• Journal of Ginseng Research
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• v.44 no.3
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• pp.490-495
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• 2020

Background: Ginsenoside Rk1, a saponin component isolated from heat-processed Panax ginseng Meyer, has been implicated in the regulation of antitumor and anti-inflammatory activities. Although our previous studies have demonstrated that ginsenoside Rg3 significantly attenuated the activation of NMDA receptors (NMDARs) in hippocampal neurons, the effects of ginsenosides Rg5 and Rk1, which are derived from heat-mediated dehydration of ginsenoside Rg3, on neuronal NMDARs have not yet been elucidated. Methods: We examined the regulation of NMDARs by ginsenosides Rg5 and Rk1 in cultured rat hippocampal neurons using fura-2-based calcium imaging and whole-cell patch-clamp recordings. Results: The results from our investigation showed that ginsenosides Rg3 and Rg5 inhibited NMDARs with similar potencies. However, ginsenoside Rk1 inhibited NMDARs most effectively among the five compounds (Rg3, Rg5, Rk1, Rg5/Rk1 mixture, and protopanaxadiol) tested in cultured hippocampal neurons. Its inhibition is independent of the NMDA- and glycine-binding sites, and its action seems to involve in an interaction with the polyamine-binding site of the NMDAR channel complex. Conclusion: Taken together, our results suggest that ginsenoside Rk1 might be a novel component contributable to the development of ginseng-based therapeutic treatments for neurodegenerative diseases.

• Biomolecules & Therapeutics
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• v.28 no.4
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• pp.370-379
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• 2020

Econazole, a potent broad-spectrum antifungal agent and a Ca²⁺ channel antagonist, induces cytotoxicity in leukemia cells and is used for the treatment of skin infections. However, little is known about its cytotoxic effects on solid tumor cells. Here, we investigated the molecular mechanism underlying econazole-induced toxicity in vitro and evaluated its regulatory effect on the metastasis of gastric cancer cells. Using the gastric cancer cell lines AGS and SNU1 expressing wild-type p53 we demonstrated that econazole could significantly reduce cell viability and colony-forming (tumorigenesis) ability. Econazole induced G0/G1 phase arrest, promoted apoptosis, and effectively blocked proliferation- and survival-related signal transduction pathways in gastric cancer cells. In addition, econazole inhibited the secretion of matrix metalloproteinase- 2 (MMP-2) and MMP-9, which degrade the extracellular matrix and basement membrane. Econazole also effectively inhibited the metastasis of gastric cancer cells, as confirmed from cell invasion and wound healing assays. The protein level of p53 was significantly elevated after econazole treatment of AGS and SNU1 cells. However, apoptosis was blocked in econazole-treated cells exposed to a p53-specific small-interfering RNA to eliminate p53 expression. These results provide evidence that econazole could be repurposed to induce gastric cancer cell death and inhibit cancer invasion.

• The Korean Journal of Physiology and Pharmacology
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• v.5 no.3
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• pp.231-241
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• 2001

The mammalian cortical collecting duct (CCD) plays a major role in regulating renal NaCl reabsorption, which is important in $Na^+$ and $Cl^-$ homeostasis. The M-1 cell line, derived from the mouse cortical collecting duct, has been used as a mammalian model of the study on the electrolytes transport in CCD. M-1 cells were grown on collagen-coated permeable support and short circuit current $(I_{sc})$ was measured. M-1 cells developed amiloride-sensitive current $5{\sim}7$ days after seeding. Apical and basolateral addition of ATP induced increase in $I_{sc}$ in M-1 cells, which was partly retained in $Na^+-free$ or $Cl^--free$ solution, indicating that ATP increased $Na^+$ absorption and $Cl^-$ secretion in M-1 cells. $Cl^-$ secretion was mediated by the activation of apical cystic fibrosis transmembrane regulator (CFTR) chloride channels and $Ca^{2+}-activated$ chloride channels, but $Na^+$ absorption was not mediated by activation of epithelal sodium channel (ENaC). ATP increased cAMP content in M-1 cells. The RT-PCR analysis demonstrated that M-1 cells express $P2Y_2,\;P2X_3\;and\;P2Y_4$ receptors. These results showed that ATP regulates $Na^+$ and $Cl^-$ transports via multiple P2 purinoceptors on the apical and basolateral membranes in M-1 cells.

