• Archives of Pharmacal Research
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• v.29 no.10
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• pp.834-839
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• 2006

Torilin was purified from Torilis japonica (Houtt.) DC., and its effects on a rapidly activating delayed rectifier $K^+$ channel (hKv1.5), cloned from human heart and stably expressed in Ltk cells, as well as the corresponding $K^+$ current (the ultrarapid delayed rectifier, $I_{KUR}$) were assessed in human atrial myocytes. Using the whole cell configuration of the patch-clamp technique, torilin was found to inhibit the hKv1.5 current in time and voltage-dependent manners, with an $IC_50$ value of $2.51{\pm}0.34\;{\mu}M$ at +60 mV. Torilin accelerated the inactivation kinetics of the hKv1.5 channel, and slowed the deactivation kinetics of the hKv1.5 current, resulting in a tail crossover phenomenon. Additionally, torilin inhibited the hKv1.5 current in a use dependent manner. These results strongly suggest that torilin is a type of open-channel blocker of the hKv1.5 channel.

• International Journal of Oral Biology
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• v.42 no.2
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• pp.55-61
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• 2017

Recent studies indicate that mitochondria are an important source of reactive oxygen species (ROS) in the spinal dorsal horn. In our previous study, application of malate, a mitochondrial electron transport complex I substrate, induced a membrane depolarization, which was inhibited by pretreatment with ROS scavengers. In the present study, we used patch clamp recording in the substantia geletinosa (SG) neurons of spinal slices, to investigate the cellular mechanism of mitochondrial ROS on neuronal excitability. DNQX (an AMPA receptor antagonist) and AP5 (an NMDA receptor antagonist) decreased the malate-induced depolarization. In an external calcium free solution and addition of tetrodotoxin (TTX) for blockade of synaptic transmission, the malate-induced depolarization remained unchanged. In the presence of DNQX, AP5 and AP3 (a group I metabotropic glutamate receptor (mGluR) antagonist), glutamate depolarized the membrane potential, which was suppressed by PBN. However, oligomycin (a mitochondrial ATP synthase inhibitor) or PPADS (a P2 receptor inhibitor) did not affect the substrates-induced depolarization. These results suggest that mitochondrial substrate-induced ROS in SG neuron directly acts on the postsynaptic neuron, therefore increasing the ion influx via glutamate receptors.

• The Korean Journal of Physiology and Pharmacology
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• v.22 no.3
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• pp.249-255
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• 2018

Echinacoside, an active compound in the herb Herba Cistanche, has been reported to inhibit glutamate release. In this study, we investigated the effects of echinacoside on spontaneous excitatory synaptic transmission changes induced by 4-aminopyridine (4-AP), by using the in vitro rat hippocampal slice technique and whole-cell patch clamp recordings from CA3 pyramidal neurons. Perfusion with echinacoside significantly suppressed the 4-AP-induced epileptiform activity in a concentration-dependent manner. Echinacoside reduced 4-AP-induced increase in frequency of spontaneous excitatory postsynaptic currents (sEPSCs) but it did not affect the amplitude of sEPSCs or glutamate-activated currents, implicating a presynaptic mechanism of action. Echinacoside also potently blocked sustained repetitive firing, which is a basic mechanism of antiepileptic drugs. These results suggest that echinacoside exerts an antiepileptic effect on hippocampal CA3 pyramidal neurons by simultaneously decreasing glutamate release and blocking abnormal firing synchronization. Accordingly, our study provides experimental evidence that echinacoside may represent an effective pharmacological agent for treating epilepsy.

