• Title/Summary/Keyword: L-NNA

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Nitric Oxide-mediated Relaxation by High $K^+$ in Human Gastric Longitudinal Smooth Muscle

  • Kim, Young-Chul;Choi, Woong;Yun, Hyo-Young;Sung, Ro-Hyun;Yoo, Ra-Young;Park, Seon-Mee;Yun, Sei-Jin;Kim, Mi-Jung;Song, Young-Jin;Xu, Wen-Xie;Lee, Sang-Jin
    • The Korean Journal of Physiology and Pharmacology
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    • v.15 no.6
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    • pp.405-413
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    • 2011
  • This study was designed to elucidate high-$K^+$ induced response of circular and longitudinal smooth muscle from human gastric corpus using isometric contraction. Contraction from circular and longitudinal muscle stripes of gastric corpus greater curvature and lesser curvature were compared. Circular smooth muscle from corpus greater curvature showed high $K^+$ (50 mM)-induced tonic contraction. On the contrary, however, longitudinal smooth muscle strips showed high $K^+$ (50 mM)-induced sustained relaxation. To find out the reason for the discrepancy we tested several relaxation mechanisms. Protein kinase blockers like KT5720, PKA inhibitor, and KT5823, PKG inhibitor, did not affect high $K^+$-induced relaxation. $K^+$ channel blockers like tetraethylammonium (TEA), apamin (APA), glibenclamide (Glib) and barium ($Ba^{2+}$) also had no effect. However, N(G)-nitro-L-arginine (L-NNA) and 1H-(1,2,4) oxadiazolo (4,3-A) quinoxalin-1-one (ODQ), an inhibitor of soluble guanylate cyclase (sGC) and 4-AP (4-aminopyridine), voltage-dependent $K^+$ channel (KV) blocker, inhibited high $K^+$ -induced relaxation, hence reversing to tonic contraction. High $K^+$-induced relaxation was observed in gastric corpus of human stomach, but only in the longitudinal muscles from greater curvature not lesser curvature. L-NNA, ODQ and KV channel blocker sensitive high $K^+$-induced relaxation in longitudinal muscle of higher portion of corpus was also observed. These results suggest that longitudinal smooth muscle from greater curvature of gastric corpus produced high $K^+$-induced relaxation which was activated by NO/sGC pathway and by $K_V$ channel dependent mechanism.

Nitric Oxide Synthase Mediates Carbon Monoxide-Induced Stimulation of L-type Calcium Currents in Human Jejunal Smooth Muscle Cells

  • Lim, In-Ja;Yun, Ji-Hyun;Kim, Seung-Tae;Myung, Soon-Chul;Kim, Tae-Ho;Bang, Hyo-Weon
    • The Korean Journal of Physiology and Pharmacology
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    • v.8 no.3
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    • pp.161-165
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    • 2004
  • Exogenous carbon monoxide (0.2%) increases L-type calcium $(Ca^{2+})$ current in human jejunal circular smooth muscle cells. The stimulatory effect of carbon monoxide (CO) on L-type $Ca^{2+}$ current is inhibited by pre-application of L-NNA, a classical competitive inhibitor of nitric oxide synthase (NOS) with no significant isoform selectivity (Lim, 2003). In the present study, we investigated which isoform of NOS affected CO induced stimulation of L-type $Ca^{2+}$ current in human jejunal circular smooth muscle cells. Cells were voltage clamped by whole-cell mode patch clamp technique, and membrane currents were recorded with 10 mM barium as the charge carrier. Before the addition of CO, cells were pretreated with each inhibitor of three NOS isoforms for 15 minutes. CO-stimulating effect on L-type $Ca^{2+}$ current was partially blocked by N-(3-(Amino-methyl) benzyl) acetamidine 2HCl (1400W, an iNOS inhibitor). On the other hand, 3-bromo-7-nitroindazole (BNI, a nNOS inhibitor) or $N^5-(1-Iminoethyl)-L-ornithine$ dihydrochloride (L-NIO, an eNOS inhibitor) completely blocked the CO effect. These data suggest that low dose of exogenous CO may stimulate all NOS isoforms to increase L-type $Ca^{2+}$ channel through nitric oxide (NO) pathway in human jejunal circular smooth muscle cells.

