• Title/Summary/Keyword: ATP

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The Taste Compounds in Fermented Entrails of Clupanodon Osdeckii (전어 내장(內臟)젓 의 맛성분(成分))

  • Chung, Seung-Yong;Kim, Hee-Suk
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.9 no.1
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    • pp.23-32
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    • 1980
  • This study was attempted to establish the basic data for evaluating taste compounds in fermented entrails of Clupanodon Osdeckii. The changes of such compounds as amino acids, nucleotides and their related compounds, betaine, TMAO and TMA during fermentation were analyzed. IMP, AMP, ADP and ATP were decreased, while hypoxanthine was increased during the fermentation. The content of hypoxanthine in fermented entrails of Clupanodon Osdeckii after 50 days was increased to about 2 times of that in raw entrails. In the free amino acid composition of raw entrails, abundant amino acids were lysine, glutamic acid, valine, alanine, threonine, serine, leucine and glycine in order. Such amino acids as arginine, tyrosine and phenylalanine were lower than 2.0% of total free amino acid, and proline and cysteine were detected in trace amount. The changes in free amino acid composition of the extract in entrails of Clupanodon Osdeckii during fermentation were not observed. Such amino acids as lysine, glutamic acid, valine, serine and leucine were especially abundant in both raw and fermented products. The content of total free amino acids in fermented entrails of Clupanodon Osdeckii after 50 days were increased to about 12 times of that in raw. The content of betaine nitrogen were about 14.5 (moisture and salt free base) after 50 days of fermentation. TMAO nitrogen was decreased during the fermentation. It is believed that lysine, glutamic acid, valine, serine, leucine and hypoxanthine play an important role as taste compounds in fermented entrails of Clupanodon Oseckii.

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Changes in Physicochemical Characteristics of Squid upon Acid and Heat Treatment (산 및 열처리에 따른 오징어의 이화학적 특성변화)

  • Lee, Hye-Young;Kim, Seong-Ho;Kim, Duk-Jin
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.41 no.4
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    • pp.539-549
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    • 2012
  • In order to acquire basic data on the development of squid processing food, we investigated changes in the composition of boiled squid upon heat treatment ($100^{\circ}C$), acid treatment (acetic acid, 0~5%), and pre-boiling ($55^{\circ}C$, $80^{\circ}C$). The proximate composition of squid was 73~78% moisture and 19~24% crude protein, treatment with acid solution had a significant effect on the proximate composition of boiled squid (p<0.05). The major free sugars were ribose and glucose in all treatment samples. The $55^{\circ}C$ pre-boiled sample had lower levels of glucose than the other samples. The total free sugar content of the non-peeled sample was the highest, followed by the $80^{\circ}C$ pre-boiled sample, whereas the sugar content in the $55^{\circ}C$ pre-boiled sample was very low. With regards to amino acid content, proline was the highest in all samples, followed by taurine and histidine. Treatment with acid solution had a significant effect on the total free amino content of boiled squid (p<0.05). The total free amino acid content of the $55^{\circ}C$ pre-boiled sample was the highest, followed by the $80^{\circ}C$ pre-boiled sample and non-peeled sample. Inosine and related compounds were not detected in any of the samples, and the adenosine triphosphate (ATP) content was low. The hypoxanthine contents of the $55^{\circ}C$ and $80^{\circ}C$ pre-boiled samples were the highest, the adenosine monophosphate (AMP) and inosine monophosphate (IMP) contents were similar, and the IMP content of the non-peeled sample was higher than those of the peeled samples. The palmitic acid content was very high and constituted 40% of total saturated fatty acids. eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) contents were also high and constituted 60% of total unsaturated fatty acids. Of these, DHA content was the highest, followed by palmitic acid and EPA, which accounted for about 85% of total fatty acids. No difference in fatty acid content was observed between acid treatment and pre-boiling. The mineral P content was the highest on average in all boiled squid samples, followed by K, Na, Mg, and Ca contents. In addition, the pre-boiling temperature and acid solution concentration had significant effects on the mineral content. Further, heavy metal, Cd, Pb, and As contents were detected only at trace amounts, and their levels were lower than standard and permissible amounts for food.

