소(牛) 식도구 평활근의 Adrenergic receptor 존재부위에 관한 연구

강동묵,조제열,박전홍,양일석,Kang, Tong-mook,Cho, Je-yoel,Park, Jun-hong,Yang, Il-suk 대한수의학회 1993 大韓獸醫學會誌 Vol.33 No.4

The preliminary studies on the localization of adrenoceptors were performed on smooth muscle strips of bovine esophageal groove. The mechanical activity of the muscle strip was recorded isometrically in vitro.w In the bottom circular muscle strips. the excitatory ${\alpha}-adrenergic$ responses were not blocked by tetrodotoxin$(2.1{\times}10^{-6}M)$ and denervation which was carried by cold storage of strips for 48 hrs in Tyrode's solution at $5-6{^{\circ}C}$ without oxygen supply. These excitatory ${\alpha}-adrenergic$ responses were partially blocked by atropine. In the lip longitudinal muscle strips, the inhibitory${\beta}-adrenergic$ responses were not blocked by pretreatment of tetrodotoxin and atropine. The results suggest that ${\beta}-adrenergic$ receptors mediating relaxations are located on the postsynaptic smooth muscle cells, whereas ${\beta}-adrenergic$ receptors mediating contractions are located both in the smooth muscle cells and in the cholinergic neurones.

소(우(牛)) 식도구 윤상근의 비아드레날린 비콜린성 이완 및 수축

강동묵,한호재,양일석,Kang, Tong-mook,Han, Ho-jae,Yang, Il-suk 대한수의학회 1995 大韓獸醫學會誌 Vol.35 No.2

To characterize non-adrenergic non-cholinergic(NANC) nerve mediated contractile responses in circular smooth muscle of bovine reticular groove, we investigated NANC relaxation and contraction induced by electric field stimulation to enteric nerves. In the presence of atropine($1{\mu}M$) and guanethidine($50{\mu}M$), electric field stimulation at frequency of 1 to 16Hz(square pulses, 0.5ms duration, 70V) evoked clear-cut relaxations through stimulations. Transient 'rebound contraction' was occured when the stimulus was switched off. All of the responses (relaxation and rebound contraction) were dose-dependently blocked by Nw-nitro-$_{\small{L}}$-arginine methyl ester(L-NAME), an inhibitor of nitric oxide synthesis, and methylene blue, and inhibitor of soluble guanylate cyclase. Tetraethyl ammonium(TEA), a potassium channel blocker, did not block the NANC relaxations.

Nitric oxide에 의한 수퇘지 음경후인근의 비아드레날린 비콜린 동작성 이완 II. 비아드레날린 비콜린성 신경의 전장자극과 S-nitrosothiols에 의한 돼지 음경후인근의 이완 효과 비교

문규환,김태완,강동묵,이완,양일석,Mun, Kyu-whan,Kim, Tae-wan,Kang, Tong-mook,Lee, Wan,Yang, Il-suk 대한수의학회 1995 大韓獸醫學會誌 Vol.35 No.3

As S-nitrosothiols were proposed as nitrergic carriers in vascular and nonvascular smooth muscle, we have investigated the relaxant properties of several S-nitrosothiols in the porcine retractor penis(PRP) muscle and compared them with the effects of exogenously added NO, electrical field stimulation(EFS) of NANC nerves and sodium nitroprusside(SNP). Also the influences of oxyhemoglobin and hydroquinone on the relaxant responses were investigated. In addition, effects of NO on membrane potentials and its involvement in the generation of inhibitory junction potential(IJP) were investigated with conventional intracellular microelectrode technique. The results were summerized as follows. 1. Frequency-dependent relaxations of PRP muscle were induced by EFS to NANC nerve. Tetrodotoxin($1{\times}10^{-6}M$) abolished the relaxations of PRP muscle induced by EFS, and L-NAME(($2{\times}10^{-5}M$) and methylene blue($4{\times}10^{-5}M$) inhibited the relaxations. L-NAME-induced inhibition of the relaxations was reversed by L-arginine($1{\times}10^{-3}M$), but not by D-arginine. 2. Exogenous NO($1{\times}10^{-5}-1{\times}10^{-4}M$), sodium nitroprusside(($1{\times}10^{-7}-1{\times}10^{-4}M$) induced dose-dependent relaxations of PRP muscle. All S-nitrosothiols($1{\times}10^{-7}-1{\times}10^{-4}M$) tested relaxed the PRP muscle in dose-dependent manner and the potency order was SNAP>GSNO>CysNO>SNAC. 3. Oxyhemoglobin($5{\times}10^{-5}M$) blocked the relaxation induced by exogenous NO and inhibited EFS-, S-nitrosothiols-, and SNP-induced relaxation. 4. Hydroquinone($1{\times}10^{-4}M$) also abolished the relaxations induced by exogenous NO, and reduced the relaxations induced by S-nitrosothiols, but did not affect EFS- and SNP-induced relaxations. 5. SNP($2{\times}10^{-6}-5{\times}10^{-6}M$) relaxed muscle strips but the membrane potentials were not affected. 6. EFS with several pulses(1ms, 2Hz, 80V) produced an inhibitory junction potential(IJP) with muscle relaxation. They were abolished by TTX($2{\times}10^{-6}M$). $N^G$-nitro-$_{\small{L}}$-arginine(L-NNA, $2{\times}10^{-5}M$) abolished the muscle relaxation, but had no effect on IJP.

