• Title/Summary/Keyword: L-Arginine

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Mode of Action of Streptomycin Resistance in the Citrus Canker Pathogen (Xanthomonas smithii subsp. citri) in Jeju Island

  • Hyun, Jae-Wook;Kim, Hyo-Jung;Yi, Pyoung-Ho;Hwang, Rok-Yeon;Park, Eun-Woo
    • The Plant Pathology Journal
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    • v.28 no.2
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    • pp.207-211
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    • 2012
  • It has been known that streptomycin resistance in bacteria can occur as a results of chromosomal mutation or through gene acquisition or both. Chromosomal mutations for resistances are point mutations in the rpsL gene, which alter ribosomal protein S12. Acquired resistance has occurred when an $Sm^R$ plasmid carrying transposon Tn5393 with tandem strA-strB gene is transferred by conjugation. A total of 686 isolates of Xanthomonas smithii subsp. citri causal agent of citrus canker disease were collected from 26 citrus orchards in Jeju Island in 2003 and 2004 seasons. Forty-nine of 111 isolates from streptomycin non-sprayed orchards in 2003 season were resistant to streptomycin. Of 107 isolates from orchards sprayed one time with streptomycin, 58 isolates were resistant, and 166 of 221 isolates from orchards sprayed two times with streptomycin were resistant. In 12 orchards sprayed three or more times with streptomycin, 219 of 247 isolates were resistant to streptomycin. Twenty-five isolates of X. smithii subsp. citri were surveyed to identify the mechanisms of streptomycin resistance in this study. Twenty-one of these 25 isolates were resistant to streptomycin, and it was proven by PCR assay that 18 of the 21 streptomycin resistant isolates have the strB gene. In sixteen of the 21 streptomycin resistant isolates, it was occurred a point mutation altered codon lysine (AAG)-41 of rpsL gene to arginine (AGG). The streptomycin-sensitive isolates easily acquired the resistance by mixed culture with resistant isolates. The strB gene was amplified from the isolates that acquired the resistance by mixed culture, and one isolate of them was also point-mutated in codon 41 of rpsL gene to be resistant. In this study, most of the streptomycin-resistant isolates of X. smithii sub sp. citri in Jeju island expressed the resistance by both chromosomal point mutation and gene acquisition, and the resistance was easily acquired through conjugation by culture mixed with streptomycin resistant and sensitive strains.

Effect of Extracting Conditions on Chemical Compositions of Korean Mountain Ginseng Extract (추출조건이 장뇌삼추출물의 화학성분 조성에 미치는 영향)

  • Kim, Jun-Han;Kim, Jong-Kuk
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.34 no.6
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    • pp.862-868
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    • 2005
  • Korean mountain ginseng roots were freeze-dried at $-70^{\circ}C$ and extracted by different extracting solution conditions to investigate chemical compositions of extracts. The soluble solid content of the extract from $7.04\~13.45\%\;and\;50\%$ EtOH and MeOH extracts were higher than those of other extracts. $100\%\;water\;and\;90\%$ EtOH extracts gave the highest Brix with $19.98\%$\;and\;19.65\%$, respectively. pH of the extracts were ranged from $5.82\~6.60$. Browning color at 470 nm of the extract were high value in 50$\%$ EtOH extract. In case of Hunter's color value, L value of extract was higher in $100\%$ water extract (21.28) than EtOH extract $(17.18\~21.02)$, a and b values of extract were the highest in $100\%$ water (-0.12) and $90\%$ MeOH extract (1.34). The contents of free sugars in the EtOH extract were increased with the ethanol concentration. Sucrose contents of $90\%$ EtOH and MeOH extracts were 6,159 mg/100 g and 5,238 mg/100 g. Major organic acids of the extract were citric and malic acids. Major free amino acids of the extract were L-arginine, L-proline, $\gamma$ -amino-n-butyric acid, alanine and aspartic acid. The highest ginsenoside content was shown to be about $10.50\%\;in\;90\%$ MeOH extract. Major minerals of extract were P, K, Na, Mg and Ca.

