Release of a Stable Endothelium-derived Relaxing Factor by A23187 from the Rabbit Aortic Endothelium

김치대,임병용,홍승철,홍기환,Kim, Chi-Dae,Rhim, Byung-Yong,Hong, Sung-Chul,Hong, Ki-Whan The Korean Society of Pharmacology 1991 대한약리학잡지 Vol.27 No.2

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. In the isolated rabbit mesenteric artery denuded of endothelium, we characterized the identity of the A23187-induced endothelium-dependent relaxing factor (EDRF) released from the endothelium of rabbit aorta, which is distinct from that of acetylcholine-induced relaxing factor. In the normal physiological salt solution (PSS), the dose-response curves to A23187 and acetylcholine were overlapped together. Their effects were also inhibited by methylene blue. Upon application of hypoxanthine and xanthine oxidase into the bath, the phenylephrine-induced precontraction was transiently increased followed by the sustained relaxation. During the burst of hypoxanthine-xanthine oxidase reaction, the $Ca^{++}$ ionophore, A23187 but not acetylcholine was able to cause an immediate relaxation. However, A23187-induced relaxation was not manifested when precontracted by 50 mM $K^+-PSS$. Nevertheless, in the presence of superoxide dismutase, A23187 could produce an immediate relaxation without accompanying the transient contraction as acetylcholine did during the hypoxanthine-xanthine oxidase reaction. On the other hand, acetylcholine-induced relaxation was more sensitively inhibited by phorbol 12-myristate 13-acetate (PMA) than A23187-induced relaxation. Endothelium-independent relaxation to sodium nitroprusside was not affected by PMA. Based on these results it is suggested that both A23187 and acetylcholine cause the methylene blue-inhibitable endothelium-dependent relaxation, and in addition, A23187 may release a stable EDRF which is resistant to superoxide anion and PMA.

토끼 대동맥 내피에서 A23187에 의하여 유리되는 혈관이완물질의 특성에 관한 연구

김치대(Chi-Dae Kim),임병용(Byung-Yong Rhim),홍승철(Sung-Chul Hong),홍기환(Ki-Whan Hong) 대한약리학회 1991 대한약리학잡지 Vol.27 No.2

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⁺-PSS로 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. In the isolated rabbit mesenteric artery denuded of endothelium, we characterized the identity of the A23187-induced endothelium-dependent relaxing factor (EDRF) released from the endothelium of rabbit aorta, which is distinct from that of acetylcholine-induced relaxing factor. In the normal physiological salt solution (PSS), the dose-response curves to A23187 and acetylcholine were overlapped together. Their effects were also inhibited by methylene blue. Upon application of hypoxanthine and xanthine oxidase into the bath, the phenylephrine-induced precontraction was transiently increased followed by the sustained relaxation. During the burst of hypoxanthine-xanthine oxidase reaction, the Ca⁺⁺ ionophore, A23187 but not acetylcholine was able to cause an immediate relaxation. However, A23187-induced relaxation was not manifested when precontracted by 50 mM K⁺-PSS. Nevertheless, in the presence of superoxide dismutase, A23187 could produce an immediate relaxation without accompanying the transient contraction as acetylcholine did during the hypoxanthine-xanthine oxidase reaction. On the other hand, acetylcholine-induced relaxation was more sensitively inhibited by phorbol 12-myristate 13-acetate (PMA) than A23187-induced relaxation. Endothelium-independent relaxation to sodium nitroprusside was not affected by PMA. Based on these results it is suggested that both A23187 and acetylcholine cause the methylene blue-inhibitable endothelium-dependent relaxation, and in addition, A23187 may release a stable EDRF which is resistant to superoxide anion and PMA.

