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The Effects of Ginseng Saponin Fraction on Growth and Siderophore Formation in Eseherichia coli K-12 (인삼사포닌 분획이 Escherichia coli K-12의 성장과 Siderophore 생성에 미치는 영향)

  • 조영동;이용범
    • Journal of Ginseng Research
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    • v.7 no.2
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    • pp.102-107
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    • 1983
  • The effects of saponin, one of major components (Panax ginseng C.A. Meyer), on the growth of E. coli K-12 and the formation of siderphore was observed The following results were obtained. 1. When E. coli was grown on medium containing 1${\times}$10-5%-11${\times}$10-1% of the saponin, the rate of growth was stimulated at 10-1% of the saponin significantly compared to that of control. 2. When E. coli K-12 was grown on medium containing 1${\times}$10-1% of the saponin, the amount of siderphore was two times as much as the control. 3. The growth of E. coli was observed to be dependent on the concentration of siderophore when siderophore was added to medium. 4. The effect of saponin on the formation of siderophore in vitro was observed to reach maximum at 1${\times}$10-3% of the saponin. Such results suggest that the growth rate of E. coli K-12 could be enhanced by ginseng saponin fraction through stimulation of siderphore formation. We have described the fast growth of E. coli, K-12 and B. subtilis, rapid uptake of 14C-glucose, and high level of other metabolites such as lipids and proteins of E. coli, and B. subtilis in medium containing saponing fraction compared to that of microorganisms without saponin fraction.1∼3Such differences were claimed to be due to rapid uptake of 14C-glucose by widened periplasmic region throught unknown mechanism in the prescence of saponin fraction in medium3 and have raised a question whether there is another possible factor, siderophore4(Greek for iron bears), since microorganisms must secure a sufficient amount of iron for normal growth. These are known to be synthesized by the cells under iron-deficient condition and in most case, excreted into the medium5, where they can complex and solubilize any iron present there. It is generally believed that these complexes are then taken into the cells presumably by specific transport systems, thus providing iron for cell metabolism. Within the group of enteric bacteria, only three species (E. coli, S. typhimurium, and A. aerogense) have, so far, been studied in a ny detail. The main iron-binding compound produced by these species is enterochelin, and its role in iron transport is now well established. And biosynthesis of enterochelin from 2, 3- dihydroxybenzoate and serine in the prescence of magnesium ions and ATP was reported6. 2, 3-dihydroxybenzoate was also shown to involve isochorismate and 2, 3-dihydro-2, 3-dihydroxybenzoate as intermediate.7∼11 The present paper deals with the effect of ginseng saponin fraction on growth, the level of enterochelin formation in vivo and the conversion of 2, 3-dihydroxybenzoate and serine into entrochelin in vitro, and entrochelin obtained on the growth in relation to possible explanation of ginseng saponin fraction on the rapid growth of E. coli, K-12.

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Effect of Packaging Method on Physico-Chemical Properties of Chilled Plaice Muscle (포장 방법이 냉장 광어육의 물리$\cdot$화학적 변화에 미치는 영향)

  • Shin Wan-Chul;Song Jae-Chul;Choe Suck-Young
    • The Korean Journal of Food And Nutrition
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    • v.18 no.3
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    • pp.168-174
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    • 2005
  • Various changes of physico-chemical characteristics of chilled plaice muscle during storage at $4^{\circ}C$ in vacuum and aerobic packaging methods were examined. As a storaging period become longer, Hunter L, a, b value changes slightely. However, no differences were observed between vacuum and aerobic packaging method. The hardness of plaice muscle after death was $2,232\;dyne/cm^2$. The hardness of vacuum packaged plaice muscle storaged for 4 days was similar to that of aerobic packaged plaice muscle storaged fur 14 days. MFI(Myofibrillar Fragmentation Index) of aerobic and vacuum packaged plaice muscle showed maximum value at storage for 4days and 7 days, respectively. Mg-ATPase activities of mypofibril were increased gradually both of all during storage days. But that of MF from aerobic packaging plaice muscle was higher than that of vacuum packaging plaice muscle.