Analysis of Amino Acids, Fatty Acids, and Antioxidant Activities of Prunus yedoensis Matsum. Bark Extracts (왕벚나무 수피 추출물의 아미노산, 지방산 분석 및 항산화 활성)

  • Sung-Hwan Park;Ye-Eun Choi;Jung-Mo Yang;Chae-Won Jeong;Hyun-Duck Jo;Ju-Hyun Cho
    • Journal of Food Hygiene and Safety
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    • v.39 no.3
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    • pp.288-298
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    • 2024
  • This study aimed to investigate the amino acid content, fatty acid composition, total flavonoid and phenol contents, and antioxidant activities of Prunus yedoensis Matsum. bark extracts using various extraction solvents. Through amino acid analysis, 13 amino acids were detected in extracts obtained using hot water, 30% ethanol, and 70% ethanol. The major amino acids were identified as aspartic acid, arginine, and proline, and the total amino acid content was 0.17%, 0.16%, and 0.09%, respectively. Fatty acid analysis showed a saturated fatty acid (SFA) ratio of 62.7-66.7% in extracts obtained using hot water, 30% ethanol, and 70% ethanol, with the primary fatty acid identified as palmitic acid. The total flavonoid and polyphenol contents were 727.70-769.87 mg quercetin equivalent (QE)/ g and 309.24-348.09 mg gallic acid equivalent (GAE)/g, respectively, in extracts obtained using hot water, 30% ethanol, and 70% ethanol extracts. Measurements of antioxidant activity confirmed that extracts obtained using hot water, 30% ethanol, and 70% ethanol extracts increased the antioxidant effect in 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging activity, 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) radical scavenging activity, and ferric reducing antioxidant power (FRAP) in a concentration-dependent manner. A high correlation was observed between the total flavonoid content, total polyphenol content, and antioxidant activities of the extracts. This study provides data and novel insights for the development of functional food materials using P. yedoensis Matsum. bark extracts.

Studies on the Chemical Constituents of the New Zealand Deer Velvet Antler Cervus elaphus var. scoticus-(I)

  • Lee, Nam Kyung;Shin, Hyun Jung;Kim, Wan Seok;Lee, Jong Tae;Park, Chae Kyu
    • Natural Product Sciences
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    • v.20 no.3
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    • pp.160-169
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    • 2014
  • 44 compounds and 9 minerals were isolated from and detected in the New Zealand deer velvet antler Cervus elaphus var. scoticus L$\ddot{o}$nnberg. The chemical structures of (1 - 26) were identified on the basis of the spectroscopic methods and comparisons with literature, respectively. The structures were identified as cholesterol (CS, 6), 7-keto-CS (7), $7{\beta}$-hydroxy-CS (8), and $7{\alpha}$-hydroxy-CS (9), and included 12 steroid $3{\beta}$-O-(palmitic/stearic/myristic acid esters; PM/SA/MS) [CS-$3{\beta}$-O-PM (1 - 1), CS-$3{\beta}$-O-SA (1 - 2), CS-$3{\beta}$-O-MR (1 - 3), 7-keto-CS-$3{\beta}$-O-PM (2 - 1), 7-keto-CS-$3{\beta}$-O-SA (2 - 2), 7-keto-CS-$3{\beta}$-O-MR (2 - 3), $7{\beta}$-hydroxy-CS-$3{\beta}$-O-SA (3 -1), $7{\beta}$-hydroxy-CS-$3{\beta}$-O-PM (3 - 2), $7{\beta}$-hydroxy-CS-$3{\beta}$-O-MR (3 - 3), $7{\alpha}$-hydroxy-CS-$3{\beta}$-O-SA (4 - 1), $7{\alpha}$-hydroxy-CS-$3{\beta}$-O-PM (4 - 2), and $7{\alpha}$-hydroxy-CS-$3{\beta}$-O-MR (4 - 3)], dinonyl phthalate (5), 8 nucleic acids analogues [uracil (10), deoxyguanosine (11), deoxyuridine (12), uridine (13), deoxyadenosine (14), adenosine (15), inosine (16), and guanosine (17)], and the 9 free amino acids [L-phenylalanine (18), L-isoleucine (19), L-leucine (20), L-tyrosine (21), L-valine (22), L-proline (23), L-threonine (24), L-alanine (25), and L-hydroxyproline (26)]. Also, there are 8 kinds of amino acids [asparagine, serine, glutamine, glycine, histidine, arginine, methionine, and lysine], 2 sialic acids [N-acetylneuraminic acid (27), ketodeoxynonulosonic acid (28)], and 9 minerals [Na > K > Ca > Mg > Fe > Zn > B > Al > Cu] were detected from the autoaminoacid analyzer and ICP spectrometer, HPAEC-PAD/HPLC-FLD, respectively. 9 kinds of oxycholesterol-$3{\beta}$-O-fatty acid ester (2 - 1, 2 - 2, 2 - 3, 3 - 1, 3 - 2, 3 - 3, 4 - 1, 4 - 2, and 4 - 3) and 3 nucleic acids (12, 14, and 15) were isolated from the velvet antler for the first time. 6 kinds of steroids (7, 8, 9, 2 - 1, 3 - 1, and 4 - 1) were examined for their anti-proliferative effects against L1210, P388D1, K562, MEG-01, KG-1, MOLT-4, A549, HepG2, MCF-7, SK-OV-3, and SW-620 cancer cell lines. They showed anti-proliferative effects with $IC_{50}$ values of 0.06, 2.16, 2.42, > 50.0, 1.66 and $8.31{\mu}M$ against L1210, while the values were 24.05, 9.44, 5.22, 0.25. 9.48 and $49.77{\mu}M$ against P388D1, respectively. The others were inactive.