Effect of Acetylcholine on Calcium-Induced Contraction in Isolated Mouse Duodenum

Kim, Mi-Joo,Kim, In-Kyeom,Sohn, Uy-Dong,Kim, Choong-Young 慶北大學校 醫科大學 1990 慶北醫大誌 Vol.31 No.4

마우스의 적출 십이지장을 사용하여 calcium을 제거한 영양액에서 CaCl_2를 첨가할 때 나타나는 수축반응에 대한 acetylcholine의 작용을 관찰하고, atropine과 KCl 존재 여부에 따른 변화를 비교하였던 바 다음과 같은 결과를 얻었다. CaCl_2 첨가로 인해 나타나는 수축반응에 대해 acetylcholine은 용량 의존적으로 억제작용이 강하게 나타났으며, 특히 phasic contraction보다 tonic contraction에 대한 억제작용이 뚜렷했으며 atropine 존재하에서는 acetylcholine의 억제작용이 나타나지 않았다. 저농도의 KCl용액(20 mM)은 CaCl_2 첨가에 의한 수축반응 뿐 아니라 acetylcholine의 억제작용에 전혀 영향을 미치지 않았지만 고농도의 KCl용액(80 mM)은 CaCl_2첨가에 의한 수축반응을 억제시켰으며 acetylcholine과는 상협적으로 작용하였다. 이상의 결과로 미루어 calcium을 제거한 영양액에서 CaCl_2 첨가로 나타나는 수축반응에 대하여 acetylcholine은 억제작용을 나타내며 이는 muscarinic 수용체와 관련이 있는 것 같다. In an attempt to evaluate the effect of acetylcholine on the calcium-induced contraction in the isolated mouse duodenum, the contractile response which was produced by addition of 1.8 mM calcium in the calcium-depleted medium was compared with that in the presence of acetylcholine, atropine, low or high potassium solution, or two of them. Acetylcholine inhibits the contractile response produced by addition of 1.8 mM calcium in the calcium-depleted medium and its inhibitory action was dose-dependently increased and completely reversed by the presence of atropine. The calcium-induced contractile response was also inhibited in calcium-depleted, high potassium(80 mM)-depolarizing mdium and this inhibition was marked in the presence of acetylcholine but calcium-depleted, low potassium(20 mM) medium did not affect the contractile response. These results suggest that acetylcholine inhibit calcium-induced contractile response, paticularly tonic component, via muscarinic recetors.

Acetylcholine및 Oxytocin에 의하여 야기되는 렛드 자궁수축에 미치는 Verapamil 및 Tetracaine의 영향

이만기(Maan Gee Lee),김중영(Choong Young Kim) 대한약리학회 1987 대한약리학잡지 Vol.23 No.2

Acetylcholine및 oxytocin에 의하여 야기되는 흰쥐의 적출자궁수축을 Ca²⁺ 길항제의 일종인 verapamil및 tetracaine 존재하에서 acetylcholine및 oxytocin에 의한 자궁수축곡선을 4개의 요소 (trough tension, T: peak tension, P; contraction frequency, F: duration, D)로 나누어 비교분석하여 다음과 같은 결과를 얻었다. Verapamil (0.25μM)은 자발수축을 억제시켰으나 tetracaine(42μM)은 자발수축을 억제시키지 못하였다. 이들 길항제의 존재하에서 acetylcholine및 oxytocin에 의하여 야기되는 자궁수축의 각 구성요소에 변화를 관찰하였다. 즉 acetylcholine에 의한 수축에서 verapamil은 P와 D를 감소시켰고 tetracaine은 F를 감소시키고 D를 증가시켰다. oxytocin에 의한 수축에서 verapamil은 P와 D를 감소시켰으나 tetracaine은 oxytocin농도에 따라 차이가 있었는데, 저농도의 oxytocin에 의한 수축에서는 F를 감소시키고 D를 증가시켰으나 고농도의 oxytocin에 의한 수축에서 는 F와 D에는 영향을 주지 않고 P만 감소시켰다. 이상의 결과로 미루어 acetylcholine및 oxytocin에 의하여 야기되는 수축곡선은 시각적으로 큰 차이가 없었으나 작용기전이 다른 Ca²⁺ 길항제에 의하여 acetylcholine및 oxytocin의 수축의 구성요소에 다르게 영향을 미칠 수 있었다는 것은 수축곡선의 구성요소의 변화를 면밀히 검토하면 자궁수축제의 수축작용기전이 다름을 예측할 수 있을 뿐만 아니라 수축억제제에 의한 억제 기전의 차이점도 예측할 수 있을 것으로 생각되어 진다. The effects of verapamil and tetracaine on acetylcholine-and oxytocin-induced contraction of uterus from estrogen-treated rat were examined. Isometric tensions were recorded on the Physiograph and stored in TriGem 20XT computer as digitized data for off-line analysis of the components, which described the contraction patterns: trought tension (T), peak tension (P), contraction frequency (F), and duration (D). In the acetylcholine-induced contraction, verapamil (0.25μM) significantly decreased P and D. In contrast, tetracaine (42μM) decreased F, but increased D. In the low oxytocin-induced contraction, verapamil (0.25μM) decreased P and D, and tetracaine (42μM) decreased F but increased D. In the high oxytocin-induced contraction, verapamil decreased P and D, but tetracaine decreased P without affecting on other components. These results suggest that the analysis of effects of a certain inhibitor on the components of contraction allow to postulate its specific inhibitory mechanism of the smooth muscle contraction.