Identification of Genes Modulated by High Extracellular Calcium in Coculture of Mouse Osteoblasts and Bone Marrow Cells by Oligo Chip Assay

  • Kim, Hyung-Keun;Song, Mi-Na;Jun, Ji-Hae;Woo, Kyung-Mi;Kim, Gwan-Shik;Baek, Jeong-Hwa
    • International Journal of Oral Biology
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    • v.31 no.2
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    • pp.53-65
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    • 2006
  • Calcium concentration in the bone resorption lacunae is high and is in the mM concentration range. Both osteoblast and osteoclast have calcium sensing receptor in the cell surface, suggesting the regulatory role of high extracellular calcium in bone metabolism. In vitro, high extracellular calcium stimulated osteoclastogenesis in coculture of mouse osteoblasts and bone marrow cells. Therefore we examined the genes that were commonly regulated by both high extracellular calcium and $1,25(OH)_2vitaminD_3(VD3)$ by using mouse oligo 11 K gene chip. In the presence of 10 mM $[Ca^{2+}]e$ or 10 nM VD3, mouse calvarial osteoblasts and bone marrow cells were co-cultured for 4 days when tartrate resistant acid phosphatase-positive multinucleated cells start to appear. Of 11,000 genes examined, the genes commonly regulated both by high extracellular calcium and by VD3 were as follows; 1) the expression of genes which were osteoclast differentiation markers or were associated with osteoclastogenesis were up-regulated both by high extracellular calcium and by VD3; trap, mmp9, car2, ctsk, ckb, atp6b2, tm7sf4, rab7, 2) several chemokine and chemokine receptor genes such as sdf1, scya2, scyb5, scya6, scya8, scya9, and ccr1 were up-regulated both by high extracellular calcium and by VD3, 3) the genes such as mmp1b, mmp3 and c3 which possibly stimulate bone resorption by osteoclast, were commonly up-regulated, 4) the gene such as c1q and msr2 which were related with macrophage function, were commonly down-regulated, 5) the genes which possibly stimulate osteoblast differentiation and/or mineralization of extracellular matrix, were commonly down-regulated; slc8a1, admr, plod2, lox, fosb, 6) the genes which possibly suppress osteoblast differentiation and/or mineralization of extracellular matrix, were commonly up-regulated; s100a4, npr3, mme, 7) the genes such as calponin 1 and tgfbi which possibly suppress osteoblast differentiation and/or mineralization of extracellular matrix, were up-regulated by high extracellular calcium but were down-regulated by VD3. These results suggest that in coculture condition, both high extracellular calcium and VD3 commonly induce osteoclastogenesis but suppress osteoblast differentiation/mineralization by regulating the expression of related genes.

Hepatic microRNAome reveals potential microRNA-mRNA pairs association with lipid metabolism in pigs

  • Liu, Jingge;Ning, Caibo;Li, Bojiang;Li, Rongyang;Wu, Wangjun;Liu, Honglin
    • Asian-Australasian Journal of Animal Sciences
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    • v.32 no.9
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    • pp.1458-1468
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    • 2019
  • Objective: As one of the most important metabolic organs, the liver plays vital roles in modulating the lipid metabolism. This study was to compare miRNA expression profiles of the Large White liver between two different developmental periods and to identify candidate miRNAs for lipid metabolism. Methods: Eight liver samples were collected from White Large of 70-day fetus (P70) and of 70-day piglets (D70) (with 4 biological repeats at each development period) to construct sRNA libraries. Then the eight prepared sRNA libraries were sequenced using Illumina next-generation sequencing technology on HiSeq 2500 platform. Results: As a result, we obtained 346 known and 187 novel miRNAs. Compared with the D70, 55 down- and 61 up-regulated miRNAs were shown to be significantly differentially expressed (DE). Gene ontology and Kyoto encyclopedia of genes and genomes enrichment analysis indicated that these DE miRNAs were mainly involved in growth, development and diverse metabolic processes. They were predicted to regulate lipid metabolism through adipocytokine signaling pathway, mitogen-activated protein kinase, AMP-activated protein kinase, cyclic adenosine monophosphate, phosphatidylinositol 3 kinase/protein kinase B, and Notch signaling pathway. The four most abundantly expressed miRNAs were miR-122, miR-26a and miR-30a-5p (miR-122 only in P70), which play important roles in lipid metabolism. Integration analysis (details of mRNAs sequencing data were shown in another unpublished paper) revealed that many target genes of the DE miRNAs (miR-181b, miR-145-5p, miR-199a-5p, and miR-98) might be critical regulators in lipid metabolic process, including acyl-CoA synthetase long chain family member 4, ATP-binding casette A4, and stearyl-CoA desaturase. Thus, these miRNAs were the promising candidates for lipid metabolism. Conclusion: Our study provides the main differences in the Large White at miRNA level between two different developmental stages. It supplies a valuable database for the further function and mechanism elucidation of miRNAs in porcine liver development and lipid metabolism.