$H_2$ Receptor-Mediated Relaxation of Circular Smooth Muscle in Human Gastric Corpus: the Role of Nitric Oxide (NO)

  • Lee, Sang Eok;Kim, Dae Hoon;Kim, Young Chul;Han, Joung-Ho;Choi, Woong;Kim, Chan Hyung;Jeong, Hye Won;Park, Seon-Mee;Yun, Sei Jin;Choi, Song-Yi;Sung, Rohyun;Kim, Young Ho;Yoo, Ra Young;Park, Hee Sun;Kim, Heon;Song, Young-Jin;Xu, Wen-Xie;Yun, Hyo-Yung;Lee, Sang Jin
    • The Korean Journal of Physiology and Pharmacology
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    • v.18 no.5
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    • pp.425-430
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    • 2014
  • This study was designed to examine the effects of histamine on gastric motility and its specific receptor in the circular smooth muscle of the human gastric corpus. Histamine mainly produced tonic relaxation in a concentration-dependent and reversible manner, although histamine enhanced contractility in a minor portion of tissues tested. Histamine-induced tonic relaxation was nerve-insensitive because pretreatment with nerve blockers cocktail (NBC) did not inhibit relaxation. Additionally, $K^+$ channel blockers, such as tetraethylammonium (TEA), apamin (APA), and glibenclamide (Glib), had no effect. However, $N^G$-nitro-L-arginine methyl ester (L-NAME) and 1H-(1,2,4)oxadiazolo (4,3-A) quinoxalin-1-one (ODQ), an inhibitor of soluble guanylate cyclase (sGC), did inhibit histamine-induced tonic relaxation. In particular, histamine-induced tonic relaxation was converted to tonic contraction by pretreatment with L-NAME. Ranitidine, the $H_2$ receptor blocker, inhibited histamine-induced tonic relaxation. These findings suggest that histamine produced relaxation in circular smooth muscle of human gastric smooth muscle through $H_2$ receptor and NO/sGC pathways.