• The Korean Journal of Physiology and Pharmacology
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• v.3 no.1
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• pp.101-117
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• 1999

Amyotrophic lateral sclerosis (ALS) is a degenerative neuromuscular disease of unknown etiology in which the upper and lower motor neurons are progressively destroyed. Recent evidences support the role of autoimmune mechanisms in the pathogenesis of ALS. This study investigated the effects of sera from ALS patients on neuromuscular transmission in phrenic nerve-hemidiaphragm preparations and on calcium currents of single isolated dorsal root ganglion (DRG) cells in mice. Mice were injected with either control sera from healthy adults or ALS sera from 18 patients with ALS of sporadic form, for three days. Miniature end plate potential (MEPP) and nerve-evoked end plate potential (EPP) were measured using intracellular recording technique and the quantal content was determined. Single isolated DRG cells were voltage-clamped with the whole-cell configuration and membrane currents were recorded. Sera from 14 of 18 ALS patients caused a significant increase in MEPP frequency in normal Ringer's solution $(4.62{\pm}0.14\;Hz)$ compared with the control $(2.18{\pm}0.15\;Hz).$ In a high $Mg^{2+}/low\;Ca^{2+}$ solution, sera from 13 of 18 ALS patients caused a significant increase in MEPP frequency, from $2.18{\pm}0.31$ Hz to $6.09{\pm}0.38$ Hz. Sera from 11 of 18 patients produced a significant increase of nerve-evoked EPP amplitude, from $0.92{\pm}0.05$ mV to $1.30{\pm}0.04$ mV, while the other seven ALS sera did not alter EPP amplitude. In the ALS group, EPP quantal content was also elevated by the sera of 14 patients (from $1.49{\pm}0.07$ to $2.35{\pm}0.07).$ MEPP frequency and amplitude in wobbler mouse were $4.03{\pm}0.53$ Hz and $1.37{\pm}0.18$ mV, respectively, which were significantly higher than those of wobbler controls (wobblers without the symptoms of wobbler). Sera from ALS patients significantly reduced HVA calcium currents of DRG cells to 42.7% at -10 mV. Furthermore, the inactivation curve shifted to more negative potentials with its half-inactivation potential changed by 6.98 mV. There were, however, significant changes neither in the reversal potential of $I_{Ca}$ nor in the I-V curve. From these results it was concluded that: 1) The serum factors of sporadic ALS patients increase neuromuscular transmission and can alter motor nerve terminal presynaptic function. This suggests that ALS serum factors may play an important role in the early stage of ALS, and 2) Calcium currents in DRG cells were reduced and rapidly inactivated by ALS sera, suggesting that in these cells, ALS serum factors may exert interaction with the calcium channel.

• Journal of Life Science
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• v.34 no.6
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• pp.426-433
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• 2024

The environmental stress that plants are most susceptible to is water stress. Abscisic acid (ABA) is a plant hormone synthesized by plants to counteract environmental stress. The role of stomata in plants is to allow the synthesis of sucrose by absorbing CO₂, which greatly affects photosynthetic activity. In addition, stomata are pathways for transpiration, which releases H₂O and help establish a water potential gradient that allows plant roots to continuously absorb water and inorganic substances from the soil. Plants have a mechanism to minimize water loss by closing their stomata when exposed to water-stressed environments. The most well-studied hypothesis concerning the mechanism of stomatal closure is the response to water stress. When a plant receives sufficient water, its stomata open during the day and close at night due to its circadian rhythm. In addition, stomatal closure occurs when the concentration of CO₂ in the intercellular space increases. However, the mechanism of stomatal closure due to circadian rhythm and increased CO₂ concentration in the intercellular space is not well understood. When plants undergo water stress, the increased concentration of ABA in the guard cell cytoplasm induces an increase in Ca²⁺ concentration, resulting in cytoplasmic depolarization. As a result, the outward K⁺-channel of the tonoplast and the slow-type anion channels SLAC1 and SLAH3 are activated, releasing K⁺, Cl^-, and malate^2-, causing the stomata to close. Therefore, in this paper, the mechanism of stomatal closure caused by water stress was investigated.

