• Title/Summary/Keyword: Cultured cells

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Therapeutic Effects of Curdrania tricuspidata Leaf Extract on Osteoarthritis (골관절염 실험모델에서 꾸지뽕나무 추출물의 골관절염 억제효과 연구)

  • Nam, Da-Eun;Kim, Ok Kyung;Lee, Jeongmin
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.42 no.5
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    • pp.697-704
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    • 2013
  • The inhibitory effect of ethanol extracts from Curdrania tricuspidata leaves (CTL) on osteoarthritis was investigated in primary cultured rat cartilage cells and a monosodium-iodoacetate (MIA)-induced arthritis rat model. To identify the effects of CTL 80% ethanol extracts (CTL80) and CTL 10% ethanol extracts (CTL10) against $H_2O_2$ treatment in vitro, cell survival was measured by the MTT assay. Cell survival after $H_2O_2$ treatment increased with CTL80 and CTL10 close to normal up to $300{\mu}g/mL\;H_2O_2$. The mRNA expression of matrix metalloproteinases (MMPs) was determined MMP-7 and MMP-13 (known catabolic factors), were significantly inhibited by CTL 80 and CTL10; a $200{\mu}g/mL$ dose of CTL80 especially decreased MMP-13 expression. In vivo, osteoarthritis was induced by an intra-articular injection of MIA into the knee joints of rats, then CTL80 and CTL10 orally administered daily for 35 days. After the animals were sacrificed, histological evaluations of their knee joints revealed a reduction in polymorphonuclear cell infiltration and smooth synovial lining in the CTL80-500 group. Micro-CT analysis of hind paws from CTL80-500 and CTL10 showed a protection against osteophyte formation, soft tissue swelling, and bone resorption. In conclusion, CTL ethanol extracts are effective in ameliorating joint destruction and cartilage erosion in MIA-induced rats. CTL decreases and normalizes articular cartilage through preventing extracellular matrix degradation and chondrocyte injury, and could potentially serve as a therapeutic treatment for humans.

Growth Inhibition of Diatom, Chaetoceros calcitrans by Marine Bacteria, Shewanella sp. SR-14 - Effects of Marine Bacteria on the Changes of Fatty Acid Composition of Diatoms - (해양세균, Shewanella sp. SR-14에 의한 규조류 Chaetoceros calcitrans의 증식저해 - 해양세균이 규조류의 지방산 조성변화에 미치는 영향 -)

  • KIM Ji Hoe;YOON Ho Dong;PARK Hee Yun;LEE Hee Jung;CHANG Dong Suck
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.36 no.1
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    • pp.24-29
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    • 2003
  • In the previous reports, the authors isolated two strains of marine bacteria, Shewanella sp. SR-14, which has Chaetonros sp. growth inhibition activity, and Vibrio alginolyticus, that did not affect growth of the alga. In the present study, fatty acid compositions of diatoms, Chaetoceros calcitrans and Skeletonema costatum, and marine bacteria, Shewanella sp. SR-14 and V. alginolyticus, were analyzed. Changes of fatty acid composition in the diatoms grown with the marine bacteria were also determined. Major fatty acids of Sbewanella sp. SR-14 were 16:1n-7 $(29.4\%)$ and 16:0 $(19.2\%)$ during incubation in peptone broth at $20^{\circ}C$ for 3 days. The compositions of V. alginolyticus detected were 16:0 $(23.7\%),$ 16:1n-7 $(27.7\%)$ and 18:1n-7 $(21.0\%).$ C. calcitrans consisted of 16:1n-7 $(33.3\%),$ 16:0 $(27.1\%)$ and 14:0 $(12.1\%).$ S. costatum mainly contained 16:1n-7 $(28.9\%),$ 16:0 $(21.6\%)$ and 20:5 $(19.8\%).$ When halves of cell numbers of C. calcitrans were moribund cells by Shewanella sp. SR-14, the C. calcitrans and S. costatum simultaneously cultured with the bacteria were harvested by filtration with GE/D glass microfibre filter. In the fatty acid composition of both diatoms, saturated fatty acid contents in both diatoms grown with Shewanella sp. SR-14 were decreased, but unsaturated fatty acid contents were increased. The differences were greater in C. calcitrans than those in S. costatum. During the growth of diatoms with V. alginolyticus, C. calcitrans showed increase of saturated fatty acid contents and decrease of unsaturated fatty acid contents; however, S. costatum did not show sharp difference in fatty acid content. In this study, Shewanella sp. SR-14, which showed growth inhibition activity against C. calcitrans, influenced on the changes of fatty acid contents in the diatom. It was suggested that increased unsaturated fatty acid was synergistically activated algal growth inhibition activity of Shewanella sp. SR-14.