Effects of Crataegii fructus on the Contractile Response of Rabbit Corpus Cavernosum (산사(山査)가 토끼 음경해면체의 수축에 미치는 영향)

  • Lee, Han Seok;Park, Sun Young
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.27 no.5
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    • pp.602-610
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    • 2013
  • This study was aimed to evaluate the cavernosal relaxation effect of Crataegii fructus(CF) in the contracted rabbit penile corpus cavernosum by agonists.In order to study the effect of CF on the vasoconstriction of rabbit penile corpus cavernosum, isolated rabbit penile corpus cavernosum tissues were used for the experiment using organ baths containing Krebs solution.To investigate the cavernosal relaxation of CF, CF extract at $0.01{\sim}3.0mg/m{\ell}$ was added after penile corpus cavernosum were contracted by norepinephrine(NE) $1{\mu}M$. To analyze the mechanism of CF's vasorelaxation, CF extract infused into contracted penile tissues by NE after each treatment of indomethacin(IM), $N{\omega}$-nitro-L-arginine(L-NNA), methylene blue(MB), tetraethylammonium chloride(TEA).To study the effect of CF on influx of extracellular calcium chloride($Ca^{2+}$) in penile tissues, in $Ca^{2+}$-free krebs solution, $Ca^{2+}$ 1 mM infused into contracted penile tissues by NE after pretreatment of CF. Cytotoxic activity of CF on human umbilical vein endothelial cell(HUVEC) was measured by MTT assay, and nitric oxide(NO) prodution was measured by Griess reagent. CF relaxed cavernosal strip with endothelium contracted by NE, but in the strips without endothelium, CF-induced relaxation was significantly inhibited. The pretreatment of L-NNA, MB, TEA decreased significantly on the cavernosal relaxation than not-treatment of them. But the pretreatment of IM had no significant effect on the cavernosal relaxation. In $Ca^{2+}$-free krebs solution, when $Ca^{2+}$ infused into contracted penile tissues by NE, pretreatment of CF inhibit contraction induced by adding $Ca^{2+}$.NO production wasn't increased by treatment of CF on HUVEC. This findings showed that CF is effective for the relaxation of rabbit penile corpus cavernosum, and we suggest that CF relax rabbit corpus cavernosal smooth muscle through multiple action mechanisms that include increasing the release of nitric oxide from corporal sinusoidal endothelium, inhibition of $Ca^{2+}$ mobilization into cytosol from the extracellular fluid, and maybe a hyperpolarizing action.

Red Ginseng Saponin Fraction A Isolated from Korean Red Ginseng by Ultrafiltration on the Porcine Coronary Artery

  • Jung, Young-Hyun;Park, Kwang-Yeol;Jeon, Jin-Hong;Kwak, Yi-Seong;Song, Yong-Bum;Wee, Jae-Joon;Rhee, Man-Hee;Kim, Tae-Wan
    • Journal of Ginseng Research
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    • v.35 no.3
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    • pp.325-330
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    • 2011
  • Red ginseng saponin fraction-A (RGSF-A) contains a high percentage of panaxadiol saponins that were isolated from Korean red ginseng by ultrafiltration. The aim of this study was to elucidate the effects of RGSF-A on the porcine distal left anterior descending (LAD) coronary artery. The relaxant responses to RGSF-A were examined during contractions induced by 100 nM U46619 (9,11-dideoxy-9a,11a-methanoepoxy-prostaglandin F2a), a stable analogue of thromboxane A2. RGSF-A dose-dependently induced biphasic (fast- and slow-) relaxation in the distal LAD coronary artery in the presence of an intact endothelium. The fast-relaxation was quickly achieved in a minute, and then the slow-relaxation was slowly developed and sustained for more than thirty minutes after the administration of RGSF-A. The slow-relaxation had a tendency to be bigger than the fast-relaxation. Fast relaxation induced by RGSF-A was almost blocked by $N_{\omega}$-Nitro-L-arginine methyl ester (L-NAME), a nitric oxide synthase synthase inhibitor and 1H-[1,2,4]oxadiazolo[4,3-a]quinoxalin-1-one (ODQ), a guanylate cyclase inhibitor. However slow relaxation induced by RGSF-A was only partially inhibited by L-NAME and ODQ. In the endothelium-removed ring, RGSF-A evoked only slowrelaxation to a certain extent. These data suggest that RGSF-A induced both endothelium dependent fast- and slow-relaxation and endothelium independent slow-relaxation in the porcine distal LAD coronary artery. The endothelium dependent fast-relaxation is mediated by the nitric oxide (NO)-cGMP pathway, and the endothelium dependent slow-relaxation is at least partially mediated by the NO-cGMP pathway. However, the endothelium-independent slow-relaxation remains to be elucidated.