Effect of Dance Sports on Neurotransmitter in Transporter Imaging and Brain Blood in Senior

Chui Ho Shin,Han Yong Lee 한국발육발달학회 2010 한국발육발달학회지 Vol.18 No.4

Effect of dance sports on Acetylcholine and brain blood in senior senile. It is known that as for the old people, the Acetylcholine change caused by exercises has an effect on the blood vessel of a brain and that it also plays an important role in the prevention and medical treatment of senile dementia. The "MMSE-K" test report was used in this study. So the testosterone, Acetylcholine was analyzed in the blood for 2 weeks, divided into 4 terms. By selecting one male subject, the SPECT was used to take pictures of their brain. The results are as followings. Dance sport didn``t cause detectable changes in the Acetylcholine of man. When the integrating the results, is was possible to notice the Acetylcholine in men didn``t show significant changes until the 4th term. We consi! der that the short term exercise didn``t affect the changes of Acetylcholine. However, after adapting to the long term exercise, it seemed to be partial effective. So the increase of maintenance of Acetylcholine, which is the cause of vessel expansion, is thought to increase the brain blood and effective in preventing brain vein diseases. When the integrating the results, it was possible to notice the Acetylcholine in men didn``t show significant changes. So the increase of maintance of Acetylcholine, which is the cause of vessel expansion, is thought to increase the brain blood and effect in preventing brain vein disease.

토끼 경동맥에서 Acetylcholine의 작용에 대한 Phorbol Ester의 조절 작용

노용래,이상호,이영우 대한신경외과학회 1994 Journal of Korean neurosurgical society Vol.23 No.12

Authors studied the regulatory mechanism of protein kinase C on the action of acetylcholine in rabbit carotid artery. The arterial rings were myobphied isometrically in an isolated organ bath. In this study, acetylcholine relaxed phenylephrine-induced contraction of rabbit carotid artery in the presence of endothelium. In the pretreatment of methylene blue or nitro-L-arginine, the action of acetylchioline was reduced. Pretreatment of phorbol 12-myristate IZacetate(PMA) attenuated the action of acetylcholine, but PMA did not attenuated it in the presence of staurosporine, suggesting that protein kinase C suppressed the action of acetylcholine. The potency of phorbol ester on the action of acetylcholine was PMA>phohol 1213dibutyrate(PDBu) >phorbol 12,13-diacetate(PDA), but the direct effect of phorbol on the contraction of arterial rings was PDBu>PMA>PDA. This implied that protein kinase C involved in the contraction of smooth muscle and the attenuation of the action of acetylcholine were different PMA did not affect on A23187- and sodium nitroprusside-induced vasorelaxation. Acetylcholine increased tissue cGMP contents, which was reduced by PMA, These results suggest that in rabbit carotid artery protein kinase C reduce acetylcholine-stimuated endothelium derived relaxing factor(EDRF) release by affecting membrane receptor, and do not affect on the function of EDRF and cGMP production in the smooth muscle.