Changes in Physicochemical Composition of Sea Urchin Roe by Steaming Treatment (열처리 조건에 따른 성게 알의 이화학적 성분 변화)

  • Lee, Sung-Uk;Lee, Hye-Young;Kim, Seong-Ho;Kim, Duk-Jin
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.41 no.4
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    • pp.550-560
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    • 2012
  • This study was performed to investigate the physicochemical properties of sea urchin ($Anthocidaris$ $crassispina$, $Pseudocentrotus$ $depressus$, $Hernicentrotus$ $pulcherrimus$) roe as a processed or canned food by steaming treatment. Proximate compositions of $A.$ $crassispina$ roe and $P.$ $depressus$ roe were similar, but water, crude ash, and carbohydrate contents of $H.$ $pulcherrimus$ roe showed little differences. Proximate compositions of sea urchin roe showed slight differences with steaming time, raw samples showed no differences. Glycine content of the three raw sea urchin roe samples showed the highest concentration among free amino acids, followed by arginine, alanine, and lysine, in order. Total free amino acid contents of raw sea urchin roe were 754.70 mg% ($A.$ $crassispina$), 567.75 mg% ($P.$ $depressus$), and 449.44 mg% ($H.$ $pulcherrimus$). Total free amino acid content of 5 min steaming sample was highest among steaming and canning conditions. ATP, ADP, and AMP contents of raw $P.$ $depressus$ roe sample was higher than those of $A.$ $crassispina$and $H.$ $pulcherrimus$ roe. Major fatty acids of the three raw sea urchin roe samples were myristic acid, palmitic acid, and EPA. S.F.A. content of raw samples of $A.$ $crassispina$and $H.$ $pulcherrimus$ roe was higher than U.F.A content, whereas U.F.A. content of $P.$ $depressus$ roe was highest among the three raw samples. For minerals K, P, Fe, and Zn contents were highest in $A.$ $crassispina$roe while Ca, Mg, Na, and Cu contents were highest in $H.$ $pulcherrimus$ roe. For heavy metals, Cd, Pb and As were detected in all samples in trace amounts under the criteria of the Korea food codex.