Effect of Kamichungbieum on the Isolated Rat Aorta (가미청비음이 흰쥐의 적출 동맥에 미치는 영향)

  • Eun Jae Soon;Lee Dong Hee;Han Jong Hyun
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.18 no.4
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    • pp.1107-1110
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    • 2004
  • The purpose of this research was to investigate the effects of supercritical fluid extract of Kamichungbieum (SFE) on the contraction of isolated rat aorta. The contractile force of rat aorta was measured with force displacement transducer under 1.5g loading tension. The contraction of aorta induced by phenylephrine 0.1 μM was inhibited by SFE. The aorta relaxed by SFE was inhibited by the pretreatment of L-NNA, ODQ or indomethacin, respectively. These results indicate that SFE induce the relaxation of isolated aorta via activation of nitric oxide, cAMP and cyclooxygenase in epithelium cells.

Effect of Yongyanggaktang(YYG) on Regional Cerebral Blood Flow in Rats (영양각탕(羚羊角湯)이 뇌혈류량(腦血流量)에 미치는 영향(影響))

  • Kang Sung-Hyun;Yun Young-Gab
    • Herbal Formula Science
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    • v.10 no.1
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    • pp.105-112
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    • 2002
  • Yongyanggaktang has been used in oriental medicine as a therapeutic agent for cerebral disease and eclampsia for many years. The purpose of this study was to determine the effect of Yongyanggaktang and its exception of Cornu Saigae Tataricae(羚羊角) in YYG about regional cerebral blood flow(rCBF) in rats. 1. rCBF was increased by Yongyanggaktang in dose dependently. 2. rCBF was increased by exception of Cornu Saigae Tataricae(羚羊角) in YYG at high dosages. 3. rCBF was decreased by Yongyanggaktang pretreated propranolol. 4. L-NNA did not affect rCBF. 5.ODQ did not affect rCBF. Above the results. Yongyanggaktang relates to the symphathetic action.

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Impact of Physiological Stresses on Nitric Oxide Formation by Green Alga, Scenedesmus obliquus

  • Mallick, Nirupama;Mohn, Friedrich-Helmuth;Rai, Lalchand;Soeder, Carl-J.
    • Journal of Microbiology and Biotechnology
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    • v.10 no.3
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    • pp.300-306
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    • 2000
  • The rate of apparent nitric oxide (NO) release, as measured in the exhaust gas of green alga, Scenedesmus obliquus, depended on the light intensity and pH. It doubled after lowering the temperature from $25^{\circ}C{\;}to{\;}15^{\circ}C$ and strongly decreased from $35^{\circ}C{\;}to{\;}42^{\circ}C$. The Scenedesmus cells, deficient in nitrogen or phosphorus, demonstrated a significant increase in NO production following their transfer to nitrate- and phosphate-rich media. The addition of herbicides (DCMU and glyphosate) or toxic concentrations of $Cu^{2+}{\;}or{\;}Fe^{3+}$ produced strong NO peaks, resembling those that occurred after sudden darkening. An increase in the $Ni^{2+}$ concentration to 20 ppm resulted in a gradual increase of NO release from the initial ~1.5 ppbv to>20 ppbv, whereas $Cd^{2+}$ instantaneously suppressed the NO by the cultures of Scenedesmus was not altered by L-NNA, an inhibitor of nitric oxide synthase (NOS), or by its substrate, L-arginine. This seems to exclude the role of NOS in the NO formation under study. Accordingly, it can be assumed that the rate of NO formation is mainly a function of dynamic nitrite pool sizes and environmental factors significantly affect the NO production in algae.

