• The Journal of Internal Korean Medicine
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• v.22 no.4
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• pp.537-546
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• 2001

목적: 고양이 대뇌피질 절편을 사용하여 저산소 발작을 유발한 뒤, 성향정기산이 세포의 이온 환경과 대사의 변화와 관련하여 어떤 영향을 미치는지 연구하였다. 방법: 고양이의 대뇌 피질 절편에 저산소 발작을 유발한 뒤 flame photometry scintillation, Spectrophotometry, method of Jorgensen and Skou, method of Fiske and Subbarow, oxygen monitor, luciferin-luciferase assay 등을 이용하여 세포내 이온함량과 세포대사를 측정하였다. 결과: 성향정기산은 저산소증으로 유발된 세포내의 $K^+$와 $Na^+$의 함량의 변화를 현저하게 지연시켰다. 성향정기산은 Na-K-ATPase의 억제제인 와바인 또는 대사억제제인 2.4-DNP로 유발된 세포내 $K^+$함량의 변화에 어떤 효과도 보이지 않았다. 또한, 정상 상태의 절편뿐만 아니라 저산소 상태의 절편에서 분리된 과립체의 분설에 있어서 Na-K-ATPase의 활동도에 영향을 미치지 않았다. 성향정기산은 저산소 발작하에서 산소 소비량과 세포의 ATP함량이 떨어지는 것을 현저하게 막았다. 또한 ATP를 생산하는 기능을 보호하는 저산소 조직의 사립체를 돕는데 효과적이었다. 결론: 성향정기산은 대뇌 조직의 저산소 발작하에서 세포의 이온 환경과 대사를 보호하는 유익한 효과가 있음을 알 수 있다.

• The Korean Journal of Physiology and Pharmacology
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• v.11 no.2
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• pp.37-43
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• 2007

Adenosine has been reported to provide cytoprotection in the central nervous systems as well as myocardium by activating cell surface adenosine receptors. However, the exact target and mechanism of its action still remain controversial. The present study was performed to examine whether adenosine has a protective effect against $A{\beta}$-induced injury in neuroglial cells. The astrocyte-derived human neuroglioma cell line, A172 cells, and $A{\beta}_{25{\sim}35}$ were employed to produce an experimental $A{\beta}$-induced glial cell injury model. Adenosine significantly prevented $A{\beta}$-induced apoptotic cell death. Studies using various nucleotide receptor agonists and antagonists suggested that the protection was mediated by $A_1$ receptors. Adenosine attenuated $A{\beta}$-induced impairment in mitochondrial functional integrity as estimated by cellular ATP level and MTT reduction ability. In addition, adenosine prevented $A{\beta}$-induced mitochondrial permeability transition, release of cytochrome c into cytosol and subsequent activation of caspase-9. The protective effect of adenosine disappeared when cells were pretreated with 5-hydroxydecanoate, a selective blocker of the mitochondrial ATP-sensitive $K^+$ channel. In conclusion, therefore we suggest that adenosine exerts protective effect against $A{\beta}$-induced cell death of A172 cells, and that the underlying mechanism of the protection may be attributed to preservation of mitochonarial functional integrity through opening of the mitochondrial ATP-sensitive $K^+$ channels.

• Proceedings of the Korean Biophysical Society Conference
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• 2003.06a
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• pp.34-34
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• 2003

Protein unfolding is a key step in several cellular processes, including protein translocation across some membranes and protein degradation by ATP-dependent proteases. C1pAP protease and the proteasome can actively unfold proteins in a process that hydrolyzes ATP, These proteases catalyze unfolding by processively unraveling their substrates from the attachment point of the degradation signal. As a consequence, the ability of a protein to be degraded depends on its structure as well as its stability. An ${\alpha}$-helix is easier to unravel than a ${\beta}$-strand. In multidomain proteins, independently stable domains are unfolded sequentially. The steric constraints imposed on substrate proteins during their degradation by the proteasome were investigated by constructing a model protein in which specific parts of the polypeptide chain were covalently connected through disulfide bridges. The cross-linked model proteins were fully degraded by the proteasome, but two or more cross-links retarded the degradation slightly. Our results suggest that the pore of the proteasome allows the concurrent passage of at least three stretches of a polypeptide chain, and also explain the limited degradation by the proteasome that occurs in the processing of the transcription factor NF-KB, and also implicate difficulty in degradation of amyloidal aggregates by the proteasome