Effects of Ginsenosides on Glucose Uptake and Insulin Secretion

  • Park, Min-Woo;Shin, Eun-Jung;Ko, Sung-Kwon;Chung, Sung-Hyun
    • Proceedings of the Ginseng society Conference
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    • 2007.05a
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    • pp.23-24
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    • 2007
  • Purpose: 인삼이 항당뇨 활성을 가진다는 연구가 많은 연구자들에 의해 진행되었고, 이는 인삼의 구성 성분 중 ginsenoside에 기인한다는 보고가 있다. 본 연구는 ginsenoside의 항당뇨 작용기전을 in vitro에서 알아보고자 3T3-L1 지방세포에서 glucose uptake와 췌장 베타세포인 HIT-T15 세포에서 insulin 분비 효과를 확인하였다. 이를 위하여 인삼을 식초로 처리한 긴삼의 70% MeOH 분획으로부터 protopanaxadiol 계인 ginsenoside $Rb_2$, $Rg_3$ 그리고 protopanaxtriol 계인 $Rg_2$를 분리하여 본 실험에 사용하였다. Method: Ginsenoside $Rb_2$, $Rg_2$, $Rg_3$가 지방 세포에서 glucose uptake에 미치는 효과를 확인하기 위하여 3T3-L1 세포를 DMEM (Dulbecco's Modified Eagle's Medium) 배지에서 분화 유도시켰으며 3T3-L1 preadipocyte가 80% 정도 자라면 분화 유도 배지 (5% fetal bovine serum (FBS), 0.5 mM isobutylmethylxanthine (IBMX), 1 mM dexamethasone 그리고 $10{\mu}g/ml$ insulin가 포함된 DMEM)로 4일, $10{\mu}g/ml$ insulin가 포함된 DMEM으로 2일, FBS만 포함된 DMEM으로 2일 배양하여 총 8일 동안 분화를 유도하였다. 분화 유도된 3T3-L1 adipocytes 에 각각 $Rb_2$, $Rg_2$, $Rg_3$$20{\mu}M$로 처리하여 16시간 배양하여 low glucose DMEM에서 3시간 배양한 후에 $37^{\circ}C$에서 insulin 10 ng/ml 과 각각 $Rb_2$, $Rg_2$, $Rg_3$가 포함된 Krebs Ringer Hepes buffer(KRP buffer)에서 20분간 배양하였다. 2-deoxy-D-[$^3H$]-glucose를 넣고 10분 후에 차가운 PBS로 반응을 종결시켜 lysis buffer로 cell을 모은 후 scintillation counter를 이용하여 glucose를 측정하였다. Insulin 분비 효과는 HIT-T15 세포와 일차 배양한 흰쥐 소도세포(islets)를 사용하여 확인하였다. HIT-T15 세포는 24 well plate에 well 당 $2{\times}10^5$ 개씩 분주하여 24시간 동안 배양한 후 시료를 처리하였으며 소도 세포는 Sprague-Dawley rat의 췌장에 collagenase가 포함된 Hanks' Balanced Salt Solution(HBSS)을 주입하여 분리하고 islets을 얻었다. 분리한 소도세포를 $1{\sim}2$일 동안 배양하여 $Rb_2$, $Rg_2$, $Rg_3$가 각각 $20{\mu}M$의 농도로 첨가된 insulin 측정용 buffer인 Krebs-Ringer buffer (KRB+0.3% BSA, KRBB)에 $37^{\circ}C$에서 1시간 incubation 시킨 후 배양액으로 분비된 인슐린의 양을 측정하였다. 한편 ginsenoside의 인슐린 분비 촉진 기전을 알아보기 위한 실험에서는 ATP-sensitive $K^+$ channel opener인 diazoxide (0.5 mM)가 ginsenoside에 의해 촉진된 인슐린 분비를 억제하는지 살펴보았다. Result: glucose uptake assay 에서는 $Rg_2$가 가장 크게 glucose uptake를 증가시켰고 $Rb_2$, $Rg_3$는 그 활성이 크지 않았다. 한편 Insulin 분비 효과는 diol계인 $Rg_3$에서 용량 의존적으로 인슐린의 분비를 촉진시켰으며 $20{\mu}M$ 농도에서 대조군과 비교해 1.5배 이상의 분비 촉진 효과를 보였고 triol계인 $Rg_2$ 에서는 이러한 효과가 나타나지 않았다. $Rg_3$의 인슐린 분비 촉진 기전을 0.5 mM 의 diazoxide를 이용하여 확인한 결과 $Rg_3$에 의해 촉진된 인슐린 분비를 감소시켰다. 이로 미루어보아 $Rg_3$의 인슐린 분비 촉진 기전은 ATP-sensitive $K^+$ 채널의 봉쇄에 의한 것임을 확인할 수 있었다.

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Flavor Constituents in Enzyme Hydrolysates from Shore Swimming Crab and Spotted Shrimp (민꽃게 및 꽃새우 효소가수분해물의 풍미발현성분)

  • ;;Chi-Tang Ho
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.30 no.5
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    • pp.787-795
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    • 2001
  • For the developing natural fisheries flavoring substances using crustacea, the flavor constituents of enzyme hydrolysates from shore swimming crab (crab) and spotted shrimp (shrimp) were investigated. In taste-active compounds of both enzyme hydrolysates, total free amino acid contents of crab and shrimp enzyme hydrolysates were 5,226.7 mg% and 8,757.3 mg%, respectively. The major amino acids were taurine, glutamic acid, proline, asparagine, glycine, alanine, valine, leucine, lysine anserine and arginine. As for ATP related compounds, AMP was the principal component and small amounts of IMP was detected in both enzyme hydrolysates. In the quarternary ammonium bases, betaine was the principal component (593.8mg%), and contents of TMAO and betaine in both samples were 60.7 mg% and 850.0 mg%, 124.1 mg% and 755.9 mg%, respectively. The major components were Na, K, P and Cl in inorganic ions. The major fatty acids of both sample were 14 : 0, 16 : 0, 16 : 1n7, 18 : 1n9, 20 : 5n3 and 22 : 6n3, and composition ration of n3 polyunsaturated fatty acids of were 27.8% and 28.5%, respectively. Total 99~109 volatile compounds were detected as a cooked odor of crab and shrimp enzyme hydrolysates by SDE apparatus/gas chromatography/mass spectrometry. The volatile flavor compounds identified from cooked crab enzyme hydrolysate were composed of 6 acids, 10 alcohols, 7 aldehydes, 11 ketones, 1 ester, 5 phenols, 4 benzenes, 22 hydrocarbons, 1 furan, 21 nitrogen containing compounds and 11 micellaneous compounds. And the volatile flavor compounds indentified from cooked shrimp enzyme hydrolysate were composed of 13 acids, 10 alcohols, 6 aldehydes, 10 ketones, 3 esters, 2 phenols, 5 benzenes, 36 hydrocarbons, 1 furan, 14 nitrogen containing compounds and 8 micellaneous compounds.

