• 약학회지
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• 제43권5호
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• pp.659-664
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• 1999

To investigate the difference of contractile mechanism between KCI and phenylephrine-induced contraction, we observed effects of $Ca^{2+}$ antagonists and protein kinase inhibitors on aorta contraction of rats. Verapamil dose-dependently inhibited the contraction induced by KCI and phenylephrine, the inhibitory effect of verapamil was more potent in KCI-induced contraction than phenylephrine-induced contraction. Econazole and TMB-8 significantly inhibited CKI-induced contraction but did not inhibit phenylephrine-induced contraction. Staurosporine dose-dependently inhibited both KCI and phenylephrine-induced contraction. Genistein and calmodulin antagonists (W-7 and trifluoperazine) also inhibited both contraction in a dose dependent manner. However, the inhibitory effects of genistein and calmodulin antagonists were more potent in phenylephrine-induced contraction than KCI-induced contraction. These results suggest that involvements of $Ca^{2+}$ channel and protein kinase in rat aorta contraction were dependent on agonist causing aorta smooth muscle contraction.

• Natural Product Sciences
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• 제12권4호
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• pp.217-220
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• 2006

The effects of $(1R,9S)-\beta-hydrastine$ hydrochloride (BHSH) on $Ca^{2+}$ transport in rat pheochromocytoma PC12 cells were investigated. In the presence of external $Ca^{2+}$, BHSH at $100{\mu}M$ inhibited $K^+$ (56mM)-induced dopamine release, and $K^+-induced$ $Ca^{2+}$ influx and a sustained rise of $[Ca^{2+}]_i$. In addition, BHSH at 100 f.!M reduced the sustained rise of $[Ca^{2+}]_i$ elicited by 20 mM caffeine, but not by $1{\mu}M$ thapsigargin, in presence of external $Ca^{2+}$. These results suggest that BHSH inhibited $K^+-induced$ dopamine release and $[Ca^{2+}]_i$ influx, and store-operated $Ca^{2+}$ channels activated by caffeine, but not by thapsigargin, in PC12 cells.

• Molecules and Cells
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• 제37권11호
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• pp.804-811
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• 2014

The protease-activated receptor (PAR)-2 is highly expressed in endothelial cells and vascular smooth muscle cells. It plays a crucial role in regulating blood pressure via the modulation of peripheral vascular tone. Although several mechanisms have been suggested to explain PAR-2-induced hypotension, the precise mechanism remains to be elucidated. To investigate this possibility, we investigated the effects of PAR-2 activation on N-type $Ca^{2+}$ currents ($I_{Ca-N}$) in isolated neurons of the celiac ganglion (CG), which is involved in the sympathetic regulation of mesenteric artery vascular tone. PAR-2 agonists irreversibly diminished voltage-gated $Ca^{2+}$ currents ($I_{Ca}$), measured using the patch-clamp method, in rat CG neurons, whereas thrombin had little effect on $I_{Ca}$. This PAR-2-induced inhibition was almost completely prevented by ${\omega}$-CgTx, a potent N-type $Ca^{2+}$ channel blocker, suggesting the involvement of N-type $Ca^{2+}$ channels in PAR-2-induced inhibition. In addition, PAR-2 agonists inhibited $I_{Ca-N}$ in a voltage-independent manner in rat CG neurons. Moreover, PAR-2 agonists reduced action potential (AP) firing frequency as measured using the current-clamp method in rat CG neurons. This inhibition of AP firing induced by PAR-2 agonists was almost completely prevented by ${\omega}$-CgTx, indicating that PAR-2 activation may regulate the membrane excitability of peripheral sympathetic neurons through modulation of N-type $Ca^{2+}$ channels. In conclusion, the present findings demonstrate that the activation of PAR-2 suppresses peripheral sympathetic outflow by modulating N-type $Ca^{2+}$ channel activity, which appears to be involved in PAR-2-induced hypotension, in peripheral sympathetic nerve terminals.