• Microbiology and Biotechnology Letters
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• v.49 no.1
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• pp.57-64
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• 2021

Lactic acid bacteria (LAB), belonging to the Firmicutes phylum, lack heme biosynthesis and, thus, are characterized as fermentative and catalase-negative organisms. To verify the hypothesis that heme-rich-nutrients might compensate the heme-biosynthesis incapability of non-respiratory LAB in animal gut, a heme-rich-nutrient was fed to a dog and its fecal microbiome was analyzed. Firmicutes abundance in the feces from the heme-rich-nutrient-fed dog was 99%, compared to 92% in the control dog. To clarify the reason of increased Firmicutes abundance in the feces from the heme-rich-nutrient-fed dog, Lacobacillus gasseri were used as model anerobic LAB to study a purified heme (hemin). The anaerobic growth of L. gasseri in the medium with 25 µM hemin supplementation was faster than that in the medium without hemin, while the growth in the 50 µM hemin-supplemented medium did not vary. Cellular activities of the cytochrome bd complex were 1.55 ± 0.19, 2.11 ± 0.14, and 2.20 ± 0.08 U/gcell in the cells from 0, 25, and 50 µM hemin-supplemented medium, while intracellular ATP concentrations were 7.90 ± 1.12, 11.95 ± 0.68, and 12.56 ± 0.58 µmol_ATP/g_cell, respectively. The ROS-scavenging activities of the L. gasseri cytosol from 25 µM and 50 µM hemin-supplemented medium were 68% and 82% greater than those of the cytosol from no hemin supplemented-medium, respectively. These findings indicate that external hemin could compensate the heme-biosynthesis incapability of L. gasseri by increasing the cytosolic ROS-scavenging and extra ATP generation, possibly through increasing the electron transfer. Increase in the number of anaerobic bacteria in heme-rich-nutrient-fed animal gut is discussed based on the results.

• Journal of Veterinary Science
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• v.23 no.5
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• pp.76.1-76.14
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• 2022

Background: Clinical dexamethasone (DEX) treatment or stress in bovines results in extensive physiological changes with prominent hyperglycemia and neutrophils dysfunction. Objectives: To elucidate the effects of DEX treatment in vivo on cellular energy status and the underlying mechanism in circulating neutrophils. Methods: We selected eight-month-old male bovines and injected DEX for 3 consecutive days (1 time/d). The levels of glucose, total protein (TP), total cholesterol (TC), and the proinflammatory cytokines interleukin (IL)-1β, IL-6 and tumor necrosis factor (TNF)-α in blood were examined, and we then detected glycogen and adenosine triphosphate (ATP) content, phosphofructosekinase-1 (PFK1) and glucose-6-phosphate dehydrogenase (G6PDH) activity, glucose transporter (GLUT)1, GLUT4, sodium/glucose cotransporter (SGLT)1 and citrate synthase (CS) protein expression and autophagy levels in circulating neutrophils. Results: DEX injection markedly increased blood glucose, TP and TC levels, the Ca²⁺/P⁵⁺ ratio and the neutrophil/lymphocyte ratio and significantly decreased blood IL-1β, IL-6 and TNF-α levels. Particularly in neutrophils, DEX injection inhibited p65-NFκB activation and elevated glycogen and ATP contents and SGLT1, GLUT1 and GR expression while inhibiting PFK1 activity, enhancing G6PDH activity and CS expression and lowering cell autophagy levels. Conclusions: DEX induced neutrophils glucose uptake by enhancing SGLT1 and GLUT1 expression and the transformation of energy metabolism from glycolysis to pentose phosphate pathway (PPP)-tricarboxylic acid (TCA) cycle. This finding gives us a new perspective on deeper understanding of clinical anti-inflammatory effects of DEX on bovine.

