• 한국약용작물학회지
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• 제13권5호
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• pp.219-226
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• 2005

Sanguisorbae radix (SR) from Sanguisorba officinalis L. (Losaceae) is widely used in Korea and China due to its various pharmacological activity. The present study aims to investigate the effect of the methanol extract of SR on amyloid ${\beta}$ Protein(25-35) $(A{\beta}\;(25-35))$, a synthetic 25-35 amyloid peptide, -induced neurotoxicity using cultured rat cortical neurons. SR, over a concentration range of $10-50\;{\mu}g/ml$, inhibited the $A{\beta}$ (25-35) $(10\;{\mu}M)-induced$ neuronal cell death, as assessed by a 3-[4,5-dimethylthiazole-2-yl]-2,5-diphenyl-tetrazolium bromide (MTT) assay and the number of apoptotic nuclei, evidenced by Hoechst 33342 staining. Pretreatment of SR $(50\;{\mu}g/ml)$ inhibited $10\;{\mu}M\;A{\beta}$ (25-35)-induced} elevation of cytosolic calcium concentration $([Ca^{2+}]c)$, which was measured by a fluorescent dye, fluo-4 AM. SR $(10\;and\;50\;{\mu}g/ml)$ inhibited glutamate release into medium induced by $10\;{\mu}M\;A{\beta}(25-35)$, which was measured by HPLC, and generation of reactive oxygen species. These results suggest that SR prevents $A{\beta}$ (25-35)-induced neuronal cell damage in vitro.

• 대한약리학회지
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• 제26권1호
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• pp.13-23
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• 1990

가토흉부대동백근에서 MeB와 gentian violet는 용량-의존성 수축반응을 일으켰으나 evans blue와 eosine yellowish는 전혀 수축반응을 일으키지 못하였다. MeB는 돼지 장간막 동맥에서도 용량-의존성 수축반응을 일으켰다. 양표본에서 MeB $10^{-4}$ M의 단회투여는 수축반응에 이어 이완반응이 나타나는 양상성 반응을 일으켰으나, tyramine은 지속적인 수축반응을 일으켰다. Tyramine의 수축반응은 반복적이었으나 MeB의 그것은 일차 수축반응후 $3{\sim}5$시간후까지도 반복되지 않았다. Tyramine $10^{-4}$ M의 최대 수축반응 상태에서 MeB $10^{-4}$ M의 추가투여는 현저한 추가수축반응을 일으켰으나 반대로 MeB의 최대수축반응 상태에서 tyramine의 추가투여는 그이상의 수축반응을 일으키지 못하였다. Tyramine과 MeB 수축반응은 교감신경계 악물로 소실 또는 유의하게 억제되었다. Tyramine 수축반응은 MeB 수축보다 guanethidine과 6-hydroxydopamine에 더 예민한 반면, $Ca^{2+}-free$ PSS와 reserpine에 대하여는 MeB 수축반응이 tyramine 수축보다 더 예민하였고 prazosin하에서는 두 수축반응이 비슷하게 억제되었다. MeB 수축반응은 6-hydroxydopamine으로 유의하게 억제는 되었으나 소실되지 않았고, MeB 수축반응 관찰후에는 tyramine 뿐만아니라 6-hydroxydopamine의 수축반응도 소실되었다. 이상의 성적은 가토흉부대동맥과 돼지 장간막동맥에서 MeB 수축반응은 부분적으로 세포외 calcium 의존성이고 adrenaline성 신경발단으로부터의 norepinephrine유리에 기인하며, MeB의 norepinephrine유리 및 고갈작용이 tyramine 또는 6-hydroxydopamine의 작용보다 더 강력함을 시사하고 있다.

• The Korean Journal of Physiology and Pharmacology
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• 제19권1호
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• pp.51-57
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• 2015

The etiology of periodontal disease is multifactorial. Exogenous stimuli such as bacterial pathogens can interact with toll-like receptors to activate intracellular calcium signaling in gingival epithelium and other tissues. The triggering of calcium signaling induces the secretion of pro-inflammatory cytokines such as interleukin-8 as part of the inflammatory response; however, the exact mechanism of calcium signaling induced by bacterial toxins when gingival epithelial cells are exposed to pathogens is unclear. Here, we investigate calcium signaling induced by bacteria and expression of inflammatory cytokines in human gingival epithelial cells. We found that peptidoglycan, a constituent of grampositive bacteria and an agonist of toll-like receptor 2, increases intracellular calcium in a concentration-dependent manner. Peptidoglycan-induced calcium signaling was abolished by treatment with blockers of phospholipase C (U73122), inositol 1,4,5-trisphosphate receptors, indicating the release of calcium from intracellular calcium stores. Peptidoglycan-mediated interleukin-8 expression was blocked by U73122 and 1,2-bis(2-aminophenoxy)ethane-N,N,N',N'-tetraacetic acid tetrakis (acetoxymethyl ester). Moreover, interleukin-8 expression was induced by thapsigargin, a selective inhibitor of the sarco/endoplasmic reticulum calcium ATPase, when thapsigargin was treated alone or co-treated with peptidoglycan. These results suggest that the gram-positive bacterial toxin peptidoglycan induces calcium signaling via the phospholipase C/inositol 1,4,5-trisphosphate pathway, and that increased interleukin-8 expression is mediated by intracellular calcium levels in human gingival epithelial cells.

