is a selective estrogen receptor modulator (SERM) that binds to the estrogen receptor (ER), and exhibits potent anti-tumor and autophagy-inducing effects in breast cancer cells. However, the mechanism of raloxifene-induced cell death and autophagy is not well-established. So, we analyzed mechanism underlying death and autophagy induced by raloxifene in MCF-7 breast cancer cells. Treatment with raloxifene significantly induced death in MCF-7 cells. Raloxifene accumulated GFP-LC3 puncta and increased the level of autophagic marker proteins, such as LC3-II, BECN1, and ATG12-ATG5 conjugates, indicating activated autophagy. Raloxifene also increased autophagic flux indicators, the cleavage of GFP from GFP-LC3 and only red fluorescence-positive puncta in mRFP-GFP-LC3-expressing cells. An autophagy inhibitor, 3-methyladenine (3-MA), suppressed the level of LC3-II and blocked the formation of GFP-LC3 puncta. Moreover, siRNA targeting BECN1 markedly reversed cell death and the level of LC3-II increased by raloxifene. Besides, raloxifene-induced cell death was not related to cleavage of caspases-7, -9, and PARP. These results indicate that raloxifene activates autophagy-dependent cell death but not apoptosis. Interestingly, raloxifene decreased the level of intracellular adenosine triphosphate (ATP) and activated the AMPK/ULK1 pathway. However it was not suppressed the AKT/mTOR pathway. Addition of ATP decreased the phosphorylation of AMPK as well as the accumulation of LC3-II, finally attenuating raloxifene-induced cell death. Our current study demonstrates that raloxifene induces autophagy via the activation of AMPK by sensing decreases in ATP, and that the overactivation of autophagy promotes cell death and thereby mediates the anti-cancer effects of raloxifene in breast cancer cells.

• Nutrition Research and Practice
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• 제11권2호
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• pp.121-129
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• 2017

BACKGROUND/OBJECTIVES: The study was conducted to evaluate the effects of dietary leucine supplementation on mitochondrial biogenesis and energy metabolism in the liver of normal birth weight (NBW) and intrauterine growth-retarded (IUGR) weanling piglets. MATERIALS/METHODS: A total of sixteen pairs of NBW and IUGR piglets from sixteen sows were selected according to their birth weight. At postnatal day 14, all piglets were weaned and fed either a control diet or a leucine-supplemented diet for 21 d. Thereafter, a $2{\times}2$ factorial experimental design was used. Each treatment consisted of eight replications with one piglet per replication. RESULTS: Compared with NBW piglets, IUGR piglets had a decreased (P < 0.05) hepatic adenosine triphosphate (ATP) content. Also, IUGR piglets exhibited reductions (P < 0.05) in the activities of hepatic mitochondrial pyruvate dehydrogenase (PDH), citrate synthase (CS), ${\alpha}$-ketoglutarate dehydrogenase (${\alpha}$-KGDH), malate dehydrogenase (MDH), and complexes I and V, along with decreases (P < 0.05) in the concentration of mitochondrial DNA (mtDNA) and the protein expression of hepatic peroxisome proliferator-activated receptor-${\gamma}$ coactivator $1{\alpha}$ (PGC-$1{\alpha}$). Dietary leucine supplementation increased (P < 0.05) the content of ATP, and the activities of CS, ${\alpha}$-KGDH, MDH, and complex V in the liver of piglets. Furthermore, compared to those fed a control diet, piglets given a leucine-supplemented diet exhibited increases (P < 0.05) in the mtDNA content and in the mRNA expressions of sirtuin 1, PGC-$1{\alpha}$, nuclear respiratory factor 1, mitochondrial transcription factor A, and ATP synthase, $H^+$ transporting, mitochondrial F1 complex, ${\beta}$ polypeptide in liver. CONCLUSIONS: Dietary leucine supplementation may exert beneficial effects on mitochondrial biogenesis and energy metabolism in NBW and IUGR weanling piglets.