소 음경후인근의 Nitric oxide(NO) 매개성 이완

양일석,장희정,강동묵,이장헌,Yang, Il-suk,Chang, Hee-jung,Kang, Tong-mook,Lee, Jang-hern 대한수의학회 1996 大韓獸醫學會誌 Vol.36 No.3

This study was designed to examine the mechanism of penile erection in adult bull by analyzing the responses of bovine proximal retractor penile muscle strips(BRP) to electtical field stimulation(EFS), exogenous nitric oxide(NO), NO synthesis precursor(L-arginine), NO synthase inhibitors(L-NAME, L-NMMA), guanylate cyclase inhibitor(methylene blue) and nonspecific potassium channel blocker(tetraethylammonium, TEA) treatments. Isometric tension of BRP was measured using physiograph. Results were summarized as follows: 1. EFS of nonadrenergic noncholinrgic(NANC) nerve in BRP produced frequency-dependent inhibitory responses to the contraction induced by co-treatment of epinephrine, guanethidine and atropine. The inhibitory responses to EFS were blocked by tetrodotoxin(TTX, $1{\mu}M$). 2. Treatment of L-NAME ($10,\;20{\mu}M$) inhibited the relaxation to EFS whereas L-NMMA ($100{\mu}M$) had no effect. 3. Treatment of NO($20,\;40{\mu}M$; as an acidified solution of $NaNO_2$) induced concentration-dependent relaxation whereas preincubation of TTX($1{\mu}M$) and L-NAME($20{\mu}M$) had no effect on the relaxation response. 4. L-arginine treatment(10mM) blocked the inhibitory effect of L-NAME($20{\mu}M$). 5. Pretreatment of methylene blue($40{\mu}M$) reduced the NANC-induced relaxation of BRP. 6. Tetraethylammonium(TEA, 80mM) reduced NANC relaxation. These results suggest that NO may act as a NANC neurotransmitter in BRP and the effects might be mediated by cGMP and potassium channel.

Nitric oxide에 의한 수퇘지 음경후인근의 비아드레날린 비콜린 동작성 이완 I. 돼지 음경후인근의 비아드레날린 비콜린성 이완을 매개하는 신경전달물질 : nitric oxide와 vasoactive intestinal polypeptide

문규환,김점용,김태완,강동묵,양일석,Mun, Kyu-whan,Kim, Jeum-yong,Kim, Tae-wan,Kang, Tong-mook,Yang, Il-suk 대한수의학회 1995 大韓獸醫學會誌 Vol.35 No.3