• BMB Reports
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• 제38권6호
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• pp.709-716
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• 2005

Apoptosis and necrosis are distinguished by modality primarily. Here we show an apoptosis occurred instantly, induced by $300\;{\mu}M$ W-7 ((N-(6-aminohexyl)-5-chloro-1-naphthalenesulfonamide hydrochloride), inhibitor of calmodulin), which demonstrated necrotic modality. As early as 30 min after W-7 addition, apoptotic (sub-diploid) peak could be detected by fluorescence-activated cell sorter (FACS), “DNA ladders” began to emerge also at this time point, activity of caspase-3 elevated obviously within this period. Absence of mitochondrial membrane potential (MMP) reduction and cytochrome c, AIF (apoptosis inducing factor) release, verified that this rapid apoptosis did not proceed through mitochondria pathway. Activation of caspase-12 and changes of other endoplasmic reticulum (ER) located proteins ascertained that ER pathway mediated this necrosis-like apoptosis. Our findings suggest that it is not credible to judge apoptosis by modality. Elucidation of ER pathway is helpful to comprehend the pathology of diseases associated with ER stress, and may offer a new approach to the therapy of cancer and neurodegenerative diseases.

• 대한약리학회지
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• 제31권3호
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• pp.299-311
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• 1995

혈소판은 혈전기전의 중요요소로, monoamine성 신경전달물질의 대사에 있어서 신경계와 유사점을 가지고 있다. 따라서 항우울약물인 amitriptyline (AMT)과 sertraline (SRT)의 혈소판응집 억제와 이에 의한 세포내 신호전달 물질의 함량변동 및 단백인산화에 대한 영향을 chlorpromazine (CPZ)과 비교연구함으로써, 이들 약물의 혈소판응집 억제작용의 효능을 검정하고, 항 혈소판 및 항우울 작용기전의 일단을 규명하고자 하였다. SRT, CPZ 및 AMT은 thrombin (0.25 unit/ml)에 의한 혈소판응집을 억제하였으며, 각각의 IC50은 $4.37{\times}10^{-5}\;M$, $5.76{\times}10^{-5}\;M$ 및 $1.15{\times}10^{-4}\;M$이었다. 이러한 억제효과는 A23187$(1.0\;{\mu}M)$및 PMA(320 nM)에 의한 혈소판응집에 대해서도 유사하게 나타났다. thrombin은 혈소판응집과 아울러 thromboxane $B_2$ 및 $prostaglandin\;E_2$ 생성을 유의하게 증가시켰으며, 이러한 arachidonic acid 생성은 CPZ, AMT 및 SRT에 의하여 현저하게 억제되었다. CPZ, AMT 및 SRT은 cAMP 함량을 용량의존적으로 감소시켰으며, SRT, AMT $(1{\times}10^{-4}\;M)$ 및 CPZ $(3{\times}10^{-5}\;M)$은 cGMP 함량을 증가시키는 경향을 보였다. 한편, $Ins(1,4,5)P_3$ 함량은 thrombin 부하 후 10초 이내에 정점에 도달한 후 45초 이후까지 유지된다. CPZ과 AMT은 혈소판의 $Ins(1,4,5)P_3$ 함량을 현저히 증가시키며, thrombin에 의한 증가도 유의하게 증강시킨다. SRT은 혈소판의 $Ins(1,4,5)P_3$을 증가시키나, thrombin 부하 후 증강되지는 않았다. $Ins(1,4,5)P_3$ 증가에 이어서, $[Ca^{2+}]_i$은 thrombin 부하 후 20초에 최고점에 이르며, 이러한$[Ca^{2+}]_i$, 증가는 세 약물에 의하여 현저하게 억제되었다. 혈소판 단백인산화에 대해서, thrombin은 $41{\sim}43\;kDa$ 및 20kDa 단백인산화를 현저하게 증가시켰으며, 이는 AMT, SRT 및 CPZ에 의하여 억제되었다. CPZ, AMT 및 SRT 등의 세 약물은 유의한 항응집효과와 thromboxane생성억제 효과를 나타냈으며, 이들 약물에 의한 protein kinase C 활성억제 및 $Ins(1,4,5)P_3$의 함량증가는 각각 이들약물의 항응집효과 및 항우울성 작용기전과 연관될 수 있음을 시사한다.