• Journal of Life Science
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• v.27 no.11
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• pp.1349-1356
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• 2017

Potassium channel openers (KCOs) produce physiological and pharmacological defense mechanisms against cell injuries caused by oxidative stress of diverse origins. Openings of mitochondrial and plasmalemmal $K^+$ channels are involved in the defense mechanisms. This study tested whether NS 1619, an opener of large-conductance BK channels, has a similar beneficial influence on the pigment epithelial cells of retinas. The human retinal pigment epithelial cell line ARPE-19 was exposed to $H_2O_2$-induced oxidative stress in the absence and presence of NS 1619. The degrees of the cells' injuries were assessed by analyzing the cells' trypan-blue exclusion abilities and TUNEL staining. NS 1619 produced remarkable protections against cell injuries caused by $H_2O_2$. It prevented apoptotic and necrotic cell deaths. The protective effect of NS 1619 was significantly diminished when the cells were treated with NS 1619 in combination with the BK channel-blocker paxilline. NS 1619 significantly ameliorated cellular ATP deprivations in $H_2O_2$-treated cells. It helped mitochondria preserve their functional integrity, which was estimated by their MTT reduction abilities and mitochondrial membrane potential. In conclusion, it was suggested that NS 1619 had a beneficial effect on mitochondria in regards to preserving their functional integrity under oxidative stress, and it produces defense mechanisms against oxidant-induced cell injuries in ARPE-19 cells.

• Molecules and Cells
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• v.20 no.2
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• pp.235-240
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• 2005

Extracts of pine needles (Pinus densiflora Sieb. et Zucc.) have diverse physiological and pharmacological actions. In this study we show that pine needle extract alters pacemaker currents in interstitial cells of Cajal (ICC) by modulating ATP-sensitive $K^+$ channels and that this effect is mediated by prostaglandins. In whole cell patches at $30^{\circ}C$, ICC generated spontaneous pacemaker potentials in the current clamp mode (I = 0), and inward currents (pacemaker currents) in the voltage clamp mode at a holding potential of -70 mV. Pine needle extract hyperpolarized the membrane potential, and in voltage clamp mode decreased both the frequency and amplitude of the pacemaker currents, and increased the resting currents in the outward direction. It also inhibited the pacemaker currents in a dose-dependent manner. Because the effects of pine needle extract on pacemaker currents were the same as those of pinacidil (an ATP-sensitive $K^+$ channel opener) we tested the effect of glibenclamide (an ATP-sensitive $K^+$ channels blocker) on ICC exposed to pine needle extract. The effects of pine needle extract on pacemaker currents were blocked by glibenclamide. To see whether production of prostaglandins (PGs) is involved in the inhibitory effect of pine needle extract on pacemaker currents, we tested the effects of naproxen, a non-selective cyclooxygenase (COX-1 and COX-2) inhibitor, and AH6809, a prostaglandin EP1 and EP2 receptor antagonist. Naproxen and AH6809 blocked the inhibitory effects of pine needle extract on ICC. These results indicate that pine needle extract inhibits the pacemaker currents of ICC by activating ATP-sensitive $K^+$ channels via the production of PGs.

• Proceedings of the Ginseng society Conference
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• 1980.09a
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• pp.71-76
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• 1980

Aortic strips were prepared from rabbits, and the tensions were maintained by administration of norepinephrine into the incubation chamber. The application of diol or triol induced relaxation of the aortic strip, as indicated by the decreased aortic tension. Triol, in a concentration of $30\;mg\%\;causes\;approximately\;50\%$ of muscle relaxation, whereas a similar degree of relaxation is induced by $50\;mg\%$ of diol. This indicates that both triol and diol cause relaxation of the aorta, but that triol is about $170\%$ more potent than diol. It is well established that blood-vessel smooth-muscle tone is regulated by the available intracellular $Ca^{++}$ concentration, which in turn is profoundly influenced by interaction of the cellular membrane and sarcoplasmic reticulum in the smooth muscle. Thus, any agent which modifies the smooth-muscle tone is expected to interfere with the $Ca^{++}$ binding or uptake of sarcolemma and sarcoplasmic reticulum. In the following experiments sarcoplasmic reticulum and sarcolemma were prepared from the ventricle of rabbit heart, and the active $Ca^{++}$ uptake by these cellular components was measured employing $Ca^{45}$ in the presence of triol and diol. It was found that the active $Ca^{++}$ uptake in the presence of ATP by sarcoplasmic reticulum was inhibited by both triol and diol. Panaxatriol, in a concentration of $80\;mg\;\%,$ inhibited $Ca^{++}$ uptake by $30\%,$ whereas panaxatriol in the same concentration inhibited uptake by $20\%.$ It is clear that triol is a more potent inhibitor of active $Ca^{++}$ transport in sarcoplasmic reticulum than diol. The $Ca^{++}$ binding of the cellular membrane was also studied employing Ca45 and milipore techniques. It was found that triol in a concentration of $80\;mg\;\%,$ decreased $Ca^{++}$ binding by $29\%.$ Diol in the same concentration decreased the binding by $17\%.$ It is clear that both triol and diol inhibit $Ca^{++}$ binding to the cellular membrane, but triol is approximately $180\%$ more potent than diol.