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Effect of Halophilic Bacterium, Haloarcula vallismortis, Extract on UV-induced Skin Change (호염 미생물(Haloarcula vallismortis) 용해물의 자외선유발 피부변화에 대한 효과)

  • Kim, Ji Hyung;Shin, Jae Young;Hwang, Seung Jin;Kim, Yun Sun;Kim, Yoo Mi;Gil, So Yeon;Jin, Mu Hyun;Lee, Sang Hwa
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.41 no.4
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    • pp.341-350
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    • 2015
  • Skin carrys out protective role against harmful outer environment assaults including ultraviolet radiation, heavy metals and oxides. Especially, ultraviolet-B (UVB) light causes inflammatory reactions in skin such as sun burn and erythma and stimulates melanin pigmentation. Furthermore, the influx of UVB into skin cells causes DNA damage in keratinocytes and dermal fibroblasts, inhibition of extracellular matrix (ECM) synthesis which leads to a decrease in elasticity of skin and wrinkle formation. It also damages dermal connective tissue and disrupts the skin barrier function. Prolonged exposure of human skin to UVB light is well known to trigger severe skin lesions such as cell death and carcinogenesis. Haloarcula vallismortis is a halophilic microorganism isolated from the Dead Sea, Its growth characteristics have not been studied in detail yet. It generally grows at salinity more than 10%, but the actual growth salinity usually ranges between 20 to 25%. Because H. vallismortis is found mainly in saltern or salt lakes, there could exist defense mechanisms against strong sunlight. One of them is generation of additional ATP using halorhodopsin which absorbs photons and produces energy by potential difference formed by opening the chloride ion channel. It often shows a color of pink or red because of their high content of carotenoid pigments and it is considered to act as a defense mechanism against intense UV irradiation. In this study, the anti-inflammatory effect of the halophilic microorganism, H. vallismortis, extract was investigated. It was found that H. vallismortis extract had protective effect on DNA damage induced by UV irradiation. These results suggest that the extract of halophilic bacterium, H. vallismortis could be used as a bio-sunscreen or natural sunscreen which ameliorate the harmful effects of UV light with its anti-inflammatory and DNA protective properties.

Protective effect of Gabjubaekmok (Diospyros kaki) extract against amyloid beta (Aβ)-induced cognitive impairment in a mouse model (아밀로이드 베타(amyloid beta)로 유도된 인지장애 마우스 모델에서 갑주백목(Diospyros kaki) 추출물의 인지기능 및 뇌 신경세포 보호 효과)

  • Yoo, Seul Ki;Kim, Jong Min;Park, Seon Kyeong;Kang, Jin Yong;Han, Hye Ju;Park, Hyo Won;Kim, Chul-Woo;Lee, Uk;Heo, Ho Jin
    • Korean Journal of Food Science and Technology
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    • v.51 no.4
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    • pp.379-392
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    • 2019
  • The current study investigated the effect of Gabjubaekmok (Diospyros kaki) ethanolic extract (GEE) on $H_2O_2$-induced human neuroblastoma MC-IXC cells and amyloid beta $(A{\beta})_{1-42}$-induced ICR (Institute of Cancer Research) mice. GEE showed significant antioxidant activity that was evaluated based on ABTS, DPPH scavenging activity, and inhibition of malondialdehyde (MDA) and acetylcholinesterase activity. Further, GEE inhibited ROS production and increased cell viability in $H_2O_2$-induced MC-IXC cells. Administration of GEE ameliorated the cognitive dysfunction on $A{\beta}$-induced ICR mice as evaluated using Y-maze, passive avoidance, and Morris water maze tests. Results of ex vivo test using brain tissues showed that, GEE protected the cholinergic system and mitochondrial functions by increasing the levels of antioxidants such as ROS, mitochondrial membrane potential (MMP), and adenosine triphosphate (ATP) against $A{\beta}$-induced cognitive dysfunction. Moreover, GEE decreasd the expression levels of apoptosis-related proteins such as $TNF-{\alpha}$, p-JNK, p-tau, BAX and caspase 3. While, expression levels of p-Akt and $p-GSK3{\beta}$ increased than $A{\beta}$ group. Finally, gallic acid was identified as the main compound of GEE using high performance liquid chromatography.