• Nuclear Engineering and Technology
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• v.18 no.3
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• pp.200-208
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• 1986

The loss of coolant accident based on a double-ended cold leg break is analyzed with the discharge coefficient (Ca) of 0.4. This analysis covers the whole transient period from the start of depressurization to the complete refilling of the core by using RELAP4/MOD6-EM and RELAP4/ MOD6-HOT CHANNEL for the system thermal-hydraulics and the fuel performance during the blowdown phase respectively, and RELAP4/MOD6-FLOOD and TOODEE2 during the reflood phase. A simple analytical method has been developed to account for the lower plenum filling by approximating steam-water countercurrent flows and superheated wall effects at the downcomer during the refill period. Based on the informations. at the time of EOB (end-of-bypass), the refill duration time and the initial reflooding temperature were estimated and compared with the results from the RELAP4/MOD6, resulting in a good agreement. In addition, some parametric studies on the EOB were performed. The form loss coefficient between upper head and upper downcomer was found to be sensitive to the occurrence of the spurious EOB. Appropriate form loss coefficients should be taken into account to avoid the flow oscillations at the downcomer. The analyses with the six and three volume core nodalizations, respectively, show much similar trends in the system thermal-hydraulic performance, but the former case is recommended to obtain good results.

• Korean Journal of Veterinary Research
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• v.34 no.1
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• pp.37-47
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• 1994

The effects of the novel compound GS-386 on the calcium current were investigated in rabbit atrial myocytes. The calcium current was recorded during various depolarizations of 200 ms duration from a holding potential of -40 mV using the whole cell patch clamp technique. The calcium current was activated from -30 mV, reached maximum amplitude at +10 mV and almost disappeared at +50 mV. Superfusion of GS-386 led to a reduction of the calcium current amplitude dose-dependently and $ED_{50}$ was $2.5{\times}10^{-7}M$. But the dependence of the calcium current on the membrane potential was not altered by GS-386. The inactivation of the calcium currents showed single exponential curves in both before and after application of GS-386. The inactivation time constants before and after application of GS-386 were almost the same(35 ms and 32.5 ms). The steady-state inactivation curve of the calcium current was not shifted by GS-386. The calcium currents both before and after application of GS-386 recovered completely in 1 sec and the recovery time constants were about 200 ms in both cases. From the above results it is concluded that the novel compound GS-386 has calcium antagonistic property decreasing the calcium current.

• The Korean Journal of Physiology and Pharmacology
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• v.24 no.6
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• pp.529-543
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• 2020

In contrast to ventricular myocytes, the structural and functional importance of atrial transverse tubules (T-tubules) is not fully understood. Therefore, we investigated the ultrastructure of T-tubules of living rat atrial myocytes in comparison with ventricular myocytes. Nanoscale cell surface imaging by scanning ion conductance microscopy (SICM) was accompanied by confocal imaging of intracellular T-tubule network, and the effect of removal of T-tubules on atrial excitation-contraction coupling (EC-coupling) was observed. By SICM imaging, we classified atrial cell surface into 4 subtypes. About 38% of atrial myocytes had smooth cell surface with no clear T-tubule openings and intracellular T-tubules (smooth-type). In 33% of cells, we found a novel membrane nanostructure running in the direction of cell length and named it 'longitudinal fissures' (LFs-type). Interestingly, T-tubule openings were often found inside the LFs. About 17% of atrial cells resembled ventricular myocytes, but they had smaller T-tubule openings and a lower Z-groove ratio than the ventricle (ventricular-type). The remaining 12% of cells showed a mixed structure of each subtype (mixed-type). The LFs-, ventricular-, and mixed-type had an appreciable amount of reticular form of intracellular T-tubules. Formamide-induced detubulation effectively removed atrial T-tubules, which was confirmed by both confocal images and decreased cell capacitance. However, the LFs remained intact after detubulation. Detubulation reduced action potential duration and L-type Ca²⁺ channel (LTCC) density, and prolonged relaxation time of the myocytes. Taken together, we observed heterogeneity of rat atrial T-tubules and membranous ultrastructure, and the alteration of atrial EC-coupling by disruption of T-tubules.