Mechanism of Hyperalgesia Following Cutaneous Inflammation by Complete Freund Adjuvant (Complete Freund Adjuvant에 의한 피부염증에서 통각과민현상의 기전)

  • Jeong, Yong;Leem, Joong-Woo;Chung, Seung-Soo;Kim, Yun-Suk;Yoon, Duck-Mi;Nam, Taick-Sang;Paik, Kwang-Se
    • The Korean Journal of Pain
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    • v.13 no.2
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    • pp.164-174
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    • 2000
  • Background: After an injury to tissue such as the skin, hyperalgesia develops. Hyperalgesia is characterized by an increase in the magnitude of pain evoked by noxious stimuli. It has been postulated that in the mechanism of hyperalgesia (especially secondary hyperalgesia) and allodynia, a sensitization of central nervous system such as spinal dorsal horn may contribute to development of hyperalgesia. However, the precise mechanism is still unclear. In the present study, we investigated the roles of N-methyl-D-aspartate (NMDA) receptor and nitric oxide (NO) system in the mechanism of hyperalgesia, and their relations with c-fos expression Methods: Inflammation was induced by injection of complete Freund adjuvant (CFA) into unilateral hindpaw of Sprague-Dawley rat. Behavioral studies measuring paw withdrawal responses by von Frey filaments and paw withdrawal latencies by radiant heat stimuli and stainings of nicotinamide adenine dinucleotide phosphate (NADPH)-diaphorase and c-fos immunoreactivity were performed. The effects of MK-801, an NMDA receptor blocker and $N^\omega$-nitro-L-arginine (L-NNA), a nitric oxide synthase (NOS) inhibitor were evaluated. Results: 1) Injection of CFA induced mechanical allodynia, mechanical hyperalgesia and thermal hyperalgesia. And it increased the number of NADPH-diaphorase positive neurons and c-fos expression neurons. 2) MK-801 inhibited mechanical hyperalgesia and thermal hyperalgesia induced by CFA and reduced the number of NADPH-diaphorase positive neurons and c-fos expression neurons. 3) L-NNA inhibited the thermal hyperalgesia and reduced the number of NADPH-diaphorase positive neurons, but did not affect the number of c-fos expression neurons. Conclusions: These results suggest that in the mechanism of mechanical hyperalgesia, NMDA receptor but not NO-system is involved and in the case of thermal hyperalgesia both NMDA receptor and NO system are involved. NO system did not affect the expression of c-fos, but c-fos expression and NOS activity were dependent on the activity of NMDA receptor.

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Ethanol Extract of Cynanchum wilfordii Produces Endothelium-Dependent Relaxation in Rat Aorta and Anti-inflammatory Activity in Human Aortic Smooth Muscle Cells