• Journal of Ginseng Research
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• 제43권2호
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• pp.305-311
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• 2019

Background: Gintonin is a ginseng-derived exogenous ligand of the G protein-coupled lysophosphatidic acid (LPA) receptor. We previously reported that gintonin stimulates gliotransmitter release in primary cortical astrocytes. Astrocytes play key roles in the functions of neurovascular systems. Although vascular endothelial growth factor (VEGF) is known to influence the normal growth and maintenance of cranial blood vessels and the nervous system, there is little information about the effect of gintonin on VEGF regulation in primary astrocytes, under normal and hypoxic conditions. Methods: Using primary cortical astrocytes of mice, the effects of gintonin on the release, expression, and distribution of VEGF were examined. We further investigated whether the gintonin-mediated VEGF release protects astrocytes from hypoxia. Results: Gintonin administration stimulated the release and expression of VEGF from astrocytes in a concentration- and time-dependent manner. The gintonin-mediated increase in the release of VEGF was inhibited by the LPA1/3 receptor antagonist, Ki16425; phospholipase C inhibitor, U73122; inositol 1,4,5- triphosphate receptor antagonist, 2-APB; and intracellular $Ca^{2+}$ chelator, BAPTA. Hypoxia further stimulated astrocytic VEGF release. Gintonin treatment stimulated additional VEGF release and restored cell viability that had decreased due to hypoxia, via the VEGF receptor pathway. Altogether, the regulation of VEGF release and expression and astrocytic protection mediated by gintonin under hypoxia are achieved via the LPA receptor-VEGF signaling pathways. Conclusion: The present study shows that the gintonin-mediated regulation of VEGF in cortical astrocytes might be neuroprotective against hypoxic insults and could explain the molecular basis of the beneficial effects of ginseng on the central nervous system.

• 대한치과교정학회지
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• 제25권6호
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• pp.705-714
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• 1995

Due to the great deal of effort that has gone into the study of osteoclastic differentiation and activation over the last few decades, the mechanisms of these two events have been discovered gradually. Nitric oxide($NO^-$), which is produced from arginine by a nitric oxide synthase, opened up a new area of biological research. Recently, it has been reported that $NO^-$ is produced by osteoblasts stimulated by lipopolysaccharide and several other cytokines. In this study, the effect of sodium nitroprusside(SNP), a donor of nitric oxide($NO^-$), on osteoclast-like cell formation and on mature osteoclast function was examined. To determine the mechanism of the inhibitory effects of SNP decreased not only the basal $^{45}Ca$ release but also thee bone resorption induced by PTH and 1,25-dihydroxyvitamin $D_3\;(1,25[OH]_{2}D_3)$. The inhibitory effect of SNP on bone resorption induced by PTH appeared 2 dyas after treatment, whereas SNP effect on inhibiting bone resorption induced by $1,25[OH]_{2}D_3$ appeared at the thhird days. When chicken and rat osteeoclasts were cultured on dentin slices, treatment of $300{\mu}M$ SNP resulted in a significant decrease in dentin resorption by osteoclasts in terms of total resolution area and average individual area. We also examind the effect of SNP on formation of osteoclast-like cells that is TRAP-positive multinucleated cells from chicken and rat bone marrow cells in the presence or absence of $10^{-8}\;M\;1,25[OH]_{2}D_3$. The addition of $300{\mu}M$ SNP inhibiteed the formation of TRAP-positive multinucleated cells. The present data suggest that SNP, possibly as a $NO^-$ donor, inhibits the osteoclastic differentiation and osteoclastic activity.