• Molecules and Cells
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• v.43 no.1
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• pp.48-57
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• 2020

The microalga Chlamydomonas reinhardtii accumulates triacylglycerols (TAGs) in lipid droplets under stress conditions, such as nitrogen starvation. TAG biosynthesis occurs mainly at the endoplasmic reticulum (ER) and requires fatty acid (FA) substrates supplied from chloroplasts. How FAs are transferred from chloroplast to ER in microalgae was unknown. We previously reported that an Arabidopsis thaliana ATP-binding cassette (ABC) transporter, AtABCA9, facilitates FA transport at the ER during seed development. Here we identified a gene homologous to AtABCA9 in the C. reinhardtii genome, which we named CrABCA2. Under nitrogen deprivation conditions, CrABCA2 expression was upregulated, and the CrABCA2 protein level also increased. CrABCA2 knockdown lines accumulated less TAGs and CrABCA2 overexpression lines accumulated more TAGs than their untransformed parental lines. Transmission electron microscopy showed that CrABCA2 was localized in swollen ER. These results suggest that CrABCA2 transports substrates for TAG biosynthesis to the ER during nitrogen starvation. Our study provides a potential tool for increasing lipid production in microalgae.

• Journal of Ginseng Research
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• v.44 no.2
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• pp.341-349
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• 2020

Background: The replicative senescence of human dermal fibroblasts (HDFs) is accompanied by growth arrest. In our previous study, the treatment of senescent HDFs with Rg3(S) lowered the intrinsic reactive oxygen species (ROS) levels and reversed cellular senescence by inducing peroxiredoxin-3, an antioxidant enzyme. However, the signaling pathways involved in Rg3(S)-induced senescence reversal in HDFs and the relatedness of the stereoisomer Rg3(R) in corresponding signaling pathways are not known yet. Methods: We performed senescence-associated β-galactosidase and cell cycle assays in Rg3(S)-treated senescent HDFs. The levels of ROS, adenosine triphosphate (ATP), and cyclic adenosine monophosphate (cAMP) as well as the mitochondrial DNA copy number, nicotinamide adenine dinucleotide (NAD)⁺/1,4-dihydronicotinamide adenine dinucleotide (NADH) ratio, and NAD-dependent sirtuins expression were measured and compared among young, old, and Rg3(S)-pretreated old HDFs. Major signaling pathways of phosphatidylinositol 3-kinase/Akt, 5' adenosine monophosphate-activated protein kinase (AMPK), and sirtuin 1/3, including cell cycle regulatory proteins, were examined by immunoblot analysis. Results: Ginsenoside Rg3(S) reversed the replicative senescence of HDFs by restoring the ATP level and NAD⁺/NADH ratio in downregulated senescent HDFs. Rg3(S) recovered directly the cellular levels of ROS and the NAD⁺/NADH ratio in young HDFs inactivated by rotenone. Rg3(S) mainly downregulated phosphatidylinositol 3-kinase/Akt through the inhibition of mTOR by cell cycle regulators like p53/p21 in senescent HDFs, whereas Rg3(R) did not alter the corresponding signaling pathways. Rg3(S)-activated sirtuin 3/PGC1α to stimulate mitochondrial biogenesis. Conclusion: Cellular molecular analysis suggests that Rg3(S) specifically reverses the replicative senescence of HDFs by modulating Akt-mTOR-sirtuin signaling to promote the biogenesis of mitochondria.