NLRP3 Inflammasome in Neuroinflammatory Disorders (NLRP3 인플라마좀 작용 기전 및 신경 질환에서의 역할)

  • Kim, Ji-Hee;Kim, YoungHee
    • Journal of Life Science
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    • v.31 no.2
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    • pp.237-247
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    • 2021
  • Immune responses in the central nervous system (CNS) function as the host's defense system against pathogens and usually help with repair and regeneration. However, chronic and exaggerated neuroinflammation is detrimental and may create neuronal damage in many cases. The NOD-, LRR-, and pyrin domain―containing 3 (NLRP3) inflammasome, a kind of NOD-like receptor, is a cytosolic multiprotein complex that consists of sensors (NLRP3), adaptors (apoptosis-associated speck like protein containing a caspase recruitment domain, ASC) and effectors (caspase 1). It can detect a broad range of microbial pathogens along with foreign and host-derived danger signals, resulting in the assembly and activation of the NLRP3 inflammasome. Upon activation, NLRP3 inflammasome leads to caspase 1-dependent secretion of the pro-inflammatory cytokines IL-1β and IL-18, as well as to gasdermin D-mediated pyroptotic cell death. NLRP3 inflammasome is highly expressed in CNS-resident cell types, including microglia and astrocytes, and growing evidence suggests that NLRP3 inflammasome is a crucial player in the pathophysiology of several neuroinflammatory and psychiatric diseases, such as Alzheimer's disease, Parkinson's disease, multiple sclerosis, stroke, traumatic brain injury, amyotrophic lateral sclerosis, and major depressive disorder. Thus, this review describes the molecular mechanisms of NLRP3 inflammasome activation and its crucial roles in the pathogenesis of neurological disorders.

Effect of Cadmium on the Expression of ABC Transporters and Glutathione S-transferase in the Marine Ciliate Euplotes crassus (카드뮴이 해양 섬모충(Euplotes crassus)의 ABC Transporters와 GST 유전자 발현에 미치는 영향에 관한 연구)

  • Kim, Hokyun;Kim, Se-Hun;Kim, Ji-Soo;Lee, Young-Mi
    • Journal of Marine Life Science
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    • v.1 no.2
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    • pp.79-87
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    • 2016
  • Heavy metals such as cadmium (Cd) are highly toxic to aquatic organisms and human, even at trace concentration. Herein we investigated the effect of Cd on the gene expression of ATP-binding cassette (ABC) transporters and glutathione S-transferase (GST) in marine ciliate Euplotes crassus. Seven ABC transporters and one GST genes were partially cloned and sequences, and thereafter, transcriptional modulation of these genes after exposure to Cd for 8 h was investigated using quantitative real time RT- PCR (qRT-PCR). As results, sequence analysis and phylogenetic study revealed that E. crassus ABCs are likely typical ABC transports, in particular, B/C family, and GST gene may be similar to GST theta isoform. A significant increase in the expression of ABCs, except for ABCB21 was observed in a concentration dependent manner after exposure to Cd (0.1 and 0.5 mg/l) for 8 h. The GST mRNA level was the highest at 0.5 mg/l Cd and then reduced until control level. These findings suggest that ABCs and GST may be involved in a protective mechanism against Cd-mediated toxicity in E. crassus.