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Seed Pelletizing of Salvia splendens and Calendula officinalis for the Greening and Re-vegetating (녹화와 식생조성을 위한 샐비어와 금잔화 종자의 펠렛처리)

  • Kim, Seung-Hyun;Lee, Byoung-Ryong;Choi, Byoung-Kon
    • Journal of the Korean Society of Environmental Restoration Technology
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    • v.10 no.5
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    • pp.67-75
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    • 2007
  • The purpose of this study was to pelletize to calendula and salvia seeds with peat moss as basic material for the efficient greening and vegetating of slopes and damaged areas. Also the pelletizing was compressed by spherical types that mixed basic fertilizer of N.;300 mg/l, P.;200 mg/l, K.;400 mg/g and plant growth regulator of A and NAA each 300PPM. Soil and soil surface seeding methods were researched to find the growing state of germination percent, germination date, germination force, length of leaf, number of leaf, width of leaf, length of plant, and etc. Comparing with controlled pelletizing, peat moss and +GA pelletizing treatments resulted in higher from two to three times as following growing states : length of leaves, number of leaves, width of leaves, length of plants, length of roots, fresh weights, and ratio of germination. Especially the two treatments above showed four more times effects than the +NNA treatment. Also their germinating date germinating force were earlier and stronger. The surface seeding method was superior to soil seeding.

Effects of GwakHyangJungGiSan on the Arterial Contraction in Rabbit (곽향정기산(藿香正氣散)이 가토(家兎)의 수축혈관에 마치는 영향(影響))

  • Sun Jung-Ki;Kim Ho-Hyun;Nam Chang-Gyu;Koo Chang-Mo
    • The Journal of Internal Korean Medicine
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    • v.24 no.2
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    • pp.260-268
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    • 2003
  • Object : This study was undertaken to define the mechanism of GwakHyangJungGiSan-induced relaxation in rabbit common carotid artery contracted by agonists. Method : In order to investigate the effect of GwakHyangJungGiSan on rabbit's contracted vascular ring detached from common carotid artery, vascular ring intact or damaged endothelium was used for the experiment using organ bath. To analyze the mechanism of GwakHyangJungGiSan-induced relaxation, GwakHyangJungGiSan extract was infused into contracted vascular ring which had been pretreated by pretreatment of indomethacin(IM), tetraethylammonium chloride(TEA), $N{\omega}-nitro-L-arginine(L-NNA)$. Result : GwakHyangJungGiSan blocks an inflow of $Ca^{2+}$ and relaxes vascular ring by the action of Nitric oxide from endothelium. Consequently when GwakHyangJungGiSan is prescribed, a rise in blood pressure by the resistance of peripheral vessel may be controlled to some extent and so it is anticipated that hypertension, a disorder of blood flow from the vascular contraction and vascular disease will be treated well.

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Vasorelaxant Activities of Aqueous Extracts from Twenty Medicinal Plants Used in Oriental Medicines in Isolated Rat Aorta

  • Kim, Eun-Young;Lee, Kyung-Ok;Kim, Dong-Il;Rhyu, Mee-Ra
    • Preventive Nutrition and Food Science
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    • v.15 no.3
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    • pp.189-195
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    • 2010
  • Water extracts from 20 medicinal plants, traditionally used for postmenopausal symptoms in Korea, were examined for their vasorelaxant activity in isolated rat thoracic aorta rings precontracted with norepinephrine (NE). Among the 20 medicinal plants, Cornus officinalis (CoEx, 0.3 mg/mL), Schisandra chinensis (ScEx, 0.3 mg/mL), Erythrina variegate (EvEx, 0.3 mg/mL), and Epimedium koreanum (EkEx, 0.3 mg/mL) showed rapid relaxation of endothelium-intact aorta ($69\pm4%$, $40\pm3%$, $25\pm2%$, and $23\pm3%$ of active tone induced by NE, respectively). In contrast, the extracts of Erythrina variegata (EvEx), Angelica gigas (AgEx), Pueraria thunbergiana (PtEx), and EkEx lead to gradual (i.e., long-term) relaxation to baseline in endothelium-intact vessels. The time to complete relaxation was 20~40 min. These 6 plant extracts were selected for the investigation of possible underlying mechanisms. The CoEx-, ScEx-, or EkEx-induced rapid relaxations were virtually abolished by endothelium denudation, and were significantly inhibited by pretreatment with nitric oxide (NO) synthase inhibitor $N^G$-nitro-L-arginine (L-NNA, 10 ${\mu}M$), indicating that increased formation of NO might contribute to the endothelium-mediated relaxation. In long-term responses, the endothelium denudation did not affect PtEx-induced relaxation, whereas it delayed responses by EvEx and AgEx, and significantly inhibited the effect of EkEx. Among EvEx, AgEx, and PtEx, EvEx attenuated the $CaCl_2$-induced vasoconstriction in high-potassium depolarized medium, implying that EvEx is involved in inhibition of the extracellular calcium influx to smooth muscle through voltage dependent calcium channels. These results provide the scientific rationale for the interrelationships between the use of 20 medicinal plants and their effects on cardiovascular health in estrogen deficient conditions.