Effect of methyl jasmonate on the glucosinolate contents and whole genome expression in Brassica oleracea (유묘기 양배추류에서 메틸자스모네이트에 의한 글루코시놀레이트 함량 변화 및 전사체 발현 분석)

  • Lee, Jeongyeo;Min, Sung Ran;Jung, Jaeeun;Kim, HyeRan
    • Journal of Plant Biotechnology
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    • v.46 no.3
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    • pp.189-204
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    • 2019
  • In this study, we analyzed the changes in glucosinolate content and gene expression in TO1000DH3 and Early big seedling upon methyl jasmonate (MeJA) treatment. Analysis of glucosinolate contents after MeJA treatment at $200{\mu}M$ concentration showed that the total glucosinolate content increased by 1.3-1.5 fold in TO1000DH3 and 1.3-3.8 fold in Early big compared to those before treatment. Aliphatic glucosinolates, progoitrin and gluconapin, were detected only in TO1000DH3, and the changes in the content of neoglucobrassicin were the greatest at 48 hours after MeJA treatment in TO1000DH3 and Early big. The transcriptomic analysis showed that transcripts involved in stress or defense reactions, or those related to growth were specifically expressed in TO1000DH3, while transcripts related to nucleosides or ATP biosynthesis were specifically expressed in Early big. GO analysis on transcripts with more than two-fold change in expression upon MeJA treatment, corresponding to 12,020 transcripts in TO1000DH3 and 13,510 transcripts in Early big, showed that the expression of transcripts that react to stimulus and chemical increased in TO1000DH3 and Early big, while those related to single-organism and ribosome synthesis decreased. In particular, the expression increased for all transcripts related to indole glucosinolate biosynthesis, which is associated with increase in glucobrassicin and neoglucobrassicin contents. Upon MeJA treatment, the expression of AOP3 (Bo9g006220, Bo9g006240), TGG1 (Bo14804s010) increased only in TO1000DH3, while the expression of Dof1.1 (Bo5g008360), UGT74C1 (Bo4g177540), and GSL-OH (Bo4g173560, Bo4g173550, Bo4g173530) increased specifically in Early big.

Resveratrol Ameliorates NMDA-induced Mitochondrial Injury by Enhanced Expression of Heme Oxygenase-1 in HT-22 Neuronal Cells (NMDA를 처리한 HT-22 신경세포에서 미토콘드리아 손상을 완화하는 레스베라트롤의 보호 효과와 헴 산화효소-1의 역할)

  • Kang, Jae Hoon;Woo, Jae Suk
    • Journal of Life Science
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    • v.32 no.1
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    • pp.11-22
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    • 2022
  • N-methyl-D-aspartate (NMDA) receptors have received considerable attention regarding their involvement in glutamate-induced neuronal excitotoxicity. Resveratrol has been shown to exhibit neuroprotective effects against this kind of overactivation, but the underlying cellular mechanisms are not yet clearly understood. In this study, HT-22 neuronal cells were treated with NMDA in Mg2+-free buffer and subsequently used as an experimental model of glutamate excitotoxicity to elucidate the mechanisms of resveratrol-induced neuroprotection. We found that NMDA treatment causes a drop in MTT reduction ability, disrupts inside-negative transmembrane potential of mitochondria, depletes cellular ATP levels, and stimulates intracellular ROS production. Double fluorescence imaging studies demonstrated an increased formation of mitochondrial permeability transition (MPT) pores accompanied by apoptotic cell death, while cobalt protoporphyrin and bilirubin showed protective effects against NMDA-induced mitochondrial injury. On the other hand, zinc protoporphyrin IX significantly attenuated the protective effects of resveratrol which was itself shown to enhance heme oxygenase-1 (HO-1) mRNA and protein expression levels. In cells transfected with HO-1 small interfering RNA, resveratrol failed to suppress the NMDA-induced effects on MTT reduction ability and MPT pore formation. The present study suggests that resveratrol may prevent mitochondrial injury in NMDA- treated HT-22 cells and that enhanced expression of HO-1 is involved in the underlying cellular mechanism.