• 대한화장품학회:학술대회논문집
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• 대한화장품학회 2003년도 IFSCC Conference Proceeding Book I
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• pp.13-34
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• 2003

Fructose-1, 6-diphosphate (FOP), a glycolytic metabolite is reported to ameliorate inflammation and inhibit the nitric oxide production in murine macrophages stimulated with endotoxin. It is also reported that FOP has cytoprotective effects against hypoxia or ischemia/reperfusion injury in brain and heart. In this study, we examined whether FDP has protective effects on UV-induced oxidative damage in skin cell culture system and human skin in vivo. FDP had a protective role in UVB-induced LDH release and ROS accumulation in HaCaT although it did not show direct radical scavenging effect in the experiment using 1, 1-diphenyl-2-picrylhydrazyl (DPPH). FDP also preserved cellular GSH content after UV irradiation in HaCaT and normal human fibroblast culture system. Cellular oxidative stress induces multiple downstream signaling pathways that regulate expression of multiple gene including MMP-1 and collagen, we examined the effects of FDP on UV-induced alteration of these protein expression in fibroblast culture and human skin in vivo. The increased MMP-1 expression in fibroblast and human skin by UV irradiation was significantly decreased by FDP. FDP also prevented the UV-induced decrease of collagen expression in fibroblast and human skin. Moreover, the decreasing the intracellular levels of reducing equivalents in human fibroblast by glutathione (GSH) depletion lowered the UVA dose threshold for reduction of procollagen expression, indicating that the differences of glutathione contents define the susceptibility of fibroblasts towards UV-induced reduction of procollagen expression. FDP also preserved cellular GSH content after UV irradiation, indicating that FDP has protective effects on UV-induced reduction of procollagen expression, which are possibly through maintaining intracellular reducing equivalent. Based on these premises, we examined the effect of daily use of a moisturizer containing FDP on facial wrinkle in comparison with vehicle moisturizer lacking FDP. In the clinical study, FDP significantly decreased facial wrinkle compared with vehicle alone after 6 months of use. Our results suggest that FDP has anti-aging effects in skin by increasing cellular antioxidant system and preventing oxidative signal and inflammatory reaction. Therefore FDP may be useful anti-aging agent for cosmetic purpose.

• Molecules and Cells
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• 제38권9호
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• pp.796-805
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• 2015

Gintonin is a novel ginseng-derived lysophosphatidic acid (LPA) receptor ligand. Oral administration of gintonin ameliorates learning and memory dysfunctions in Alzheimer's disease (AD) animal models. The brain cholinergic system plays a key role in cognitive functions. The brains of AD patients show a reduction in acetylcholine concentration caused by cholinergic system impairments. However, little is known about the role of LPA in the cholinergic system. In this study, we used gintonin to investigate the effect of LPA receptor activation on the cholinergic system in vitro and in vivo using wild-type and AD animal models. Gintonin induced $[Ca^{2+}]_i $ transient in cultured mouse hippocampal neural progenitor cells (NPCs). Gintonin-mediated $[Ca^{2+}]_i $ transients were linked to stimulation of acetylcholine release through LPA receptor activation. Oral administration of gintonin-enriched fraction (25, 50, or 100 mg/kg, 3 weeks) significantly attenuated scopolamine-induced memory impairment. Oral administration of gintonin (25 or 50 mg/kg, 1 2 weeks) also significantly attenuated amyloid-${\beta}$ protein ($A{\beta}$)-induced cholinergic dysfunctions, such as decreased acetylcholine concentration, decreased choline acetyltransferase (ChAT) activity and immunoreactivity, and increased acetylcholine esterase (AChE) activity. In a transgenic AD mouse model, long-term oral administration of gintonin (25 or 50 mg/kg, 3 months) also attenuated AD-related cholinergic impairments. In this study, we showed that activation of G protein-coupled LPA receptors by gintonin is coupled to the regulation of cholinergic functions. Furthermore, this study showed that gintonin could be a novel agent for the restoration of cholinergic system damages due to $A{\beta}$ and could be utilized for AD prevention or therapy.

• 대한한의학회지
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• 제26권1호
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• pp.37-45
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• 2005

In the present study, we examined the effect of crude methanolic extract (CME) from Euonymus alatus (Thunb.) Sieb on arachidonic acid (AA) cascade in SKBR3 human breast cancer cell line. CME had a potent inhibitory activity of prostaglandin E2 (PGE2) release induced by A23187, a $Ca^{2+}$ ionophore. The inhibition was concentration-dependent, with the 50 value of about 5 M. CME had no inhibitory effect on A23187-induced phosphorylation of p42/p44 extracellular signal regulated kinase/mitogen-activated protein kinase or on the liberation of [14C]-AA from the cells labeled with [14C]-AA. However, CME concentration-dependently inhibited the conversion of AA to $PGE_2$ in microsomal preparations, showing its possible inhibition of cyclooxygenase (COX). In enzyme assay in vitro, CME inhibited the activities of both constitutive COX (COX­I) and inducible COX (COX-2) in a concentration-dependent manner, with the 50 values of about 0.8 and 2M, respectively. Lineweaver-Burk plot analysis indicated that CME competitively inhibited the activities of both COX-l and -2. This study is a first demonstration that CME directly inhibits COX activity.