• 약학회지
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• 제51권1호
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• pp.63-67
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• 2007

To investigate the relation between extracellular Ca$^{2+}$ and histamine release, we observed agonist-induced histamine release from RBL 2H3 mast cells in the presence or absence of extracellular Ca$^{2+}$ concentration. Histamine release induced by melittin and thapsigargin were greater in the presence of extracellular Ca$^{2+}$ than in the absence of extracellular Ca$^{2+}$. Econazole-induced histamine release had nothing to do with extracellular Ca$^{2+}$, whereas arachidonic acid-induced histamine release increased in the absence of extracellular Ca$^{2+}$. Calmodulin antagonists did not affect melittin-induced histamine release but they may potentiate arachidonic acid-induced histamine release. These data suggest that arachidonic acid-induced histamine release may be mediated via Ca$^{2+}$-independent pathway and may be potentiated by the block of Ca$^{2+}$-dependent pathway.

• 한국환경과학회:학술대회논문집
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• 한국환경과학회 2002년도 봄 학술발표대회 발표논문집
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• pp.471-474
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• 2002

SA treatment significantly increased thermotolerance In cucumber seedlings. Pretreatment of seeds with $CaCl_2$ solution enhanced the SA- induced thermotolerance. On the contrary, pretreatment with the $Ca^{2+}$ chelator EGTA lowered this SA-induced thermotolerance. In addition, pretreatment with $Ca^{2+}$ channel blocker verapamil also weakened the SA-induced thermotolerance. However, the calmodulin antagonist chlorpromazine(CPZ) had little effect on the SA-induced thermotolerance. Measurement of activity of the antioxidant enzyme APX and the level of lipid peroxidation (in term of MDA) indicated that heat stress induced an oxidative stress in cucumber seedlings. SA treatment induced higher activities of APX and a lower level of lipid peroxidation. $Ca^{2+}$ pretreatment further enhanced the SA-induced increase in APX activity and lowered the heat stress-induced lipid peroxidation, but EGTA pretreatment had a contrary effect. These results suggest that $Ca^{2+}$ and calmodulin may be involved In the acquisition of the SA-induced thermotolerance; antioxidant enzyme system take part in the final generation of the SA-induced thermotolerance.

• The Korean Journal of Physiology and Pharmacology
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• 제3권2호
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• pp.199-205
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• 1999

Vibrio vulnificus cytolysin caused platelet cytolysis and increased intracellular calcium concentration $([Ca^{2+}]_i)$ of rat platelets in a concentration-dependent manner. In the presence of V. vulnificus cytolysin (3 HU/ml), lactate dehydrogenase (LDH) activity was increased from $1.3{\pm}0.4%$ of control to $64.3{\pm}3.4%$ in platelet suspension buffer. In $Ca^{2+}-free$ platelet suspension buffer, however, V. vulnificus cytolysin did not induce $[Ca^{2+}]_i$ increase and LDH release. Addition of EGTA (2 mM) to suspension buffer after the initial $Ca^{2+}$ influx reversed $[Ca^{2+}]_i$ to the control level. However, a $Ca^{2+}$ channel blocker verapamil $(20\;{\mu}M)$ or mefenamic acid $(20\;{\mu}M)$ did not inhibit V. vulnificus cytolysin-induced $[Ca^{2+}]_i$ increase and LDH release. Divalent cations such as $Co^{2+},\;Cd^{2+}\;or\;Mn^{2+}$ (2 mM each) also did not alter V. vulnificus cytolysin-induced $[Ca^{2+}]_i$ increase and LDH release. V. vulnificus cytolysin (3 HU/ml)-induced calcium influx was completely blocked by lanthanum (2 mM). Lanthanum (2 mM) also completely blocked V. vulnificus cytolysin (3 HU/ml)-induced LDH release. Osmotic protectants such as, raffinose, sucrose or PEG600 (50 mM each) did not inhibit the lytic activity of V. vulnificus cytolysin. In conclusion, lanthanum sensitive $Ca^{2+}$ influx plays a significant role in Vibrio vulnificus cytolysin-induced platelet cytolysis and thrombocytopenia in V. vulnificus infection.

• The Korean Journal of Physiology
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• 제29권2호
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• pp.225-232
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• 1995

