• Journal of Ginseng Research
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• 제16권3호
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• pp.175-182
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• 1992

Intravenous administration of saponin from the root of Panax ginseng (red ginseng) lowered the blood pressure in a dose-dependent manner (10~100 mg/kg B.W) in anesthetized rats. Therefore, experiments were designed to study whether this lowering of blood pressure is associated with the release of endothelium-derived relaxing factor. Rings of thoracic aorta with and without endothelium were suspended for the measurement of isometric tension in organ chamber. All experiments were performed in the presence of indomethacin (10-5 M). Ginseng saponin (10-5~3$\times$10-4 g/ml) relaxed contractions induced by phenylephrine (10-5 M) in the aorta with endothelium but not in that without endothelium. Treatment of aortic rings with NG_monomethyl-L-arginine (L-NMMA 10-4 M for 30 min), a competive inhibitor of nitric oxide synthase and methylene blue (M.B., 3$\times$10-7 M for 30 min), an inhibitor of soluble guanylate cyclase, diminished the relaxation induced by ginseng saponin. In thoracic aortic rings from rats treated with ginseng saponin for 2 weeks intraperitoneally, the relaxation to acetylcholine was increased compared with non-ginseng treated rings. These data suggest that red ginseng saponin evokes hypotension and that vascular relaxations induced by red ginseng saponin are inediatpd by release of endothelium-derived relaxing factor.

• 대한한의학회지
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• 제18권2호
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• pp.15-32
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• 1997

This study was undertaken to define the mechanism of Siegesbeckiae Herba-induced relaxation in rabbit common carotid artery contracted by agonists. In order to investigate the effect of Siegesbeckiae Herba on contracted rabbit carotid arterial strips, transverse strips with intact or damaged endothelium were used for the experiment using organ bath. To analyze the mechanism of Siegesbeckiae Herba-induced relaxation, Siegesbeckiae Herba extract infused into contracted arterial strips induced by agonists after treatment of lanthanum chloride, indomethacin, atropine, $N\omega-nitro-{_L}-arginine$, cobalt chloride or methylene blue. The relaxation effect of Siegesbeckiae Herba was dependent on the presence of endothelium, showing that Siegesbeckiae Herba-induced relaxation was not observed in the strips without endothelium. The endothelium-dependent relaxation induced by Siegesbeckiae Herba was suppressed by the pretreatment of lanthanum chloride, $N\omega-nitro-{_L}-arginine$, cobalt chloride or methylene blue, but it was not observed in the strips pretreated with indomethacin or atropine. These results demonstrated that Siegesbeckiae Herba may inhibit agonist-induced contraction through an increase in the cyclic GMP by the production of nitric oxide in the vascular endothelial cells.

• 고려인삼학회:학술대회논문집
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• 고려인삼학회 1993년도 학술대회지
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• pp.40-48
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• 1993

Intravenous administration of saponin extracted from the root of Panax ginseng lowered the blood pressure dose-dependently (10-200 mg/kg, B.W) in anesthetized rats. Therefore, experiments were designed to study the hypothesis that the lowering of blood pressure is associated with the release of endothelium-derived relaxing factor and the accumulation of guanosine 3, 5-cyclic monophosphate (cGMP). Rings of thoracic aorta with and without endothelium were suspended for the measurement of isometric tension in organ chamber and the tissue content of cGMP was measured by radioimmunoassay. All experiments were performed in the presence of $indomethacin(10^{-5}M).$ Ginseng saponin $(10^{-5}-3{\times}10^{-6}g/ml)$ relaxed contractions induced by phenylephrine $10^{-6}M)$ in the aorta with endothelium but not in that without endothelium. Treatment of aortic rings with $N^G$ monomethyl-L-arginine (L-NMMA, $10^{-4}M$ for 30 min), a competitive inhibitor of nitric oxide synthase, and methylene blue $(MB,\;3{\times}10^{-7}M$ for 30 min). an inhibitor of soluble guanylate cyclase, diminished the relaxation induced by Ginseng saponin. Ginseng saponin $10^{-4}g/ml$ for 2 min) increased the accumulation of cGMP in rings with endothelium. L-NMMA and MB inhibited the accumulation of cGMP induced by Ginseng saponin. These data suggest that vascular relaxations induced by Ginseng saponin are mediated by release of endothelium-derived relaxing factor and the accumulation of cGMP. The effect of Ginseng saponin on endothelial function in hypercholesterolemic rabbits was examined. In hypercholesterolemic rabbits fed with $2\%$ cholesterol for 8 weeks, relaxation of aortic rings to acetylcholine was impaired. The impaired relaxations of aortic rings in hypercholesterolemic rabbits were improved by dietary supplementation of Ginseng saponin, probably because of an improved release of endothelium - derived relaxing factor.