This study was carried out to characterize nonadrenergic, noncholinergic(NANC) relaxation of porcine retractor penis(PRP) muscle induced by electrical field stimulation(EFS) and to investigate the actions of niric oxide(NO) and vasoactive intestinal polypeptide(VIP) as candidates for NANC neurotransmitters. Biphasic relaxations of PRP muscle were induced by EFS to NANC nerve. Rapid-phase relaxation was observed at low frequency(0.5-16Hz) and slow-phase relaxation followed during high frequency(8-60Hz). Both relaxations were frequency-dependent and TTX($1{\times}10^{-6}M$)-sensitive. L-NAME($2{\times}10^{-5}M$) inhibited the rapid-phase relaxation, but not the slow-phase relaxation. The inhibition of the rapid-phase relaxation with L-NAME was reversed by L-arginine ($1{\times}10^{-3}M$) but not by D-arginine($1{\times}10^{-3}M$). Methylene blue($4{\times}10^{-5}M$) reduced the rapid-phase relaxation. Exogenous No(ExoNO, $1{\times}10^{-5}-1{\times}10^{-4}M$) induced dose-dependent relaxations of PRP muscle. Oxyhemoglobin($5{\times}1^{-5}M$) blocked the relaxation induced by ExoNO and inhibited EFS-induced relaxation. Hydroquinone($1{\times}10^{-4}M$) also abolished the relaxation induced by ExoNO, but did not affect EFS-induced relaxation. L-NAME resistant slow-phase relaxation to EFS was inhibited by ${\alpha}$-chymotrypsin(2.5 U/ml). Both methylene blue($4{\times}10^{-5}M$) and Nethylmaleimide($1{\times}10^{-4}M$) reduced the slow-phase relaxation by EFS. [4-Cl-D-$Phe^6$, $Leu^{17}$]-VIP($3{\times}10^{-6}M$) inhibited the slow-phase relaxation by EFS. External applications of VIP ($1{\times}10^{-7}M$) caused relaxations that were simillar to the L-NAME resistant slow-phase relaxations induced by EFS, and relaxant effects of exogenous VIP were blocked by ${\alpha}$-chymotrypsin(2.5 U/ml).

기니피그 담낭 평활근에서 무스카린 수용체 아형의 특성

이종균(Jong Kyun Lee),이풍렬(Poong Lyul Rhee),강동묵(Tong Mook Kang),김재준(Jae Jun Kim),고광철(Kwang Cheol Koh),백승운(Seung Woon Paik),이종철(Jong Chul Rhee) 대한소화기학회 1998 대한소화기학회지 Vol.30 No.1

N/A Background/Aims: Acetylcholine(Ach) is the most irnportant rnediator in vagal-induced and cholecystokinin-induced gallbladder contraction. There are several rnuscarinic receptor subtypes characterized by pharmacologic study. The majority of smooth muscles from many species exhibit heterogenous populations of muscarinic receptors and different subtypes activate different signal transduction pathways. Therefore, it is important and essential to characterize the muscarinic receptor subtypes for understanding the mechanism of muscle contraction in a certain tissue. The aim of the present study was to characterize the muscarinic receptor subtypes in guinea pig gallbladder smooth muscle using relatively selective receptor subtype antagonists. Methods: Gallbladder muscle strips were prepared and mounted in an organ bath. Isometric contractions were recorded on a polygraph. 50% inhibitory concentrations(Icqp) and pA (-log KB) values of pirenze- pine, methoctramine, and 4-DAMP(M1, M2, and M3 antagonist, respectively) were compared on Ach-induced gallbladder muscle contraction. Results: Icsp of pirenzepine, methoctramine, and 4-DAMP on 1pM Ach-induced contraction were 7.8 > 10 'M, 2.'7 x 10 M, and 3,7 x 10 M, respectively. pA, values of pirenzepine, methoctramine, and 4-DAMP were 6.8, 5.0 and 11.4, respectively. Conclusions: M3 subtype may play the most important role in Ach-induced contraction of guinea pig gallbladder muscle. (Korean J Gastroenterol 1997; 30:90 - 97)

사람 위 평활근 세포 내에서 카바콜에 의한 칼슘 농도 증가의 기전

문원(Won Moon),김영호(Young Ho Kim),박동일(Dong Il Park),이풍렬(Poong Lyul Rhee),김재준(Jae J . Kim),고광철(Kwang Cheol Koh),이종철(Jong Chul Rhee),최규완(Kyoo Wan Choi),강동묵(Tong Mook Kang) 대한내과학회 2001 대한내과학회지 Vol.60 No.5

N/A Background : The contraction of smooth muscle depends on an increase in the concentration of intracellular calcium ion and the source of this increase to various stimuli is different according to organs or species. Nevertheless, there have been only a few studies on human stomach smooth muscle. This study was designed to identify the source of the calcium utilized in the muscle contraction induced by carbachol, which is an important factor among various stimuli. Methods : After the administration of carbachol with various conditions in cultured human stomach smooth muscle cells, fura-2-acetoxymethyl ester was used to measure the increase in the intracellular calcium concentration. Results : (1) The carbachol-induced increase in the intracellular calcium concentration was not attenuated after removal of extracellular calcium. (2) Carbachol induced a small increase in the intracellular calcium concentration even after the depletion of intracellular calcium store. (3) Repeated histamine administration blocked the carbachol-induced increase in the intracellular calcium concentration in calcium-free extracellular solution. Conclusion : The main source of calcium utilized in human stomach smooth muscle contraction by carbachol is intracellular calcium store, particularly inositol triphosphate(IP3) - sensitive calcium stores. However, extracellular calcium also contributes to the carbachol-induced increase in the intracellular calcium concentration. (Korean J Med 60:432-438, 2001)