Effects of Medicinal Plant Extract on the Change of Cerebral Hemodynamic in Rats (약용식물 추출물이 흰쥐의 뇌혈류학적 변화에 미치는 영향)

  • Park, Sung-Jin;Hahm, Tae-Shik;Kim, Cheun-An
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.39 no.4
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    • pp.506-510
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    • 2010
  • As an attempt to develop new functional health beverage by using medicinal herb, we investigated the effect of medicinal plant extract (MPE) on mean arterial blood pressure (MABP) and regional cerebral blood flow (rCBF) of rats. The changes of MABP and rCBF were determined by LDF methods. LDF allows for real time, noninvasive, continuous recordings of local CBF. MABP in MPE treated rats showed significant change of MPE 1.0 and 10.0 mg/kg. MPE i.v. administration showed significant increase of rCBF in a dose-dependent manner. Propranolol pretreated MABP showed significant change in the increase of MPE. rCBF of propranolol pretreated rats showed significant change from the i.v. injection concentration of 1.0 and 10.0 mg/kg. The ischemia/reperfusion induced oxidative stress may have contributed to cerebral damage in rats, and the present study provides clear evidence for the beneficial effect of MPE on ischemia induced brain injury. Also, the action mechanism in elevation effect of MPE on rCBF might be concerned with the role of $\beta$-adrenoceptor. The exact component and mechanism remains for the future study.

Purification and Characterization

  • Nam, Suk-Woo;Seo, Dong-Wan;Sung, Dae-Seok;Han, Jeung-Whan;Hong, Sung-Youl;Lee, Hyang-Woo
    • Archives of Pharmacal Research
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    • v.21 no.2
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    • pp.128-134
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    • 1998
  • Nitric oxide synthase, NOS (EC.1.14.13.39), was purified from bovine pancreas over 5,500-fold with a 7.6% yield using 30% ammonium sulfate precipitation, and $2^1$,$5^1$-ADP-agarose and calmodulin-agarose affinity chromatography. The purified bovine pancreatic NOS (bpNOS) showed a single band on SDS-PAGE corresponding to an apparent molecular mass of 160 kDa, whereas it was 320 kDa on non-denaturating gel-filtration. This indicated a homodimeric nature of the enzyme. The specific activity of the purified bpNOS was 31.67 nmol L-citrulline fored/mtn/mg protein and an apparent $K\textrm{m}$ for L-arginine was 15.72 $\mu\textrm{M}$, The enzyme activity was dependent on $Ca^{2+}$ and calmodulin, and to a lesser extent on NADPH, FAD and FMN. $H_4B$ was not required as a cofactor for the activity. In an inhibition experiment with L-arginine analogues, $N^G$-nitro-L-arginine (NNA) had the most potent inhibitory effect on bpNOS, and $N^{G}$, $N^{G1}$-dimethyl-L-arginine (symmetric; sDMA) did not have any inhibitory effect. Immunohistochemical analysis of the bovine pancreas using brain type NOS antibody (anti-bNOS antibody) revealed that acinar cells showed strong immunoreactivity against the antibody.

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