Effect of Dance Sports on Neurotransmitter in Transporter Imaging and Brain Blood in Senior

신철호,이한용 한국발육발달학회 2010 한국발육발달학회지 Vol.18 No.4

Effect of dance sports on Acetylcholine and brain blood in senior senile. It is known that as for the old people,the Acetylcholine change caused by exercises has an effect on the blood vessel of a brain and that it also plays an important role in the prevention and medical treatment of senile dementia. The “MMSE-K” test report was used in this study. So the testosterone, Acetylcholine was analyzed in the blood for 2 weeks, divided into 4 terms. By selecting one male subject, the SPECT was used to take pictures of their brain. The results are as followings. Dance sport didn't cause detectable changes in the Acetylcholine of man. When the integrating the results, is was possible to notice the Acetylcholine in men didn't show significant changes until the 4th term. We consi! der that the short term exercise didn't affect the changes of Acetylcholine. However, after adapting to the long term exercise, it seemed to be partial effective. So the increase of maintenance of Acetylcholine, which is the cause of vessel expansion, is thought to increase the brain blood and effective in preventing brain vein diseases. When the integrating the results, it was possible to notice the Acetylcholine in men didn't show significant changes. So the increase of maintance of Acetylcholine, which is the cause of vessel expansion, is thought to increase the brain blood and effect in preventing brain vein disease.

Effects of Verapamil and Tetracaine on Acetylcholine-and Oxytocin-induced Uterine Contraction Pattern

이만기,김중영,Lee, Maan-Gee,Kim, Choong-Young The Korean Society of Pharmacology 1987 대한약리학잡지 Vol.23 No.2

The effects of verapamil and tetracaine on acetylcholine-and oxytocin-induced contraction of uterus from estrogen-treated rat were examined. Isometric tensions were recorded on the Physiograph and stored in TriGem 20XT computer as digitized data for off-line analysis of the components, which described the contraction patterns: trought tension (T), peak tension (P), contraction frequency (F), and duration (D). In the acetylcholine-induced contraction, verapamil $(0.25\;{\mu}M)$ significantly decreased P and D. In contrast, tetracaine $(42{\mu}M)$ decreased F, but increased D. In the low oxytocin-induced contraction, verapamil $(0.25\;{\mu}M)$ decreased P and D, and tetracaine $(42{\mu}M)$ decreased F but increased D. In the high oxytocin-induced contraction, verapamil decreased P and D, but tetracaine decreased P without affecting on other components. These results suggest that the analysis of effects of a certain inhibitor on the components of contraction allow to postulate its specific inhibitory mechanism of the smooth muscle contraction. Acetylcholine및 oxytocin에 의하여 야기되는 흰쥐의 적출자궁수축을 $Ca^{2+}$ 길항제의 일종인 verapamil및 tetracaine 존재하에서 acetylcholine및 oxytocin에 의한 자궁수축곡선을 4개의 요소 (trough tension, T: peak tension, P; contraction frequency, F: duration, D)로 나누어 비교분석하여 다음과 같은 결과를 얻었다. Verapamil $(0.25\;{\mu}M)$은 자발수축을 억제시켰으나 tetracaine$(42\;{\mu}M)$은 자발수축을 억제시키지 못하였다. 이들 길항제의 존재하에서 acetylcholine및 oxytocin에 의하여 야기되는 자궁수축의 각 구성요소에 변화를 관찰하였다. 즉 acetylcholine에 의한 수축에서 verapamil은 P와 D를 감소시켰고 tetracaine은 F를 감소시키고 D를 증가시켰다. oxytocin에 의한 수축에서 verapamil은 P와 D를 감소시켰으나 tetracaine은 oxytocin농도에 따라 차이가 있었는데, 저농도의 oxytocin에 의한 수축에서는 F를 감소시키고 D를 증가시켰으나 고농도의 oxytocin에 의한 수축에서 는 F와 D에는 영향을 주지 않고 P만 감소시켰다. 이상의 결과로 미루어 acetylcholine및 oxytocin에 의하여 야기되는 수축곡선은 시각적으로 큰 차이가 없었으나 작용기전이 다른 $Ca^{2+}$ 길항제에 의하여 acetylcholine및 oxytocin의 수축의 구성요소에 다르게 영향을 미칠 수 있었다는 것은 수축곡선의 구성요소의 변화를 면밀히 검토하면 자궁수축제의 수축작용기전이 다름을 예측할 수 있을 뿐만 아니라 수축억제제에 의한 억제 기전의 차이점도 예측할 수 있을 것으로 생각되어 진다.