Mitigation Effects of Foliar-Applied Hydrogen Peroxide on Drought Stress in Sorghum bicolor (과산화수소 엽면 처리에 의한 수수에서 한발 스트레스 완화 효과)

  • Shim, Doo-Do;Lee, Seung-Ha;Chung, Jong-Il;Kim, Min Chul;Chung, Jung-Sung;Lee, Yeong-Hun;Jeon, Seung-Ho;Song, Gi-Eun;Shim, Sang-In
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.65 no.2
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    • pp.113-123
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    • 2020
  • Global climatic change and increasing climatic instability threaten crop productivity. Due to climatic change, drought stress is occurring more frequently in crop fields. In this study, we investigated the effect of treatment with hydrogen peroxide (H2O2) before leaf development on the growth and yield of sorghum for minimizing the damage of crops to drought. To assess the effect of H2O2 on the growth of sorghum plant, 10 mM H2O2 was used to treat sorghum leaves at the 3-leaf stage during growth in field conditions. Plant height, stem diameter, leaf length, and leaf width were increased by 7.6%, 9.6%, 8.3% and 11.5%, respectively. SPAD value, chlorophyll fluorescence (Fv/Fm), photosynthetic rate, stomatal conductance, and transpiration rate were increased by 3.0%, 4.9%, 26.0%, 23.4% and 12.7%, respectively. The amount of H2O2 in the leaf tissue of sorghum plant treated with 10 mM H2O2 was 0.7% of the applied amount after 1 hour. The level increased to approximately 1.0% after 6 hours. The highest antioxidant activity measured by the Oxygen Radical Absorbance Capacity assay was 847.3 µmol·g-1 at 6 hour after treatment. However, in the well-watered condition, the concentration of H2O2 in the plant treated by the foliar application of H2O2 was 227.8 µmol·g-1 higher than that of the untreated control. H2O2 treatment improved all the yield components and yield-related factors. Panicle length, plant dry weight, panicle weight, seed weight per plant, seed weight per unit area, and thousand seed weight were increased by 8.8%, 18.0%, 24.4%, 24.7%, 29.9% and 7.1%, respectively. Proteomic analysis showed that H2O2 treatment in sorghum increased the tolerance to drought stress and maintained growth and yield by ameliorating oxidative stress.

Study of Hedyotis Diffusa Methanol Extract on Anti-tumoral Effect and Mechanism (백화사설초(白花蛇舌草) 메탄올 추출물(抽出物)의 항종양(抗腫瘍) 효과(效果) 및 항암(抗癌) 기전(機轉)에 관(關)한 연구(硏究))

