• 대한약학회:학술대회논문집
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• 대한약학회 2001년도 Proceedings of the Pharmaceutical Society of Korea
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• pp.101.2-102
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• 2001

• The Korean Journal of Physiology and Pharmacology
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• 제23권5호
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• pp.345-356
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• 2019

Docosahexaenoic acid (DHA), an omega-3-fatty acid, modulates multiple cellular functions. In this study, we addressed the effects of DHA on human umbilical vein endothelial cell calcium transient and endothelial nitric oxide synthase (eNOS) phosphorylation under control and adenosine triphosphate (ATP, $100{\mu}M$) stimulated conditions. Cells were treated for 48 h with DHA concentrations from 3 to $50{\mu}M$. Calcium transient was measured using the fluorescent dye Fura-2-AM and eNOS phosphorylation was addressed by western blot. DHA dose-dependently reduced the ATP stimulated $Ca^{2+}$-transient. This effect was preserved in the presence of BAPTA (10 and $20{\mu}M$) which chelated the intracellular calcium, but eliminated after withdrawal of extracellular calcium, application of 2-aminoethoxy-diphenylborane ($75{\mu}M$) to inhibit store-operated calcium channel or thapsigargin ($2{\mu}M$) to delete calcium store. In addition, DHA ($12{\mu}M$) increased ser1177/thr495 phosphorylation of eNOS under baseline conditions but had no significant effect on this ratio under conditions of ATP stimulation. In conclusion, DHA dose-dependently inhibited the ATP-induced calcium transient, probably via store-operated calcium channels. Furthermore, DHA changed eNOS phosphorylation suggesting activation of the enzyme. Hence, DHA may shift the regulation of eNOS away from a $Ca^{2+}$ activated mode to a preferentially controlled phosphorylation mode.

• 대한약리학회지
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• 제32권3호
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• pp.417-424
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• 1996

본 연구는 C5a에 의해 자극된 호중구에서 세포내 칼슘유리와 세포외부로부터 칼슘유입에 있어 protein kinase C와 protein tyrosine kinase의 관여 여부를 조사하였다. Protein kinase C 억제제인 staurosporine과 H-7은 C5a에 의해 자극된 호중구에서 세포내 칼슘유리를 억제하였으나, 세포막을 교차한 칼슘유입이나 세포내 칼슘농도 증가에 영향을 나타내지 않았다. C5a에 의한 세포내 칼슘유리와 칼슘유입은 protein tyrosine kinase 억제제인 genistein과 methyl-2,5-dihydroxycinnamate에 의해서 억제 되었다. ADP에 의해 야기된 세포내 칼슘농도의 증가는 genistein과 methyl-2,5-dihydroxycinnamate에 의해서 억제되었으나 staurosporine과 H-7의 영향은 받지 않았다. Genistein과 methyl-2,5-dihydroxy-cinnamate는 thapsigargin을 처리한 호중구에서 칼슘유입을 감소시켰으나 이에 대한 staurosporine 과 H-7의 효과는 나타나지 않았다. 호중구를 phorbol 12-myristate 13-acetate로 전처치하였을때 세포내 칼슘증가에 미치는 C5a의 자극 효과는 감소하였다. 이상의 결과로 부터 protein tyrosine kinase는 C5a에 의해 활성화된 호중구에서 세포내 칼슘유리와 세포막을 교차한 칼슘유입의 조절에 관여할 것으로 추정된다.