Liberation of Serotonin Is Not Unaffected by Acetylcholine in Rat Hippocampus

김재헌,안영수,송윤섭 대한배뇨장애요실금학회 2021 International Neurourology Journal Vol.25 No.S2

Purpose: Raised cerebral titers of acetylcholine have notable links with storage symptomatology related to lower urinary tract symptoms. The hippocampus contributes to the normal control of continence in the majority of instances (circuit 3). Owing to synaptic connections with other nerve cells, acetylcholine affects the micturition pathway via the liberation of additional cerebral neurotransmitters. Despite the fact that cerebral serotonin is a key inhibitor of reflex bladder muscle contractions, the influence of acetylcholine on its liberation is poorly delineated. The current research was conducted in order to explore the role of acetylcholine in serotonin liberation from sections of rat hippocampus in order to improve the comprehension of the relationship between cholinergic and serotonergic neurons. Methods: Hippocampal sections from 6 mature male Sprague-Dawley rats were equilibrated over a 30-minute period in standard incubation medium so as to facilitate [3H]5-hydroxytryptamine (5-HT) uptake. The cerebral neurotransmitter, acetylcholine, was applied to the sections. Aliquots of drained medium solution were utilized in order to quantify the radioactivity associated with [3H]5-HT liberation; any alterations in this parameter were noted. Results: When judged against the controls, [3H]5-HT liberation from the hippocampal sections remained unaltered following the administration of acetylcholine, implying that this agent has no inhibitory action on this process. Conclusions: Serotonin liberation from murine hippocampal sections is unaffected by acetylcholine. It is postulated that the bladder micturition reflex responds to acetylcholine through its immediate cholinergic activity rather than by its influence on serotonin release. These pathways are a promising target for the design of de novo therapeutic agents.

Participation of Rho-associated kinase in electrical stimulated and acetylcholine-induced contraction of feline esophageal smooth muscle

Park, S.Y.,Song, H.J.,Sohn, U.D. North-Holland ; Elsevier Science Ltd 2009 european journal of pharmacology Vol.607 No.1

The RhoA/Rho-associated kinase (ROCK) signaling pathway has been known to play a critical role in Ca<SUP>2+</SUP>-sensitization of smooth muscle contraction. In this study, we investigated the role of ROCK in feline esophageal body smooth muscle contraction induced by electrical field stimulation and exogenous acetylcholine in vitro. Y-27632 [(+)-(R)-trans-4-(1-aminoethyl)-(4-pyridyl) cyclohexanecarboxamide dihydrochloride], ROCK inhibitor, and specific antibodies to ROCK1 and ROCK2 proteins, which are two isoforms of ROCK, were used. Electrical field stimulation induced off-contraction and on-contraction in the presence of N<SUP>G</SUP>-nitro-L-arginine methylester, originating from the cholinergic nerve. Y-27632 inhibited both excitatory contractions in a concentration-dependent manner. Exogenous acetylcholine concentration-dependently induced two types of contractions: an initial contraction which occurred immediately after the addition of acetylcholine during short periods, and a sustained contraction which sluggishly continued after the initial contraction. Maximal initial and sustained contractions were reached at 10<SUP>-5</SUP> M acetylcholine. Y-27632 significantly inhibited both acetylcholine-induced contractions in a concentration-dependent manner. Western blot analysis revealed that acetylcholine maximally increased the level of phosphorylation in the 20 kDa regulatory light chain of myosin II (MLC<SUB>20</SUB>) at Ser<SUP>19</SUP> from 0.25 min to 1 min, and then declined after 2 min. The level changes of MLC<SUB>20</SUB> phosphorylation during the 5 min paralleled with those of acetylcholine-induced contractions. The expression of ROCK1 and ROCK2 in membrane fractions of muscle was increased by acetylcholine; more specifically, ROCK2 continually expressed up to 5 min. Taken together, ROCK may be involved in neural-evoked and acetylcholine-induced contraction via translocation to the membrane in feline esophageal smooth muscle.