• Animal cells and systems
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• v.15 no.2
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• pp.95-106
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• 2011

The transient receptor potential vanilloid 4 (TRPV4) cation channel, a member of the TRP vanilloid subfamily, is expressed in a broad range of tissues. Nitric oxide (NO) as a gaseous signal mediator shows a variety of important biological effects. In many instances, NO has been shown to exhibit its activities via a protein S-nitrosylation mechanism in order to regulate its protein functions. With functional assays via site-directed mutagenesis, we demonstrate herein that NO induces the S-nitrosylation of TRPV4 $Ca^{2+}$ channel on the $Cys^{853}$ residue, and the S-nitrosylation of $Cys^{853}$ reduced its channel sensitivity to 4-${\alpha}$ phorbol 12,13-didecanoate and the interaction between TRPV4 and calmodulin. A patch clamp experiment and $Ca^{2+}$ image analysis show that the S-nitrosylation of $Cys^{853}$ modulates the TRPV4 channel as an inhibitor. Thus, our data suggest a novel regulatory mechanism of TRPV4 via NO-mediated S-nitrosylation on its $Cys^{853}$ residue.

• The Korean Journal of Physiology and Pharmacology
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• v.25 no.3
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• pp.251-258
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• 2021

Flos magnoliae (FM), the dry flower buds of Magnolia officinalis or its related species, is a traditional herbal medicine commonly used in Asia for symptomatic relief of and treating allergic rhinitis, headache, and sinusitis. Although several studies have reported the effects of FM on store-operated calcium entry (SOCE) via the ORAI1 channel, which is essential during intracellular calcium signaling cascade generation for T cell activation and mast cell degranulation, the effects of its isolated constituents on SOCE remain unidentified. Therefore, we investigated which of the five major constituents of 30% ethanoic FM (vanillic acid, tiliroside, eudesmin, magnolin, and fargesin) inhibit SOCE and their physiological effects on immune cells. The conventional whole-cell patch clamp results showed that fargesin, magnolin, and eudesmin significantly inhibited SOCE and thus human primary CD4+ T lymphocyte proliferation, as well as allergen-induced histamine release in mast cells. Among them, fargesin demonstrated the most potent inhibitory effects not only on ORAI1 (IC50 = 12.46 ± 1.300 μM) but also on T-cell proliferation (by 87.74% ± 1.835%) and mast cell degranulation (by 20.11% ± 5.366%) at 100 μM. Our findings suggest that fargesin can be a promising candidate for the development of therapeutic drugs to treat allergic diseases.

• The Korean Journal of Physiology and Pharmacology
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• v.25 no.3
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• pp.227-237
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• 2021

Carbon monoxide (CO) is a cardioprotectant and potential cardiovascular therapeutic agent. Human cardiac fibroblasts (HCFs) are important determinants of myocardial structure and function. Large-conductance Ca2+-activated K+ (BK) channel is a potential therapeutic target for cardiovascular disease. We investigated whether CO modulates BK channels and the signaling pathways in HCFs using whole-cell mode patch-clamp recordings. CO-releasing molecules (CORMs; CORM-2 and CORM-3) significantly increased the amplitudes of BK currents (IBK). The CO-induced stimulating effects on IBK were blocked by pre-treatment with specific nitric oxide synthase (NOS) blockers (L-NG-monomethyl arginine citrate and L-NG-nitroarginine methyl ester). 8-bromo-cyclic GMP increased IBK. KT5823 (inhibits PKG) or ODQ (inhibits soluble guanylate cyclase) blocked the CO-stimulating effect on IBK. Moreover, 8-bromo-cyclic AMP also increased IBK, and pre-treatment with KT5720 (inhibits PKA) or SQ22536 (inhibits adenylate cyclase) blocked the CO effect. Pre-treatment with N-ethylmaleimide (a thiol-alkylating reagent) also blocked the CO effect on IBK, and DL-dithiothreitol (a reducing agent) reversed the CO effect. These data suggest that CO activates IBK through NO via the NOS and through the PKG, PKA, and S-nitrosylation pathways.