• 대한약리학회지
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• 제27권2호
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• pp.125-133
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• 1991

내피세포가 제거된 토끼의 적출 장간막 동맥에서 토끼의 대동맥 내피세포로 부터 A23187과 acetylcholine은 NO와 유사한 혈관 이완성 물질 (EDRF)을 유리한다. 이에 첨가하여 A23187은 acetylcholine과는 달리 superoxide anion에 의하여 파괴되지 않는 EDRF도 유리시킴을 확인하고 이의 특성에 대하여 연구하였다. 정상적인 생리영양액에서는 A23187과 acetylcholine의 용량-반응 곡선은 내피세포에 의존하지 않는 sodium nitroprusside의 곡선과 유사하였다. 이들은 methylene blue에 의하여 억제되었다. Hypoxanthine (HX)과 xanthine oxidase (XO)를 bath내로 투여시 phenylephrine에 의한 수축이 일과성으로 증가한 후 지속적으로 이완되었다. HX-XO 반응중에는 A23187은 장간막 동맥을 즉각적으로 이완시켰으나 acetylcholine의 이완작용은 소실되었다. A23187에 의하여 야기되는 장간막동맥의 이완은 50 mM $K^+-PSS$로 수축을 야기시켰을 때에는 나타나지 아니하였다. Superoxide dismutase를 전처치하였을 때는 HX-XO 반응중에도 acetylcholine 뿐만아니라 A23187에 의한 장간막 동맥의 수축은 이완되었다. 한편, acetylcholine에 의하여 야기되는 장간막 동맥의 이완은 A23187에 의하여 야기되는 이완에 비하여 phorbol 12-myristate 13-acetate (PMA)에 훨씬 더 민감하게 억제되었다. 내피세포의 기능과는 무관한 sodium nitroprusside에 의하여 야기되는 이완은 PMA에 의하여 영향을 받지 아니하였다. 이상의 결과로 미루어 볼때, A23187과 acetylcholine은 methylene blue에 의하여 억제되는 내피세포 의존성 이완을 야기시키고, 첨가하여 A23187은 어떤 병적 환경 아래서는 superoxide anion과 PMA에 저항성을 지닌 혈관이완성 물질을 유리하는 것으로 사료된다. 앞으로 A23187에 의하여 유리되는 혈관 이완성 물질이 superoxide anion에 의존하여 생성된 것인지에 대하여는 추후의 연구과제이다.

• Journal of Chest Surgery
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• 제24권3호
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• pp.229-244
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• 1991

A bioassay technique and organ bath study were performed to analyze the effects of extracellular $Ca^{2+}$ and $Ca^{2+}$-antagonists on endothelium-derived relaxing factor[s][EDRF] released from the endothelial cells of rabbit aorta. Transverse strips with intact endothelium or damaged endothelium were used for the mechanical contraction experiment using organ bath. Long segment including thoracic and abdominal aorta with endothelium [EDRF donor aorta] was perfused with Tyrode solution which was aerated with 95% $O_2-5%$ $CO_2$ mixed gas and kept at 35oC. The perfusate was bioassayed with a transverse strip of thoracic aorta with damaged endothelium. The test strip was contracted with nor-epinephrine and acetylcholine was used to stimulate the release of EDRF from endothelial cells. The results obtained were as follows; 1] The endothelium-dependent relaxation[EDR] induced by acetylcholine was biphasic; an initial rapid relaxation followed by a slow relaxation. 2] EDR induced by acetylcholine was reduced gradually with the decrease in the concentration of extracellular $Ca^{2+}$. The effect of extracellular $Ca^{2+}$ on EDR was more prominent in the late slow relaxation phase. 3] EDR to acetylcholine was not altered by acute exposure to organic $Ca^{2+}$-antagonists. Pretreatment with verapamil to the EDRF donor aortic segment did not alter the magnitude of EDR. 4] Among the inorganic $Ca^{2+}$-antagonists $Mn^{2+}$ and $Cd^{2+}$ did not inhibit EDR, whereas $Co^{2+}$ and $La^{3+}$ inhibited EDR. 5] The inhibitory response of $Co^{2+}$ to EDR developed when infused directly on the test strip. That of $La^{3+}$, however, was evoked when added to solution perfusing the donor aortic segment. The above results suggest that $Ca^{2+}$-antagonists do not affect EDR and the inhibitory effect of $Ca^{2+}$ results from influencing the action of EDRF on vascular smooth muscle, whereas that of $La^{3+}$ results from its action on the release of EDRF from endothelial cells.