  • No, Hoon-Jeong;Moon, Gu;Moon, Seok-Jae;Won, Jin-Hee;Moon, Young-Ho;Park, Rae-Gil
    • THE JOURNAL OF KOREAN ORIENTAL ONCOLOGY
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    • v.6 no.1
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    • pp.81-97
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    • 2000
  • Objectives: This experimental study was carried out to evaluate the effects of aqueous and methanol extracts of Hedyotis diffusa which has long been used for cancer treatment in oriental medicines on the induction of apoptotic cell death in human lymphoid leukemia cell line, HL-60. Methods: Cells were treated with various concentrations (200 to $0.4{\mu}g$) and periods (6 to 30 hr) of $H_2O$ and methanol extracts of Hedyotis diffusa. Then, cells were tested for viability by MTT assay. Cells wrere treated with $200{\mu}g/ml$ of methanol extract fork various periods. Genomic DNA was isolated, separated, on 1.5% agarose gels, stained with ethidium bromide and visualized under UV light. Cells were treated with $200{\mu}g/ml$ of each extract for 16 hr. Then, cells were treated with Hoechst dye 33342 and observed by fluorescence microscopy. Cells were treated with various doses of each for 12 hr and $100{\mu}g/ml$ of methanol extract for various periods. Lysate from the cells used to measure the activity of Caspase-1 and-3 proteases by using fluorogenic peptide substrates including acetyl-YVAD-AMC and acetyl-DEVD-AMC, respectively. Cells were treated with $200{\mu}g/ml$ of each extract for various periods. Cell lysates were immunoprecipated with anti-JNKl antibodies. The immune complex was reacted with $32^p-ATP$ and c-Jun as a substrate. The phosphotransferase activity of JNKI was measured by using PhosphoImage analyzer (Fuji Co., Japan). Nuclear extracts were isolated and incubated with oligonucleotide probe of $NF-{\kappa}B$. Transcriptional activation of ${\kappa}B$ was measured by using EMSA and visualized by PhosphoImage analyzer (Fuji Co, Japan). Cell lysates were prepared and analyzed by Western blotting with anti-Bc12 antibodies and anti-Bax antibodies. Cells were pretreated with various doses of methanol extract for 2 hr. Then, the extract was removed by centrifugation. Cells were resuspended with RPMI-1640 media containing 0.3% agarose, 10% FBS, overlayred onto bottom layer agarose and incubated at $CO_2$ incubator for 6 days. The number of colony was counted under light microscopy ($\time100$). Results: The death of HL-60 cells was markedly induced by the addition of methanol extract of Hedyotis diffusa in a dose and time-dependent manners. The apoptotic characteristic ladder pattern of DNA strand break was observed in death of HL-60 cells. In addition, it was shown nucleus chromatin condensation and fragmentation under Hoechst staining. Therefore, Hedyotis diffusa extract-induced death of HL-60 cells is mediated by apoptotic signaling processes. The activity of Caspase 3-like proteases remained in a basal level in HL-60 cells treated with aqueous extract of Hedyotis diffusa. However, it was markedly increased in HL-60 cells treated with methanol extract of Hedyotis diffusa. In addition, the phosphotransferase activity of JNKl was increased in HL-60 cells treated with methanol extract of Hedyotis diffusa. Furthermore, the activation of transcriptional activator, $NF-{\kappa}B$ was markedly induced by methanol extract of Hedyotis diffusa. Anti-apoptotic Bc12 was cleaved into 23Kda fragment by treatment of methanol extract of Hedyotis diffusa. However, expression of proapoptotic Bax protein was increased by treatment of methanol extract of Hedyotis diffusa in a time-dependent manner. Furthermore, methanol extract markedly inhibited the colony forming efficiency of HL-60 cells in semisolid agar culture. Conclusions: Above results suggest that methanol extract of Hedyotis diffusa induces the apoptotic death of human leukemic HL-60 cells via activations of Caspase-3 proteases, JNKI, transcriptional activator $NF-{\kappa}B$, In addition, our results also suggest that methanol extract of Hedyotis diffusa reduces the malignant potential of HL-60 cells via down regulation of colony forming effciency through cleavage of Bc12 as well as induction of Bax.

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Adenyl Cyclase Activity in Cold-acclimatized Animals (한냉적응이 Adenyl Cyclase Activity에 미치는 영향)