• The Korean Journal of Physiology and Pharmacology
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• v.27 no.1
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• pp.95-103
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• 2023

Rosiglitazone is a thiazolidinedione-class antidiabetic drug that reduces blood glucose and glycated hemoglobin levels. We here investigated the interaction of rosiglitazone with Kv3.1 expressed in Chinese hamster ovary cells using the wholecell patch-clamp technique. Rosiglitazone rapidly and reversibly inhibited Kv3.1 currents in a concentration-dependent manner (IC₅₀ = 29.8 µM) and accelerated the decay of Kv3.1 currents without modifying the activation kinetics. The rosiglitazonemediated inhibition of Kv3.1 channels increased steeply in a sigmoidal pattern over the voltage range of -20 to +30 mV, whereas it was voltage-independent in the voltage range above +30 mV, where the channels were fully activated. The deactivation of Kv3.1 current, measured along with tail currents, was also slowed by the drug. In addition, the steady-state inactivation curve of Kv3.1 by rosiglitazone shifts to a negative potential without significant change in the slope value. All the results with the use dependence of the rosiglitazone-mediated blockade suggest that rosiglitazone acts on Kv3.1 channels as an open channel blocker.

• Journal of Yeungnam Medical Science
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• v.19 no.2
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• pp.99-106
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• 2002

Background: Simple ventricular septal defect(VSD) is the most common congenital heart disease. Although closure of VSD is currently associated with a relatively low risk, experience with younger and smaller infants has been variably less satisfactory. We assessed the results of surgical closure of VSD in infant. Materials and Methods: Between 1996 and 2000, 45 non-restrictive VSD patients underwent patch repair and retrospective analysis was done. Patients were divided into two groups based on weight: group I infants weighed 5kg or less(n=16), and group II infants weighed more than 5kg(n=29). Both groups had similar variation in sex, VSD location, aortic cross clamp time and total bypass time. But combined diseases (ASD, PDA, MR) were more in group I. We closed VSD with patch and used simple continuous suture method in all patients. Results: There were no operative mortality, no reoperation for hemodynamically significant residual shunt and no surgically induced complete heart block. As a complication, pneumonia(group I: 2 cases, group II: 2 cases), transient seizure(group II: 2), wound infection(group I: 1, group II: 1), urinary tract infection(group I: 1) and chylopericardium(group I: 1) developed, and there was no significant difference between two groups(p>0.05). Conclusion: Early primary closure with simple continuous suture method was applicable in all patients with non-restrictive VSD without any serious complications.

• Progress in Medical Physics
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• v.14 no.1
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• pp.54-67
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• 2003

The voltage-dependence of N-type calcium current inactivation is U-shaped with the degree of inactivation roughly mirroring inward current. This voltage-dependence has been reported to result from a purely voltage-dependent mechanism. However, $Ca^{2＋}$-dependent inactivation of N-channels has also been reported. We have investigated the role of $Ca^{2＋}$ in N-channel inactivation by comparing the effects of $Ba^{2＋}$and $Ca^{2＋}$ on whole-cell N-current in rat superior cervical ganglion neurons. For individual cells in-activation was always larger in $Ca^{2＋}$ than in $Ba^{2＋}$ even when internal EGTA (11 mM) was replaced with BAPTA (20 mM). The inactivation vs. voltage relationship was U-shaped in both divalent cations. The enhancement of inactivation by $Ca^{2＋}$ was inversely related with the magnitude of inactivation in $Ba^{2＋}$ as if the mechanisms of inactivation were the same in both $Ba^{2＋}$ and $Ca^{2＋}$. In support of this idea we could separate fast ( ${\gamma}$ ～150 ms) and slow ( ${\gamma}$ ～ 2500 ms) components of inactivation in both $Ba^{2＋}$and $Ca^{2＋}$ using 5 sec voltage steps. Differential effects were observed on each component with $Ca^{2＋}$ enhancing the magnitude of the fast component and the speed of the slow component. The larger amplitude of fast component indicates that the more channels inactivate via this pathway with $Ca^{2＋}$ than with $Ba^{2＋}$, but the stable time constants support the idea the fast inactivation mechanism is identical in $Ba^{2＋}$and $Ca^{2＋}$. The results do not support a $Ca^{2＋}$-dependent mechanism for fast inactivation. However, the $Ca^{2＋}$-induced acceleration of the slowly inactivating component could result from a $Ca^{2＋}$-dependent process.