Physicochemical Properties and Physiological Activities of Rhus verniciflua Stem Bark Cultured with Fomitella fraxinea (장수버섯 균사체가 배양된 옻피의 이화학적 특성 및 생리활성)

  • Choi, Han-Seok;Kim, Bo-Hyun;Yeo, Soo-Hwan;Jeong, Seok-Tae;Choi, Ji-Ho;Park, Hyo-Suk;Kim, Myung-Kon
    • The Korean Journal of Mycology
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    • v.38 no.2
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    • pp.172-178
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    • 2010
  • The contents of proximate composition, free amino acids and phenolic acids in the Fomitella fraxinea cultivated-Rhus verniciflua stem bark(FRVSB), and its adipogenesis effect were investigated. The proximate composition(%) of FRVSB was as follows: moisture(7.64), ash(6.30), crude fat(3.86), crude protein(3.59) and sugar(not detected); while Rhus verniciflua stem bark(RVSB) contained 1.64, 8.09, 7.28, 6.48 and 5.39, respectively. The total free amino acids concentration was 97.41 mg% in FRVSB and 71.91 mg% in RVSB. Phosphoserine(55.06 mg%), ammonia(17.84mg%) and aspartic acid(6.05mg%) were predominant amino acids. The content of total phenolic acids was 422.89 ppm in ethanol extract and 283.86 ppm in water extract, with syringic and gallic acid as the main component. The FRVSB extracts showed a potent free radical scavenging activity for DPPH(2,2-diphenyl-1-picrylhydrazyl hydrate) with $IC_{50}$ of $28.54\;{\mu}g$(EtOH) and $54.70\;{\mu}g$(water), respectively, whereas $IC_{50}$ value of gallic acid was $1.84\;{\mu}g$. The protective effect of both ethanol and water extract the extracts against UV-induced oxidative stress in NIH3T3 was observed. The water extracts of FRVSB may promote adipogenesis in 3T3-L1 cells.

Anti-inflammatory Effect and Antioxidative Activities of Ingredients used in Bibimbab (비빔밥 재료의 항염증 및 항산화 효과)

  • Ko, Yu-Jin;Seol, Hui-Gyeong;Lee, Gyeong-Ran;Jeong, Gye-Im;Ryu, Chung-Ho
    • Journal of Life Science
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    • v.23 no.2
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    • pp.213-221
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    • 2013
  • Bibimbab (mixed rice) is a traditional Korean one-dish meal. This study was carried out to investigate the anti-inflammatory and antioxidative effects of raw and seasoned ingredients used in Bibimbab (Cucurbita moschata P., Platycodon grandiflorum A., Vigna radiata L., Porphyra yezonensis udea, Allium ampeloprasum L., Pterdium aguilinum, Raphanus sativus). Human mast cells (HMC-1) were pretreated with 70% ethanol extracts of Bibimbab and further cultured for an appropriate time after the addition of phorbol 12-myristate 13-acetate (PMA) and calcium ionophore A23187. Cell viability was determined by an MTT assay. None of the ingredients showed cytotoxic effects at a concentration of 1.0 ${\mu}g/ml$. Anti-inflammatory effects were analyzed at 0.01, 0.1, and 1.0 mg/ml concentrations of the raw, seasoned ingredients of PMA. A23187 stimulated HMC-1. Among the various ingredients, seasoned A. ampeloprasum L. extract showed the highest inhibition of TNF-${\alpha}$ and IL-6 secretion (90% and 93%, respectively) at a concentration of 1.0 mg/ml. The R. sativus extract showed the highest inhibition (85%) of IL-8 secretion. DPPH analysis of the antioxidation properties of the ingredients showed that raw and seasoned A. ampeloprasum extracts exhibited the highest DPPH free radical scavenging activity (67.50 and 73.65%, respectively). These results suggest that seasoned ingredients used in Bibimbab have lower anti-inflammatory effects in relation to TNF-${\alpha}$ and IL-6 secretion than raw ingredients in PMA- and A23187-treated HMC-1. In addition, the seasoned ingredients showed a tendency to increase antioxidative activity. Therefore, the ingredients used in Bibimbab have potential as anti-inflammatory and antioxidation agents.