• Biomedical Science Letters
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• v.14 no.2
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• pp.69-76
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• 2008

${\gamma}$-aminobutyric acid (GABA) is the major inhibitory neurotransmitter in the central nervous system, and its actions are mediated by subtypes of GABA receptors named as $GABA_A$, $GABA_B,\;and\;GABA_C,\;GABA_A$, receptor consisting of ${\alpha},\;{\beta},\;{\gamma}\;and\;{\delta}$ subunits is a heterooligomeric ligand-gated chloride channel. This study was performed to investigate regulation of $GABA_A$ receptor by protein kinase C(PKC). Ion currents were recorded using gramicidine-perforated patch and whole cell patch clamp. mRNA encoding the subunits of PKC expressed in major pelvic ganglion (MPG) neurons was detected by using RT-PCR. The GABA-induced inward current was increased by PKC activators and decreased by PKC inhibitors, respectively. These effects were not associated with intracellular $Ca^{2+}$ and GAG (1-oleoyl-2-acetyl-sn-glycerol), a membrane permeable diacylglycerol (DAG) analogue. These results mean that the subfamily of PKC participating in activation of $GABA_A$ receptor would be an atypical PKC (aPKC). Among theses, ${\xi}$ isoform of aPKC was detected by RT-PCR. Taking together, we suggest that excitable $GABA_A$ receptor in sympathetic MPG neuron seemed to be regulated by aPKC, particular in ${\xi}$ isoform. The regulatory roles of PKC on excitatory $GABA_A$ receptors in sympathetic neurons of MPG may be an important factor to control the functional activity of various pelvic organs such as bowel movement, micturition and erection.

• The Journal of Korean Obstetrics and Gynecology
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• v.20 no.2
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• pp.83-96
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• 2007

Purpose: Ligustri Lucidi Fructus and Ecliptae Herba has long been used for clinical therapy associated especially with menopausal symptoms in Korea. To provide a scientific rationale for such use, we have investigated the antioxidant and vasorelaxant effects of Ligustri Lucidi Fructus, Ecliptae Herba and its mixture. Methods: The antioxidant activity of the extracts from Ligustri Lucidi Fructus, Ecliptae Herba and its mixture were evaluated and compared with that of BHA (butylated hydroxyanisole), BHT (butylated hydroxytoluene), vitamin C and vitamin E, using the ${\alpha}$,${\alpha}-diphenyl-{\beta}-picrylhydrazyl$ (DPPH) radical scavenging method. Results: Antioxidant activity of all extracts using the DPPH radical scavenging method decreased in the order vitamin C>BHA>vitamin E>Ligustri Lucidi Fructus>Ligustri Lucidi Fructus:Ecliptae Herba(1:1)>Ecliptae Herba>BHT. The vasorelaxant effects of extracts were investigated on the vasomotor tone of the rat thoracic aorta in an organ bath. All of the extracts elicited along-term relaxing response in the endothelium-intact as well as endothelium-denuded rat aorta contracted with norepinephrine. This relaxant effect was abolished by Precontraction with 72 mM KCI. Thus, it is suggested that the mechanism of vasorelaxant effect of extracts not involve voltage-operated $Ca^{2+}$ channel blocking but receptor-mediated route. Conclusion: These antioxidant and vasorelaxant effecs of the extracts may contribute to the beneficial effects in postmenopausal women.