• The Journal of Korean Medicine
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• v.19 no.2
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• pp.450-467
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• 1998

To research the characteristics of ion currents induced by inhibitory and excitatory herbs of oriental medicine, we used nystatin-perforated patch clamp technique under voltage clamp condition in periaqueductal gray neuron dissociated from Sprauge-Dawley rat, 10-15 days old. The results are as follows. 1. Ion current induced by $10mg/m{\ell}$ of Bupleuri Radix was inhibited $59.50{\pm}4.29%$ by $10^{-4}M$ bicuculline(p>0.01) but inhibition of $10.75{\pm}4.77%$ by $10^{-4}M$ tubocurarine and $4.75{\pm}4.23%$ by $10^{-4}M$ verapamil had no statistical significance(p>0.05). So ion current induced by Bupleuri Radix revealed only GABA induced $Cl^-$ current, not acetylcholine and $Ca^{2+}$ current. 2. Ion current induced by $20mg/m{\ell}$ of Coptidis Rhizoma was inhibited $47.20{\pm}7.88%$ by $10^{-4}M$ bicuculline(p<0.01) but $3.20{\pm}2.33%$ inhibition by $10^{-4}M$ tubocurarine and $1.00{\pm}1.00%$ inhibition by $10^{-4}M$ verapamil had no significance(p>0.05). So ion current induced by Coptidis Rhizoma revealed only GABA induced $Cl^-$ current, not acetylcholine and $Ca^{2+}$ current. 3. Ion current induced by $20mg/m{\ell}$ of Ecliptae Herba was inhibited $55.00{\pm}4.92%$ by $10^{-4}M$ bicuculline (p<0.01), and also inhibited $15.00{\pm}4.26%$ by $10^{-4}M$ tubocurarine(p<0.05), but inhibition of $6.00{\pm}3.03%$ by $10^{-4}M$ verapamil had no significance(p>0.05). So ion current induced by Ecliptae Herba showed GABA activated $Cl^-$ current and acetylcholine activated cation current, not $Ca^{2+}$ current 4. Ion current induced by $5mg/m{\ell}$ of Liriopis Tuber was inhibited $15.20{\pm}4.57%$ by $10^{-4}M$ bicuculline<0.05) and also inhibited $14.00{\pm}3.00%$ by $10^{-4}M$ tubocurarine(p<0.05), but inhibition of $5.20{\pm}4.80%$ by $10^{-4}M$ verapamil had no significance(p>0.05). So ion current induced by Liriopis Tuber showed GABA. activated $Cl^-$ current and acetylcholine activated cation current, not $Ca^{2+}$ current. 5. Ion current induced by $5mg/m{\ell}$ of Aconiti Tuber was inhibited $97.00{\pm}1.34%$ by $10^{-4}M$ bicuculline(p<0.01), $80.00{\pm}9.83%$ by $10^{-4}M$ tubocurarine(p<0.01), and $24.00{\pm}6.18%$ by $10^{-4}M$ verapamil(p<0.05). So ion current induced by Aconiti Tuber revealed GABA activated $Cl^-$ current and acetylcholine activated cation current and $Ca^{2+}$ current. 6. Ion current induced by $10mg/m{\ell}$ of Zingiberis Rhizoma was inhibited $33.00{\pm}7.43%$ by $10^{-4}$ bicuculline(p<0.05), $10.20{\pm}1.83%$ by $10-^{-4}M$ tubocurarine(p<0.01), and $14.00{\pm}2.16%$ by $10^{-4}M$ verapamil(p<0.01) So ion current induced by Zingiberis Rhizoma revealed GABA activated $Cl^-$ current and acetylcholine activated cation outtent and $Ca^{2+}$ current. 7. Ion current induced by $10mg/m{\ell}$ of Boshniakiae Herba was inhibited $65.00{\pm}13.75%$ by $10^{-4}M$ bicuculline(p<0.05), $38.00{\pm}9.24%$ by $10^{-4}M$ tubocurarine(p<0.05), and $33.25{\pm}7.42%$ by $10^{-4}M$ verapamiHp<0.05). So ion current induced by Bpshniakiae Herba revealed GABA activated $Cl^-$ current and acetylcholine activated cation current and $Ca^{2+}$ current. These results suggest that a point of difference between inhibitory and excitatory herbs is existence of$Ca^{2+}$ current.