The Effect of Kaempferol, guercetin on Hyaluronan-Synthesis Stimulation in Human Keratinocytes (HaCaT) (인체 피부 세포주 (HaCaT)에서 Kaempferol, Quercetin의 Hyaluronan 합성 촉진 효과에 대한 연구)

  • Kim, Seung-Hun;Nam, Gae-Won;Kang, Byung-Young;Lee, Hae-Kwang;Moon, Seong-Joon;Chang, Ih-Seop
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.31 no.1 s.49
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    • pp.97-102
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    • 2005
  • One of the key molecules involved in skin moisture is hyaluronan (hyaluronic acid, HA) with its associated water of hydration. The predominant component of the ECM (extracellular matrix) of skin is HA. It Is the primordial and the simplest of the GAGs (glycosaminoglycans), a water-sorbed macromolecule In extracellular matrix, Included between the vital cells of epidermis. In the skin, HA was previously thought to derive extlusively from dermis. But, recent studies revealed that HA could be synthesized in epidermis. Flavonoids are polyphenolic compounds that is found mainly in foods of plant origin. Kaempferol was known to increase glutathione synthesis in human keratinocyte. And quercetin blocked PPAR-meidated keratinocyte differentiation as lipoxygenase inhibitors. In this study, we sought to evaluate the effect of flavonid, kaempferol and quercetin on production HA in keratinocyte. We examined the changes of three human hyaluronan synthase genes (HASI, HAS2, HAS3) expression by semi-quantitative RT-PCR when kaempferol or quercetin was added to cultured human keratinocytes. We found that these flavonoids slightly upregulated HAS2, HAS3 mRNA after 24 h. And we investigated the effect on HA production by ELISA. When we evaluated the level of HA in culture medium after 24 h incubation. We found enhanced accumulation of HA in the culture medium. Although the effects of above flavonoids are less than retinoic acid, the data indicate that kaempferol, quercetin can dose-dependently increase the level of HA in epidermis cell line. It suggested that flavonoid, kaempferol, and quercetin increased production of HA in skin and it helped to elevate skin moisture and improve facial wrinkle.

Production of Poly(3-hydroxybutyrate) Using Waste Frying Oil (Waste frying oil를 사용한 Poly(3-Hydroxybutyrate) 생합성)

  • Kim, Tae-Gyeong;Lee, Woosung;Gang, Seongho;Kim, Jong-Sik;Chung, Chung-Wook
    • Journal of Life Science
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    • v.29 no.1
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    • pp.76-83
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    • 2019
  • In this study, the optimal growth and poly(3-hydroxybutyrate) (PHB) biosynthesis of Pseudomonas sp. EML2 were established using waste frying oil (WFO) as a cheap carbon source. The fatty acid composition of WFO and fresh frying oil (FFO) were analyzed by gas chromatography. The unsaturated and saturated fatty acid contents of the FFO were 82.6% and 14.9%, respectively. These contents changed in the WFO. The compositional change in the unsaturated fatty acid content in the WFO was due to a change in its chemical and physical properties resulting from heating, an oxidation reaction, and hydrolysis. The maximum dry cell weight (DCW) and PHB yield (g/l) of the isolated strain Pseudomonas sp. EML2 were confirmed under the following culture conditions: 30 g/l of WFO, 0.5 gl of $NH_4Cl$, pH 7, and $20^{\circ}C$. Based on this, the growth and PHB yield of Pseudomonas sp. EML2 were confirmed by 3 l jar fermentation. After the cells were cultured in 30 g/l of WFO for 96 h, the DCW, PHB content, and PHB yield of Pseudomonas sp. EML2 were 3.6 g/l, 73 wt%, and 2.6 g/l, respectively. Similar results were obtained using 30 g/l of FFO as a carbon source control. Using the FFO, the DCW, PHB content, and PHB yield were 3.4 g/l, 70 wt%, and 2.4 g/l, respectively. Pseudomonas sp. EML2 and WFO may be a new candidate and substrate, respectively, for industrial production of PHB.