  • Kang, Bok-Soon;Lee, Sang-Ho;Kang, Doo-Hee
    • The Korean Journal of Physiology
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    • v.8 no.2
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    • pp.67-74
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    • 1974
  • The object of this research is aimed to determine the activity of adenyl cyclase in both skeletal muscle sarcolemma and fat cell ghost of epididymal adipose tissue isolated from rats exposed to cold for various length of time in an attempt to evaluate whether the tissue sensitivity to catecholamine is increased when rats are exposed to cold for long periods of time Methods: a)Animals: Albino rats ranging in weight from 150 to 200 gm were used throughout this study. For experimental purposes, the rats are divided into two groups: experimental animals were place4 in a cold room at $4^{\circ}C$, controls being kept at $25^{\circ}C$. At the end of 2, 4, 6, 12, and 16 weeks. exposure to cold the rats were used to measure the adenyl cyclase activity. b) Isolation of plasma membrane from skeletal muscle and adipose tissue: The Plasma membrane of skeletal muscle from hind limbs of rats are prepared by the method employed by Rosenthal et at. and fat cell ghost of epididymal adipose tissue of rats by the method employed by Rodbell. c) Adenyl cyclase assay: Adenyl cyclase activity were measured by the method employed by Marinetti et al. Briefly, plasma membrane was incubated with $3^H-ATP$, various amount of noradrenaline and other incubation mixture at $37^{\circ}C$ for 20 minutes. After stopping the enzyme reaction by immersion in boiling water, carrier 3',5'-AMP was added to the system as a marker and $100\;{\mu}1$ aliquots of incubation mixture were pipetted on $20{\time}20$ Whatman No. 3 MM filter paper for one dimensional chromatography. The cyclic AMP spots were cut off and placed in counting vials containing 10ml of Bray's scintillation cocktail. Radioactivity was determined with a Packard Tri-Carb liquid scintillation counter. The enzyme activity is expressed as nanomoles of cyclic AMP produced per mg of membrane per hour. Result: 1. Average adenyl cyclase activity in the plasma membrane of skeletal muscle before and after noradrenaline administration was significantly higher in the cold-exposed rats as compared to the control. Continuous exposure to cold Produced an increased adenyl cyclase activity before and after noradrenaline administration. Adenyl cyclase activity reached peak levels at the 6 weeks exposure to told and level of adenyl cyclase activity remained high. Noradrenaline administration to the incubation medium induced a significant increase in adenyl cyclase activity and the degree of stimulation were proportional to the hormonal concentration But the rate of inclement in adenyl cyclase activity by noradreasline was the same in both groups. 2. Adenyl cyclase activity in fat cell ghost between cold exposed and control rats showed no significant differences before and after noradreualine administration. In summary, it can be concluded that cold adaptation give rise an increased activity of adenyl cyclase in plasma membrane of skeletal muscle in rats.

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Inhibitory Effects of Potassium Channel Openers on the Oxytocin-induced Contraction of the Rat Uterus in vitro (쥐자궁근의 운동성에 대한 $K^+$채널 개방제의 이완 작용)

  • Kim, Hee-Jeong;Lee, Mun-Han;Ryu, Pan-Dong
    • The Korean Journal of Pharmacology
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    • v.30 no.2
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    • pp.191-203
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    • 1994
  • $K^+$ channel openers (KCOs) are known to have a wide range of effects by opening the $K^+$ channel in plasma membranes of various smooth muscles, cardiac muscle and pancreatic ${\beta}-cell$. In the present study, we investigated the effects of 5 types of KCOs, cromakalim, RP49356, pinacidil, nicorandil and diazoxide on the contractility of isolated rat uterus. All KCOs tested inhibited the uterine contraction induced by 0.2 nM oxytocin in a dose-dependent manner. Individual KCO and its $pD_2$ values were cromakalim 6.5, RP49356 6.3, pinacidil 5.92, nicorandil 4.43 and diazoxide 4.18. The relaxant effects of KCO were inhibited by glibenclamide (0.3, 1 and $10\;{\mu}M$) with $pA_2$ values of cromakalim 6.91, RP49356 6.59, pinacidil 6.55, nicorandil 5.97 and diazoxide 6.37. In addition, the relaxant effect of cromakalim or pinacidil was antagonised by TEA, a non-selective $K^+$ channel blocker, but not by apamin. Contractions induced by low concentration of KCI (< 40 mM) were inhibited by cromakalim $(100{\mu}M)$ and nicorandil $(300{\mu}M)$, but those evoked by higher concentration (> 40 mM) of KCI were little affected. In ovariectomized rat uterus, cromakalim dose-dependently inhibited oxytocin-induced contraction and glibenclamide $(10{\mu}M)$ inhibited the relaxant effect of cromakalim with $pD_2$ and $K_B$ values of 7.48 and $1.26{\times}10^{-7}M$, respectively. In estrogen-primed rat uterus, these values were 6.51 and $1.57{\times}10^{-7}M$, respectively, indicating that the cromakalim is less effective on the estrogen-treated uterine smooth muscle. Our results suggest that the KCO-sensitive $K^+$ channels participate in the motility of uterine smooth muscle and such channels are, at least in part, under the control of estrogen. In addition, our data Indicate that the type of $K^+$ channels activated by KCO is ATP-sensitive $K^+$ channels which is blocked by glibenclamide.

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