The present study was conducted to evaluate the effect of phorbol 12,13-dibutyrate (PDBu) on the contraction of rabbit jugular vein in vitro. PDBu concentrations of greater than 10 nM induced a periodic contraction which was composed of rapid contraction, plateau and slow relaxation. The frequency of periodic contraction increased as PDBu concentration increased. The PDBu-induced contraction was inhibited by staurosporine (100 nM), it was not changed by tetrodotoxin $(1\;{\mu}M).$ In $Ca^{2+}$-free medium, PDBu induced a sustaining contraction, but not periodic contraction. Addition of $Ca^{2+}$ to medium evoked periodic contraction which was inhibited by nifedipine, PDBu concentrations of greater than $0.1\;{\mu}M$ increased ^{45}Ca^{2+}$ uptake without changing $^{45}Ca^{2+}$ efflux. Charybdotoxin and apamin, $Ca^{2+}$-activated K^{+}$ channel blockers, did not affect the PDBu-induced periodic contraction, whereas tetraethylammonium (TEA) abolished the periodicity. Pinacidil $(10\;{\mu}M).$, a potassium channel activator, blocked PDBu induced periodic contraction, which was recovered by glybenclamide $(10\;{\mu}M).$. In high potassium solution, PDBu did not produce the periodic contraction. These results suggest that the PDBu-induced periodicity of contraction is modulated by voltage dependent $Ca^{2+}$ channel and ATP-sensitive $K^{+}$ channel.

• The Korean Journal of Physiology and Pharmacology
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• 제20권1호
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• pp.15-23
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• 2016

Acetaminophen (APAP) overdose is one of the most common causes of acute liver failure. The study aimed to investigate the protective effect of carnosic acid (CA) on APAP-induced acute hepatotoxicity and its underlying mechanism in mice. To induce hepatotoxicity, APAP solution (400 mg/kg) was administered into mice by intraperitoneal injection. Histological analysis revealed that CA treatment significantly ameliorated APAP-induced hepatic necrosis. The levels of both alanine aminotransferase (ALT) and aspartate transaminase (AST) in serum were reduced by CA treatment. Moreover, CA treatment significantly inhibited APAP-induced hepatocytes necrosis and lactate dehydrogenase (LDH) releasing. Western blot analysis showed that CA abrogated APAP-induced cleaved caspase-3, Bax and phosphorylated JNK protein expression. Further results showed that CA treatment markedly inhibited APAP-induced pro-inflammatory cytokines TNF-${\alpha}$, IL-$1{\beta}$, IL-6 and MCP-1 mRNA expression and the levels of phosphorylated $I{\kappa}B{\alpha}$ and p65 protein in the liver. In addition, CA treatment reduced APAP- induced hepatic malondialdehyde (MDA) contents and reactive oxygen species (ROS) accumulation. Conversely, hepatic glutathione (GSH) level was increased by administration of CA in APAP-treated mice. Mechanistically, CA facilitated Nrf2 translocation into nuclear through blocking the interaction between Nrf2 and Keap1, which, in turn, upregulated anti-oxidant genes mRNA expression. Taken together, our results indicate that CA facilitates Nrf2 nuclear translocation, causing induction of Nrf2-dependent genes, which contributes to protection from acetaminophen hepatotoxicity.

• The Korean Journal of Physiology
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• 제18권2호
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• pp.125-137
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• 1984

Vanadate의 수축에 이용되는 $Ca^{2+}$의 동원 경로와 Na-Pump억제가 vanadate의 수축에 어떤 영향을 미치는 지를 밝히기 위해 본 실험을 시행하여 다음과 같은 곁과를 얻었다. 1) 흰쥐의 자궁근에서는 vanadate는 수축을 일으켜 $5{\times}10^{-4}M$에서 최대수축을 나타내었으며 사람의 자궁근이 흰쥐의 자중근에 비해 vanadate에 더 민감한 반응을 보였다. 2) Vanadate에 의한 수축은 $Ca^{2+}$제거에 의해 완전히 억제되지 않았고 사람의 자궁근이 외부 $Ca^{2+}$의 농도변화에 더 민감한 반응을 보였다. 3) Vanadate에 의한 수축은 verapamil농도를 증가시킴에 따 억제되었으며 100k에 극한 수축을 완전 억제시키는$3{\times}10^{-5}M$ verapamil 존재하에서도 최대수의 40%정도가 남아있었고, 이 크기는 $Ca^{2+}$없는 용액에서의 수축의 크기와 유사하였다. 4) Na-pump억제시 vanadate의 수축은 증가하였고 이 현상은 $3{\times}10^{-5}M$ verapamil 존재하에서도 나타났다. 5) $Ca^{2+}$없는 ouabain용액에서 전처치후에 vanadate에 의한 수축은 증가하지 않았으나 외부내 $Ca^{2+}$을 부가할 나타나는 반음은 대조군에 비해 현저히 증가하였다. 6) Verapamil 존재시 vanadate에 의한 $Ca^{45}$유입은 완전히 억제되었으나 ouabain으로 처리한 후는 verapamil 존재하에서도 vanadate가 현저히 $Ca^{45}$유입을 일으켰다. 7) Ouabain이나 K 없는 용액으로 치리시간이 증가함에 따라 vanadate에 의한 수축의 증가정도는 더욱 더 현저하였다. 8) Ouabain 전처치시 증가된 vanadate에 의한 수축은 $10^{-4}M$ papaverine에 의해 현저히 억제되었다. 9) Acetylcholine에 의한 수축은 verapamil 존재하에서도 Na-pump억제 시간이 증가함에 따라 증가하였다. 이상의 결과로 볼 때 vanadate에 대해 사람의 자궁근이 흰쥐의 자궁근에 비해 더 민감한 반응을 보이고 vanadate에 의한 수축에는 외부와 내부 $Ca^{2+}$이 모두 이용되며 Na-pump 억제시 여러가지 근수축물질이 verapamil에 의해 억제되지 않는 $Ca^{2+}$유입을 일으키며 이 유입경로의 성질은 확실히 알 수 없으나 Papaverine에 의해 억제되며 막전위의 변화와 관련이 있는 것으로 생각된다.