• 대한수의학회지
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• 제44권3호
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• pp.357-366
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• 2004

We studied the effect of propofol (PPF) on the endothelium-dependent vascular responses in isolated rat thoracic aorta. In aortic rings with endothelium, PPF inhibited the phenylephrine (PE)-induced contraction in a concentration-dependent manner. In PE-precontracted preparations, PPF attenuated the endothelium-dependent relaxation by acetylcholine but not by A23187. And PPF did not attenuate the endothelium-independent relaxation by sodium nitroprusside (SNP). The relaxation induced by acetylcholine in PE-precontracted aortic rings was significantly augmented by zaprinast, a cGMP-specific phosphodiesterase inhibitor, and this augmentation was inhibited by PPF. Although SNP-induced relaxation was significantly augmented by zaprinast, this augmentation was not inhibited by PPF. In preparations preconstricted with PE, the PPF-induced relaxation was inhibited by atropine. In addition, PPF attenuated the vasorelaxation by phosphodiesterase inhibitors (IBMX, Ro20-1724 or zaprinast except milrinone). In vivo, the infusion of acetylcholine and SNP showed decreased arterial blood pressure in rats. The pre-injection of PPF inhibited the acetylcholine-induced blood pressure lowering, but not the SNP-induced blood pressure lowering. These results suggest that PPF can attenuate in part the acetylcholine-induced vasorelaxation and blood pressure lowering through the inhibition of the acetylcholine receptor-mediated endothelium-derived relaxing factor by acting on endothelium. It is considered that the inhibitory effect of PPF on the vasorelaxation is due to the decreased level of cGMP which can be attributed to the inhibition of the muscarinic receptor and/or receptor-G-protein interaction.

• 동의생리병리학회지
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• 제27권6호
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• pp.809-817
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• 2013

The aim of this study was to evaluate the mechanism of vasodilation of Lespedezea cuneata(LC) in rabbit common carotid artery and cavernosal smooth muscle. LC relaxed arterial strips precontracted with norepinephrine and cavernosal strips precontracted with phenylephrine. The arterial relaxation effects of LC was endothelium-dependent. $N{\omega}$-nitro-L-arginine(L-NNA), NOS inhibitor, methylene blue(MB), cGMP inhibitor, indomethacin(IM), cyclo-oxygenase inhibitor and tetraethylammonium chloride(TEA), KCa-channel blocker attenuate the relaxation responses of LC in arterial strips. In $Ca^{2+}$-free krebs-ringer solution, pretreatment of LC extract significantly reduced the contraction induced by addition $Ca^{2+}$. L-NNA reduced LC extract-induced relaxation in cavernosal strips, but IM, TEA and MB didn't affect LC extract-induced relaxation. When LC extract was applicated on human umbilical vein endothelial cell, the nitric oxide concentration was increased. We conclude that in rabbit common carotid artery, LC may suppress influx of extra-cellular $Ca^{2+}$ through the release of endothelium derived relaxing factor including nitric oxide, prostacyclin, endothelium derived hyperpolarizing factor. And LC exerts a relaxing effect on corpus cavernosum through activating the NO.

• The Korean Journal of Physiology
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• 제24권1호
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• pp.91-102
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• 1990

토끼 흉부 대동맥을 이용하여 내피세포 의존성 혈관이완에 대한 세포외 $Ca^{2+}$과 여러가지 $Ca^{2+}$ 길항제의 효과를 분석하여 EDRF의 작용기전을 밝혀 보고자 하였다. 대동맥 횡단 절편의 등장성 수축은 $10^{-7}\;M$ 노에피네프린으로 유발시켰으며, $10^{-6}\;M$ 사세틸콜린으로 내피세포 의존성 혈관이완을 일으켰다. 내피세포는 작은 솜뭉치로 부드럽게 문질러서 제거하였으며, hemolysate를 사용하여 EDRF에 대한 헤모글로빈의 효과를 관찰하였다. 결과를 종합하면 다음과 같다. 1) 아세틸콜린에 의한 내피세포 의존성 혈관이완은 두 시기, 즉 초기급속이완기와 후기완만이완기로 나타났다. 2) 세포외 $Ca^{2+}$을 낮추면, 아세틸콜린에 의한 내피세포 의존성 혈관이완이 감소하였으며, 특히 후기완만이완기가 감소하였다. 3) Verapamil, nifedipine, $Mn^{2+}$ 및 $Cd^{2+}$은 내피세포 의존성 혈관이완에 영향이 없었던 반면 $La^{3+}$와 $Co^{2+}$는 억제시켰다. 4) 헤모글로빈을 투여하면 내피세포가 없는 절편에서는 기초긴장도의 변화가 없었으나 내피세포가 있는 절편에서는 기초긴장도가 증가하였고 아세틸콜린에 의한 내피세포 의존성 혈관이완도 완전히 억제되었다. 이상의 결과로부터 세포외 $Ca^{2+}$은 주로 후기완만이완기에 작용하며 이때 사용되는 $Ca^{2+}$ 유입 통로는 $Ca^{2+}$ 길항제로 억제되지 않는 것으로 결론지을 수 있다.