• Journal of Ginseng Research
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• 제42권2호
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• pp.165-174
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• 2018

Background: Extended endoplasmic reticulum (ER) stress may initiate apoptotic pathways in cancer cells, and ER stress has been reported to possibly increase tumor death in cancer therapy. We previously reported that caspase-8 played an important role in compound K-induced apoptosis via activation of caspase-3 directly or indirectly through Bid cleavage, cytochrome c release, and caspase-9 activation in HL-60 human leukemia cells. The mechanisms leading to apoptosis in A549 and SK-MES-1 human lung cancer cells and the role of ER stress have not yet been understood. Methods: The apoptotic effects of compound K were analyzed using flow cytometry, and the changes in protein levels were determined using Western blot analysis. The intracellular calcium levels were monitored by staining with Fura-2/AM and Fluo-3/AM. Results: Compound K-induced ER stress was confirmed through increased phosphorylation of $eIF2{\alpha}$ and protein levels of GRP78/BiP, XBP-1S, and $IRE1{\alpha}$ in human lung cancer cells. Moreover, compound-K led to the accumulation of intracellular calcium and an increase in m-calpain activities that were both significantly inhibited by pretreatment either with BAPTA-AM (an intracellular $Ca^{2+}$ chelator) or dantrolene (an RyR channel antagonist). These results were correlated with the outcome that compound K induced ER stress-related apoptosis through caspase-12, as z-ATAD-fmk (a specific inhibitor of caspase-12) partially ameliorated this effect. Interestingly, 4-PBA (ER stress inhibitor) dramatically improved the compound K-induced apoptosis. Conclusion: Cell survival and intracellular $Ca^{2+}$ homeostasis during ER stress in human lung cancer cells are important factors in the induction of the compound K-induced apoptotic pathway.

• 한국임상수의학회지
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• 제18권2호
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• pp.93-97
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• 2001

병원성 Mycoplasma hyopneumoniae strain 91-3, 비병원성 M. hyopneumoniae 그리고 M. flocculare를 돼지의 기관지섬모상피에 투여시 세포내 $Ca^{2+}$ 농도 [$Ca^{2+]}$$_{i}$ 의 변화를 본 연구에서 조사하였다. M. hyopneumoniae strain 91-3 (300-$\mu\textrm{g}$ml)를 투여시 기관지 섬모상피내의 $Ca^{2+}$가 투여전과 비교시 투여 후 250$\pm$19 nM (net increase)증가하였다 (10회 반복 47 cells). 이와는 대조적으로 비병원성 M. hyopneumoniae (300 $\mu\textrm{g}$ml) (6회 반복 18 cells)와 M. flocculare (300 $\mu\textrm{g}$ml) (8회 반복 24 cells)는 세포내 $Ca^{2+}$의 농도를 증가시키지 못하였다. 위의 결과로 병원성 M. hyopneumoniae 91-3 균주는 비병원성 mycoplasma와는 다르게 돼지의 섬모상피에서 [$Ca^{2+}$]$_{i}$ 을 유도하였으며 이러한 특성은 mycoplasma 감염증 치료에 중요한 단서를 제공할 뿐만 아니라 새로운 치료법의 개발에 유용한 스크리닝 기술에 응용될 것으로 기대된다.

• 생약학회지
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• 제44권3호
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• pp.286-290
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• 2013

This study was carried out to the determine the effects of the water extract of Viticis Fructus (Verbenaceae, WEVF) on experimental allergic reactions and inflammation. WEVF was anally administered to mice for high and fast absorption. WEVF inhibited compound 48/80-induced systemic anaphylaxis and immunoglobulin E (IgE)-mediated local allergic reaction. Histamine releasing from mast cells was reduced by WEVF, which was mediated by modulation of intracellular calcium. In addition, WEVF decreased the gene expression of pro-inflammatory cytokines in phorbol 12-myristate 13-acetate plus calcium ionophore A23187 (PMACI)-stimulated HMC-1 cells. These findings demonstrate that the WEVF possesses antiallergic and anti-inflammatory activities, which may be mediated by reducing the release of mediators such as histamine from mast cells and weakening the inflammatory action of these mediators.