Enhancement of Bioactive Compounds in Mugwort Grown under Hydroponic System by Sucrose Supply in a Nutrient Solution (양액 내 자당 처리에 의한 수경재배 쑥의 생리활성물질 증진)

  • Moon-Sun Yeom;Jun-Soo Lee;Myung-Min Oh
    • Journal of Bio-Environment Control
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    • v.32 no.1
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    • pp.23-33
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    • 2023
  • Sucrose (suc) is a disaccharide that consists of glucose (glu) and fructose (fru). It is a carbohydrate source that acts as a nutrient molecule and a molecular signal that regulates gene expression and alters metabolites. This study aimed to evaluate whether suc-specific signaling induces an increase in bioactive compounds by exogenous suc absorption via roots or whether other factors, such as osmotic stress or biotic stress, are involved. To compare the osmotic stress induced by suc treatment, 4-week-old cultured mugwort plants were subjected to Hoagland nutrient solution with 10 mM, 30 mM, and 50 mM of suc or mannitol (man) for 3 days. Shoot fresh weight in suc and man treatments was not significantly different from the control. Both man and suc treatments increased the content of bioactive compounds in mugwort, but they displayed different enhancement patterns compared to the suc treatments. Mugwort extract treated with suc 50 mM effectively protected HepG2 liver cells damaged by ethanol and t-BHP. To compare the biotic stress induced by suc treatment, 3-week-old mugwort plants were subjected to microorganism and/or suc 30 mM with Hoagland nutrient solution. Microorganisms and/or suc 30 mM treatments showed no difference about the shoot fresh weight. However, sugar content in mugwort treated with suc 30 mM and microorganism with suc 30 mM treatment was significantly higher than that of the control. Suc 30 mM and microorganism with suc 30 mM were effective in enhancing bioactive compounds than microorganism treatment. These results suggest that mugwort plants can absorb exogenous suc via roots and the enhancement of bioactive compounds by suc treatment may result not from osmotic stress or biotic stress because of microorganism, but by suc-specific signaling.

Effects of oxypeucedanin hydrate isolated from Angelica dahurica on myoblast differentiation in association with mitochondrial function (백지에서 추출한 oxypeucedanin hydrate의 미토콘드리아 기능 관련 근생성 효과)

  • Eun-Ju Song;Ji-Won Heo;Jee Hee Jang;Yoon-Ju Kwon;Yun Hee Jeong;Min Jung Kim;Sung-Eun Kim
    • Journal of Nutrition and Health
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    • v.57 no.1
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    • pp.53-64
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    • 2024
  • Purpose: Mitochondria play a crucial role in preserving skeletal muscle mass, and damage to mitochondria leads to muscle mass loss. This study investigated the effects of oxypeucedanin hydrate, a furanocoumarin isolated from Angelica dahurica radix, on myogenesis and mitochondrial function in vitro and in zebrafish models. Methods: C2C12 myotubes cultured in media containing 0.1, 1, 10, or 100 ng/mL oxypeucedanin hydrate were immunostained with myosin heavy chain (MHC), and then multinucleated MHC-positive cells were counted. The expressions of markers related to muscle differentiation, muscle protein degradation, and mitochondrial function were determined by quantitative reverse transcription polymerase chain reaction. To investigate the effects of oxypeucedanin hydrate on mitochondrial dysfunction, Tg(Xla.Eef1a1:mito-EGFP) zebrafish embryos were treated with 5-fluorouracil, leucovorin, and irinotecan (FOLFIRI) with or without oxypeucedanin hydrate and analyzed for mito-EGFP intensity and mitochondrial length. Results: Oxypeucedanin hydrate significantly increased MHC-positive multinucleated myotubes (≥ 3 nuclei) and increased the expression of the myogenic marker myosin heavy chain 4. However, it decreased the expressions of muscle-specific RING finger protein 1 and muscle atrophy f-box (markers of muscle protein degradation). Furthermore, oxypeucedanin hydrate enhanced the expressions of markers of mitochondrial biogenesis (peroxisome proliferator-activated receptor-gamma coactivator 1 alpha, transcription factor a mitochondrial, succinate dehydrogenase complex flavoprotein subunit A, and cytochrome c oxidase subunit 1) and mitochondrial fusion (optic atrophy 1). However, it reduced the expression of dynamin-related protein 1 (a mitochondrial fission regulator). Consistently, oxypeucedanin hydrate reduced FOLFIRI-induced mitochondrial dysfunction in the skeletal muscles of zebrafish embryos. Conclusion: The study indicates that oxypeucedanin hydrate promotes myogenesis by improving mitochondrial function, and thus, suggests oxypeucedanin hydrate has potential use as a nutritional supplement that improves muscle mass and function.