  • Choi, Deok-Ho;Lee, Yun-Jung;Kim, Eun-Joo;Li, Xiang;Kim, Hye-Yoom;Hwang, Sun-Mi;Yoon, Jung-Joo;Lee, So-Min;Min, Eun-Kyeong;Kang, Dae-Gill;Lee, Ho-Sub
    • The Journal of Korean Medicine
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    • v.31 no.6
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    • pp.47-57
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    • 2010
  • Objective: The present study investigated the effect of ethanol extract of Cynanchum wilfordii (ECW) on vascular relaxation and vascular inflammation in rat artery isolated from rats and anti-inflammatory activity in human aortic smooth muscle cells (HASMC). Methods: Vascular tone and guanosine 3',5'-cyclic monophosphate (cGMP) production were examined in rat artery isolated from Sprague Dawley rats, in the presence of ECW. HASMC were incubated with tumor necrosis factor-alpha (TNF-${\alpha}$) or Angiotensin II for 24 h. Matrix metalloproteinase (MMP)-2 and anti-oxidant activity of ECW was investigated by pretreatment with ECW in HASMC. Results: Cumulative treatment of ECW relaxed aortic smooth muscles of rats in a dose-dependent manner. ECW-induced vasorelaxation was significantly decreased by pretreatment of L-arginine methyl ester (L-NAME) or oxadiazolo-quinoxalinone (ODQ). Furthermore, ECW treatment of thoracic aorta significantly increased cGMP production. Incubation of ECW with ODQ or L-NAME markedly decreased ECW-induced cGMP production. ECW treatment dose-dependently suppressed TNF-${\alpha}$- or Angiotensin II-induced increase in matrix metalloproteinase-2 expression in HASMC. Also, ECW exhibited 2,2-diphenyl-1-picrylhydrazyl radical scavenging activity in vitro and reduced TNF-${\alpha}$-induced increase in reactive oxygen species production in a dose-dependent manner. Conclusions: Taken together, the results suggest that ECW exerts vascular relaxation via NO/cGMP signaling pathway and decreases MMP-2 expression via anti-oxidant activity.

Relaxation Effects of Rubus coreanus in Isolated Rabbit Corpus Cavernosum Smooth Muscle (복분자(覆盆子)의 토끼 음경해면체 평활근 이완효과)

  • Park, Sun Young;Lee, Pyeng Jae;Shin, Seon Mi;Kim, Ho Hyun
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.27 no.4
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    • pp.400-408
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    • 2013
  • This study aimed to investigate the relaxation effects and its underlying mechanisms of Rubus coreanus(RC) extract in contracted rabbit corpus cavernous tissues by phenylephrine(PE) $1{\mu}M$. In order to define the relaxation effects of RC, rabbit corpus cavernous tissues were prepared in $2{\times}2{\times}6mm$ sized strip. The dose-dependent relaxation responses of RC at 0.01-3.0 $mg/m{\ell}$ in contracted strips induced by PE were measured and also observed after endothelial denudation. To analyze the underlying mechanisms of RC-induced relaxation, indomethacin(IM), tetraethylammonium chloride(TEA), $N{\omega}$-nitro-L-arginine (L-NNA), methylene blue(MB) were treated before RC extract infused into precontracted strips induced by PE. To study the effect of RC extract on influx of extracellular $Ca^{2+}$ in corpus cavernous strips, calcium chloride(Ca) 1 mM infused into precontracted strips induced by PE after pretreatment of RC extract in $Ca^{2+}$-free krebs-ringer solution. To investigate cytotoxic activity and nitric oxide(NO) concentration of RC extract on human umbilical vein endothelial cell(HUVEC), cell viability on HUVEC was measured by MTT assay, and NO concentration was measured by Griess reagent system. The cavernous strips were significantly relaxed by RC extract at 1.0 $mg/m{\ell}$, 3.0 $mg/m{\ell}$ and the relaxation responses to RC were inhibited significantly by endothelial disruption. The pretreatment of IM, TEA didn't affect RC extract-induced endothelium-dependent relaxation, but the pretreatment of L-NNA, MB reduced RC extract-induced endothelium-dependent relaxation. When $Ca^{2+}$ was supplied the cavernous strips which were precontracted by PE in a $Ca^{2+}$-free krebs-ringer solution, contraction of strips was increased, but pretreatment of RC inhibited contractile response to $Ca^{2+}$. When RC extract was applicated on HUVEC, NO concentration was increased. Our findings show that RC extract exerts a relaxing effect on corpus cavernosum in part by suppressing influx of extracellular $Ca^{2+}$ through activating the NO-cGMP system.