• Natural Product Sciences
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• 제26권4호
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• pp.295-302
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• 2020

Cudrania tricuspidata (C. tricuspidata) is a deciduous tree found in Japan, China and Korea. The root, stems, bark and fruit of C. tricuspidata has been used as traditional herbal remedies such as eczema, mumps, acute arthritis and tuberculosis. In this study, we investigated the potential efficacies of this natural compound by focusing on the inhibitory effect of cudraxanthone L (CXL) isolated from the roots of C. tricuspidata on human platelet aggregation. Our study focused on the action of CXL on collagen-stimulated human platelet aggregation, inhibition of platelet signaling molecules such as fibrinogen binding, intracellular calcium mobilization, fibronectin adhesion, dense granule secretion, and thromboxane A2 secretion. In addition, we investigated the inhibitory effect of CXL on thrombin-induced clot retraction. Our results showed that CXL inhibited collagen-induced human platelet aggregation, intracellular calcium mobilization, fibrinogen binding, fibronectin adhesion and clot retraction without cytotoxicity. Therefore, we confirmed that CXL has inhibitory effects on human platelet activities and has potential value as a natural substance for preventing thrombosis.

• Journal of Chest Surgery
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• 제25권4호
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• pp.364-382
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• 1992

In order to investigate the effect of intracellular cyclic GMP on the calcium channel, whole cell patch clamp technique with internal perfusion method was used in the single ventricular myocytes of the rabbit. Cyclic GMP, cGMP analogues, cAMP, isopernaline and forskolin were perfused into cells and their effects on the calcium current were analysed by applying depolarizing step pulse of 10 mV in amplitude for 200 msec from holding potential of -40 mV. Calcium currents usually activated from -30 mV and then reached a peak at +10 mV. Amplitude of the calcium current was standardized with membrane capacitance, 50 pF. Peak amplitude at +10 mV in control was -0.15 nA/50pF. When 100 mM cAMP was applied from the pipette, peak amplitude of calcium current increased to -0.32 nA and addition of 1 mM isoprenaline further increased its amplitude. In the presence of cGMP it alone also produced an increase of the calcium current to -0.52 nA/50pF and addition of isoprenaline or forskolin increased its magnitude to -[0.55~0.95] nA/50pF. Simultaneous application of cGMP and cAMP increased the calcium current to -0.67 nA/50pF. Among the cGMP analogues, 8-Br-cGMP was the most potent stimulant for the calcium current activation. From the above results it could be concluded tlat cGMP increases the calcium current not through cAMP dependent protein kinase nor cAMP dependent phosphodiesterase pathway, but through independent phosphorylation pathway, possibly cGMP dependent protein kinase pathway.

• Toxicological Research
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• 제16권1호
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• pp.63-66
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• 2000

The effects of acute ethanol on the high K+ induced $Ca^{2+}}$ signals were examined from primary cultures of cerebellar granule neurons. $Ca^{2+}}$ signals were measured with Calcium Green-1 based microscopic video imaging. Because $Ca^{2+}}$ signal was low in most of granule neurons without stimuli, high KCI was used for depolarization. In most case, acute exposure to ethanol reduced the peak amplitude of the $Ca^{2+}}$ signals, induced by high K+, even though low concentration of ethanol(2~10mM) was used and the effects lasted more than 30min. In was also possible to see differences of ethanol inhibition, i.e. the temporal pattern of $Ca^{2+}}$ signal reductions and the strength of inhibition of $Ca^{2+}}$ signals in cerebellar granule neurons. These results indicate that low concentration of ethanol has diverse actions on the $Ca^{2+}}$ signals in cerebellar granule neurons.

• 한국발생생물학회지:발생과생식
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• 제19권2호
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• pp.61-67
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• 2015

The immediate early gene c-fos has long been known as a molecular marker of neural activity. The neuron's activity is transformed into intracellular calcium influx through NMDA receptors and L-type voltage sensitive calcium channels. For the transcription of c-fos, neural activity should be strong enough to activate mitogen-activated protein kinase (MAPK) signaling pathway which shows low calcium sensitivity. Upon translation, the auto-inhibition by Fos protein regulates basal Fos expression. The pattern of external stimuli and the valence of the stimulus to the animal change Fos signal, thus the signal reflects learning and memory aspects. Understanding the features of multiple components regulating Fos signaling is necessary for the optimal generation and interpretation of Fos signal.