• Journal of Korean Biological Nursing Science
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• 제1권1호
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• pp.25-41
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• 1999

Physiological activity of osteoblast including bone formation is known to be closely related to the increase of intracellular $Ca^{2+}$ activity($[Ca^{2+}]_i$) in osteoblast. $Ca^{2+}$ is an important intracellular messenger in diverse cellular functions, and regulation of its level is mediated by the transmembrane $Ca^{2+}$ movement via $Ca^{2+}$ channels, $Na^+-Ca^{2+}$ exchange, and by intracellular $Ca^{2+}$ movement through the intracellular stores. The purpose of this study is to investigate how the intracellular $Ca^{2+}$ is regulated in osteoblast-like cells(OLCs) by measuring $Ca^{2+}$ activity with cell imaging technique. OLCs were isolated from femur and tibia of neonatal rats, and cultured for 7 days. Cultured OLCs were loaded with a $Ca^{2+}$-sensitive fluorescent dye, Fura-2, and fluorescence images were monitored with a cooled CCD camera. The images were processed and analyzed with an image analyzing software. The results were as follows. (1) $[Ca^{2+}]_i$ of OLC decreased as the $Ca^{2+}$ concentration in the superfusing Tyrode solution was lowered. When $Na^+$ concentration in the superfusing solution was decreased, $[Ca^{2+}]_i$ increased.. These suggest that $Ca^{2+}$ flux occurs via the $Na^+-Ca^{2+}$ exchange mechanism. (2) When $Na^+$ in the superfusing solution was removed. a transient $Ca^{2+}$, increase($Ca^{2+}$ spike) was occasionally observed. However, $Ca^{2+}$ spike was not observed after adding 1 ${\mu}M$ thapsigargin. This implies that the generation of $Ca^{2+}$ spike is mediated by the release of $Ca^{2+}$ from endoplasmic reticulum(ER). (3) As the $Ca^{2+}$ concentration in the superfusing solution was raised, the frequency of 0mM $Na^+$-induced $Ca^{2+}$ spike increased, suggesting that $Ca^{2+}$-induced $Ca^{2+}$ release(CICR) mechanism exists. (4) After $[Ca^{2+}]_i$ was decreased with the superfusion of $Ca^{2+}$-free solution containing thapsigargin, the recovery of $[Ca^{2+}]_i$ with reperfusion of 2.5mM $Ca^{2+}$ solution transiently exceeded the control level, suggesting that the depletion of $Ca^{2+}$ in ER induces $Ca^{2+}$ influx from extracellular medium via store-operated $Ca^{2+}$ influx(SOCI) mechanism. (5) $[Ca^{2+}]_i$ was not affected by the superfusion of 25mM $K^+$ Tyrode solution. These results suggest that intracellular $Ca^{2+}$ activity in osteoblast is regulated by transmembrane $Ca^{2+}$ flux via $Na^+-Ca^{2+}$ exchange, $Ca^{2+}$ release from the internal store (ER) via $Ca^{2+}$-induced $Ca^{2+}$ release, and store-operated $Ca^{2+}$ influx across the cell membrane.