• The Korean Journal of Physiology
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• 제28권2호
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• pp.181-190
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• 1994

Endothelium-derived relaxing factor (EDRF) activates guanylate cyclase which mediates the formation of cGMP from GTP in vascular smooth muscle. It is well known that endothelium-dependent relaxation is impaired in spontaneously hypertensive rats (SHR). However, it is still unknown whether the impaired endothelium-dependent relaxation in SHR results from the reduced release of EDRF or from the decrease of vascular response to EDRF. We investigated the effects of cGMP on the contractility and Ca movement in the aorta of SHR and Wistar-Kyoto rats (WKY). The amplitude of the endothelium-dependent relaxation to actylcholine (ACh) was significantly less in SHR than in WKY. L-arginine $(10^{-3}M)$ did not increase endothelium-dependent relaxation in both strains. Sodium nitroprusside (SNP), an activator of guanylate cyclase, relaxed the 40 mM $K^+-induced$ contraction in a dose-dependent manner $(10^{-10}{\sim}10^{-6}\;M)$ in the endothelium-rubbed aortic strips of both strains. However, there was no significant difference in these relaxations between WKY and SHR. 8-bromo-cyclic guanosine monophosphate (8-Br-cGMP), a cell membrane-permeable derivative of cGMP relaxed the 40 mM $K^+-induced$ contraction in a dose-dependent manner $(10^{-6}{\sim}10^{-4}\;M)$ in the endothelium-rubbed aortic strips of both strains. Also norepinephrine $(10^{-6}\;M)-induced$ contractions in normal and Ca-free Tyrode's solution were suppressed by the pretreatment with 8-Br-cGMP $(10^{-4}\;M)$ in either strain. However, the amplitudes of suppression induced by 8-Br-cGMP were greater in SHR than that in WKY. Basal $^{45}Ca$ uptake and 40mM $K^+-stimulated\;^{45}Ca$ uptake were not suppressed by pretreatment with 8-Br-cGMP $(10^{-4}\;M)$ in single aortic smooth muscle cells of both SHR and WKY. From the above results, it is suggested that cGMP decreases Ca sensitivity in vascular smooth muscle cells and that the impaired endothelium-dependent relaxation in the aortic strips of SHR is not the result of a reduced vascular response to EDRF.

• The Korean Journal of Physiology
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• 제27권1호
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• pp.27-35
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• 1993

Effects of the endothelium on the contractile responses to norepinephrine (NE) were investigated in isolated helical strips of the proximal and distal coronaries artery of pigs. The helical strips were immersed in Tris-buffered Tyrode's solution equilibrated with 100% $O_2$ at $35^{\circ}C$ and its isometric tension was measured. NE relaxed the strips precontracted with acetylcholine from both the proximal and distal coronary artery. NE-induced relaxation, which might be induced mainly by $\beta$-adrenoceptor function was dominant in the distal coronary arteries. NE-induced relaxation was converted to a contraction after $\beta$-adrenoceptor blockade with propranolol $(3{\times}10^{-6}M)$. ${\alpha}$-adrenoceptor-mediated contraction by NE was greater in the proximal coronary artery than the distal coronary artery. Quantitatively, ${\alpha}_1$-adrenoceptor mediated contraction by NE was greater than ${\alpha}_2$-adrenoceptor mediated contraction by NE in both arteries. NE-induced relaxation was decreased by rubbing of endothelium in both arteries. ${\alpha}_1-and\;{\alpha}_2$-adrenoceptor mediated contraction by NE were potentiated by rubbing of endothelium in both arteries. Pretreatment with methylene blue, an inhibitor of soluble guanylate cyclase, increased ${\alpha}_1-\;and\;{\alpha}_2$-adrenoceptor mediated contraction by NE in both arteries with endothelium. From the above results, we suggest that the effect of activation of $\alpha$-adrenoceptors by NE may be modulated by endothelium in the proximal and distal coronary arteries of pigs. This effect may be mediated via endothelium derived relaxing factor.