Regulatory Mechanism of Vascular Contractility by Extracellular $\textrm{K}^{+}$: Effect on Endothelium-Dependent Relaxation and Vascular Smooth Muscle Contractility (세포 외 $\textrm{K}^{+}$의한 혈관 수축신 조절 기전: 혈관평활근 수축성과 내피세포 의존성 이완에 미치는 영향)

  • 유지영;설근희;서석효;안재호
    • Journal of Chest Surgery
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    • v.37 no.3
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    • pp.210-219
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    • 2004
  • Extracellular $K^{+}$ concentration ([ $K^{+}$]$_{0}$ ) can be increased within several mM by the efflux of intracellular $K^{+}$. To investigate the effect of an increase in [ $K^{+}$]$_{0}$ on vascular contractility, we attempted to examine whether extracellular $K^{+}$ might modulate vascular contractility, endothelium-dependent relaxation (EDR) and intracellular $Ca^2$$^{+}$ concentration ([C $a^2$$^{+}$]$_{i}$ ) in endothelial cells (EC). We observed isometric contractions in rabbit carotid, superior mesenteric, basilar arteries and movse aorta. [C $a^2$$^{+}$]$_{i}$ was recorded by microfluorimeter using Fura-2/AM in EC. No change in contractility was recorded by the increase in [ $K^{+}$]$_{0}$ from 6 to 12 mM in conduit artery such as rabbit carotid artery. whereas resistant vessels, such as basilar and branches of superior mesenteric arteries (SMA), were relaxed by the increase. In basilar artery, the relaxation by the increase in [ $K^{+}$]$_{0}$ to from 1 to 3 mM was bigger than that by the increase from 6 to 12 mM. In contrast, in branches of SMA, the relaxation by the increase in [ $K^{+}$]$_{0}$ to from 6 to 12 mM is bigger than that by the increase from 1 to 3 mM. $Ba^2$$^{+}$ (30 $\mu$M) did not inhibit the relaxation by the increase in [ $K^{+}$]$_{0}$ from 1 to 3 mM but did inhibit the relaxation by the increase from 6 to 12 mM. In the mouse aorta without the endothelium or treated with $N^{G}$_nitro-L-arginine (30 $\mu$M), nitric oxide synthesis blocker, the increase in [ $K^{+}$]$_{0}$ from 6 to 12 mM did not change the magnitude of contraction induced either norepinephrine or prostaglandin $F_2$$_{\alpha}$. The increase in [ $K^{+}$]$_{0}$ up to 12 mM did not induce contraction of mouse aorta but the increase more than 12 mM induced contraction. In the mouse aorta, EDR was completely inhibited on increasing [ $K^{+}$]$_{0}$ from 6 to 12 mM. In cultured mouse aorta EC, [C $a^2$$^{+}$]$_{i}$ , was increased by acetylcholine or ATP application and the increased [C $a^2$$^{+}$]$_{i}$ , was reduced by the increase in [ $K^{+}$]$_{0}$ reversibly and concentration-dependently. In human umbilical vein EC, similar effect of extracellular $K^{+}$ was observed. Ouabain, a N $a^{+}$ - $K^{+}$ pump blocker, and N $i^2$$^{+}$, a N $a^{+}$ - $Ca^2$$^{+}$ exchanger blocker, reversed the inhibitory effect of extracellular $K^{+}$. In resistant arteries, the increase in [ $K^{+}$]$_{0}$ relaxes vascular smooth muscle and the underlying mechanisms differ according to the kinds of the arteries; $Ba^2$$^{+}$-insensitive mechanism in basilar artery and $Ba^2$$^{+}$ -sensitive one in branches of SMA. It also inhibits [C $a^2$$^{+}$]$_{i}$ , increase in EC and thereby EDR. The initial mechanism of the inhibition may be due to the activation of N $a^{+}$ - $K^{+}$pump. activation of N $a^{+}$ - $K^{+}$pump.p.p.p.