Larvae Growth and Biochemical Composition Change of the Pacific Oyster Crassostra gigas, Larvae during Artificial Seed Production (참굴 Crassostrea gigas 인공종묘생산 시 유생의 성장과 체성분 변화)

  • Hur, Young-Baek;Min, Kwang-Sik;Kim, Tae-Eic;Lee, Seung-Ju;Hur, Sung-Bum
    • Journal of Aquaculture
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    • v.21 no.4
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    • pp.203-212
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    • 2008
  • A nutritional demand of oyster, Crassostrea gigas larva as part of research for improving of utilization of microalgae being used for the artificial oyster seed production. The change of body growth and biochemical compositions of larvae were investigated during larvae rearing in hatchery. The larvae were cultured in 60 M/T tank and fed mixture 6 different phytoplankton species, Isochrysis galbana (30%), Cheatoceros gracilis (20%), Pavlova lutheri (20%), Phaeodactylum triconutum (10%), Nannochryis oculata (10%) and Tetraselmis tetrathele (10%). The initial feeding amount was $0.3{\times}10^4cells/mL$ at three times a day to D-shaped larva and the feeding amount had been increased 30% gradually every two day since the larvae were raising. The larvae were developed from D shape to pediveliger stage for 12 days. The daily growth of shell length and hight were $5.8{\sim}30.8\;{\mu}m$ and $8.7{\sim}31.4\;{\mu}m$, respectively and weight gains were changed from D shape to pediveliger as follow: wet weight was $0.52{\sim}15.0\;{\mu}g/larva$, dry weight was $0.2{\sim}6.5\;{\mu}g/larva$, and ash free dry weight was $0.1{\sim}8.5\;{\mu}g/larva$. The larvae growth pattern shown a logarithmic phase from D shape to umbone stage but after that stage shown a exponential growth aspect. The crude protein, crude lipid and nitrogen free extract (NFE) of larvae during rearing periods were analyzed as $6.1{\sim}10.6%$, $0.6{\sim}1.1%$ and 1.0-2.7%, respectively. And the total amino acid contents of the larvae during rearing periods were in order as glutamic acid $1.26{\sim}2.24%$, aspartic acid $0.97{\sim}1.70%$, and methionine $0.12{\sim}0.33%$. Of the total fatty acid in the analyzed larvae, the saturated fatty acid (SSAFA) was decreased from 54.3% (D shaped larvae) to 17.1 % (pediveliger) as larvae development but the total mono-unsaturated fatty acid (${\Sigma}MOFA$) and Poly-unsaturated fatty acid (${\Sigma}PUFA$) were increased from 29.9% and 7.8% to 40.6% and 45.6%, respectively. By the way the each fatty acid of the larvae were composed of palmitic acid $9.89{\sim}36.95%$, oleic acid $12.17{\sim}32.29%$, linoleic acid $1.96{\sim}33.55%$, EPA $2.17{\sim}11.58%$ and DHA $1.95{\sim}4.51%$. As a result of this study, the larvae of oyster were demanded a various nutrients for healthy growth and the feeding control, expecially after umbone stage larvae are a rapidly growing time, is very important for success of artificial seed production.

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.