모틸린에 의한 사람 위 평활근의 수축 기전에 관한 연구

심상군(Sang Goon Shim),이종철(Jong Chul Rhee),이풍렬(Poong Lyul Rhee),최규완(Kyoo Wan Choi),전성국(Sung Kook Jeon),강동묵(Tong Mook Kang),엄대용(Dae Yong Uhm),이종석(Jong Seok Lee),성인경(In Kyung Sung),김현서(Hyun Seo Kim) 대한소화기학회 2002 대한소화기학회지 Vol.39 No.1

Background/Aims: Motilin is an intestinal peptide that stimulates the contraction of gut smooth muscle. A discrepancy exists between the in vivo (neurally mediated) and in vitro (direct action on a smooth muscle receptor) mechanisms of motilin action in many species. We investigated in vitro mechanisms of motilin action on human gastric smooth muscle. Methods: Antral cirular muscle strips of the surgical tissue obtained during gastrectomy, were used to measure contractile force and electrical activity. Dispersed muscle cells were used to measure L-type Ca2+ current and electrical activity. Results: Motilin of 1-100nM contracted smooth muscle in a concentration-dependent manner. Motilin-induced contractions were unaffected by tetrodotoxin or atropine treatment. Nifedipine or Ca2+-free bath solution blocked motilin (10nM)-induced contractions. Low concentration of motilin (1nM) resulted in an increase in acetylcholine (0.1~100M)-induced contractions. By patch clamp recording technique, motilin (1 or 10nM) did not modify the L-type Ca2+ current, but motilin-induced membrane depolarization was detected. Erythromycin also contracted smooth muscle with membrane depolarization but verapamil inhibited the contraction. Conclusions: These results suggest that motilin contracts smooth muscle through a direct action on smooth muscle receptor and Ca2+ influx through the L-type Ca2+ channel, which is due to membrane depolarization, also mediates motilin-induced contractions. (Korean J Gastroenterol 2002;39:4-12)

아세틸콜린과 콜레시스토키닌에 의한 사람 담낭 수축시 이용되는 칼슘의 근원

강동묵,이종균,이풍렬,이종철,윤용범,엄대용 대한소화기학회 1998 대한소화기학회지 Vol.31 No.6

Background/Aims: Gallbladder contraction regulates the bile flow during digestive period and pevents stasis of the lithogenic bile. In addition, abnormal gallbladder motility may cause recurtent biliary pain or biliary dyskinesia. Acetylcholine (ACh) and cholecystokinin (CCK) are major neurohormonal mediators in gallbladder contraction. In addition, an increase in cytosolic calcium concentration is a common event resulting in smooth muscle contraction. This study was designed to identify the sources of calcium utilized in gallbladder smooth muscle contraction mediated by ACh and CCK in human. Methods: By measuring the contractile forces of the muscle strips and the intracellular free Ca^(2+) conentration, Ca^(2+) sources utilized in muscle contraction were estimated. Results: The ACh-induced contractile response was accentuated after a twofold increment of the extracellular calcium concentration. The contractile response to ACh was markedly blocked by verapamil (10 μM) and was significantly potentiated by Bay-K 8644 (1 μM). Preventing Ca^(2+)release from the internal stores by strontium (4 mM) reduced the contractile response to ACh and CCK. The inhibitory effect was greater in the response to CCK than in the response to ACh Depletion of IP3-sensitive calcium stores by a high concentration of hista mine (50 μM) significantly inhibited the contractile response to CCK. The intfacellular caleium concentration was increased slowly on ACh-induced contraction but rapidly on CCK-induced contraction which was similar to the behavior of caffeine. Conclusions: These results suggest that ACh utilizes both intracellular and extracellular calcium, mainly extracellular calcium, whereas CCK utilizes calcium from intracellular calcium stores.

기니피그 담낭 평활근에서 무스카린 수용체 아형의 특성

이종균,이풍렬,강동묵,김재준,고광철,백승운,이종철 大韓消化器病學會 1997 大韓消化器病學會誌 Vol.30 No.1