GS-386이 단일 심근 세포의 Ca²⁺ 전류에 미치는 효과

박춘옥,장경재,김양미,한재희,홍성근,Park, Choon-ok,Chang, Kyeong-jae,Kim, Yang-mi,Haan, Jae-hee,Hong, Seong-geun 대한수의학회 1994 大韓獸醫學會誌 Vol.34 No.1

The effects of the novel compound GS-386 on the calcium current were investigated in rabbit atrial myocytes. The calcium current was recorded during various depolarizations of 200 ms duration from a holding potential of -40 mV using the whole cell patch clamp technique. The calcium current was activated from -30 mV, reached maximum amplitude at +10 mV and almost disappeared at +50 mV. Superfusion of GS-386 led to a reduction of the calcium current amplitude dose-dependently and $ED_{50}$ was $2.5{\times}10^{-7}M$. But the dependence of the calcium current on the membrane potential was not altered by GS-386. The inactivation of the calcium currents showed single exponential curves in both before and after application of GS-386. The inactivation time constants before and after application of GS-386 were almost the same(35 ms and 32.5 ms). The steady-state inactivation curve of the calcium current was not shifted by GS-386. The calcium currents both before and after application of GS-386 recovered completely in 1 sec and the recovery time constants were about 200 ms in both cases. From the above results it is concluded that the novel compound GS-386 has calcium antagonistic property decreasing the calcium current.

심장세포에서 세포내 Ca²⁺ 증가에 의해 활성화되는 Cl^- 통로의 특성과 역할

박춘옥,김양미,한재희,홍성근,Park, Choon-ok,Kim, Yang-mi,Haan, Jae-hee,Hong, Seong-geun 대한수의학회 1994 大韓獸醫學會誌 Vol.34 No.1

The inward tail current after a short depolarizing pulse has been known as Na-Ca exchange current activated by intracellular calcium which forms late plateau of the action potential in rabbit atrial myocytes. Chloride conductance which is also dependent upon calcium concentration has been reported as a possible tail current in many other excitable tissues. Thus, in order to investigate the exsitance of the calcium activated chloride current and its contribution to tail current, whole cell voltage clamp measurement has been made in single atrial cells of the rabbit. The current was recorded during repolarization following a brief 2 ms depolarizing pulse to +40mV from a holding potential of -70mV. When voltage-sensitive transient outward current was blocked by 2 mM 4-aminopyridine or replacement potassium with cesium, the tail current were abolished by ryanodine$(1{\mu}M)$ or diltiazem$(10{\mu}M)$ and turned out to be calcium dependent. The magnitudes of the tail currents were increased when intracellular chloride concentration was increased to 131 mM from 21 mM. The current was decreased by extracellular sodium reduction when intracellular chloride concentration was low(21 mM), but it was little affected by extracellular sodium reduction when intracellual chloride concentration was high(131 mM). The current-voltage relationship of the difference current before and after extracellular sodium reduction, shows an exponential voltage dependence with the largest magnitude of the current occurring at negative potentials, with is similar to current-voltage relationship at negative potentials, which is similar to current-voltage relationship of Na-Ca exchange current. The current was also decreased by $10{\mu}M$ niflumic acid and 1 mM bumetanide, which is well known anion channel blockers. The reversal potentials shifted according to changes in chloride concentration. The current-voltage relationships of the niflumic acid-sensitive currents in high and low concentration of chloride were well fitted to those predicted as chloride current. From the above results, it is concluded that calcium activated chloride component exists in the tail current with Na-Ca exchange current and it shows the reversal of tail current. Therefore it is thought that in the physiologic condition it leads to rapid end of action potential which inhibits calcium influx and it contributes to maintain the low intracellular calcium concentration with Na-Ca exchange mechanism.

Nitric Oxide Modulates Calcium Current in Cardiac Myocytes but not in Intact Atrial Tissues

박춘옥,강영진,이회영,장기철,Park, Choon-Ok,Kang, Young-Jin,Lee, Hoi-Young,Chang, Ki-Churl The Korean Society of Pharmacology 1995 대한약리학잡지 Vol.31 No.3

본 연구의 목적은 외부에서 nitric oxide (NO)를 투여 하였을때 심근 수축력, 심박동수의 변화 및 혈관 평활근에 대한 효과를 비교함으로서 NO에 대한 이들 장기의 민감도가 서로 같은지 또는 상이한지를 알아보고자 하였다. 본 실험에서는 PIANO 방법에 의한 근장력의 변화와 아울러 심근에서의 $Ca^{2+}$ current를 측정하였다. 랫트의 심방근에 대한 PIANO $(STZ,\;100\;{\mu}M)$는 심근수축력 및 심박동수에 전혀 변화를 주지 않았지만 혈관 평활근에서는 강한 이완 작용을 나타내었다. 한편, 8-Br-cGMP도 고농도 $(100\;{\mu}M)$에서만 심근 수축력을 억제하였다. 토끼의 심방근세포에서 Whole cell voltage patch clamp를 사용시 bradykinin, SNP, 8-Br-cGMP 및 PIANO는 $Ca^{2+}$ current를 억제하였다. 이러한 사실은 외부에서 공급되는 NO에 대한 심근과 혈관 평활근의 반응에는 민감도의 차이가 있음을 암시하며 더 나아가 심근의 경우에도 NO 반응에는 종 (species)간의 차이와 동일 종이라 하더라도 세포(cell)와 장기(tissue)에 차이가 있을 가능성을 제시하였다. The aim of the present study was to know whether exogenously administered nitric oxide (NO) may differently modulate muscle mechanics between heart and aorta. We used PIANO method to generate NO. In isolated rat atrial tissues, neither heart rate nor contractility was affected by PIANO $(STZ,\;30{\sim}100\;{\mu}M)$. Only high concentration $(100\;{\mu}M)$ of 8-bromo cyclic GMP slightly depressed cardiac contractility. However, the same concentrations of 8-Br cGMP and PIANO significantly relaxed the rat thoracic aorta contracted with phenylephrine $(0.1\;{\mu}M)$. In isolated rabbit cardiac atrial myocytes, the amplitude of calcium currents were decreased in the whole voltage range by the presence of streptozotocin, which was further potentiated by UV light. Calcium currents were also decreased in those preparations treated with bradykinin, nitroprusside and 8-Br cGMP. These findings suggest that exogenous NO may modulate calcium current in cardiac myocyte. However, it remains why this does not affect myocardial contractility and heart rate. We concluded that NO may differently regulate calcium signal between aorta and heart muscle.

심근세포 및 혈관 평활근에 대한 Nitric Oxide 작용의 민감성의 차이

박춘옥(Choon Ok Park),강영진(Young Jin Kang),이회영(Hoi Young Lee),장기철(Ki Churl Chang) 대한약리학회 1995 대한약리학잡지 Vol.31 No.3

본 연구의 목적은 외부에서 nitric oxide (NO)를 투여 하였을때 심근 수축력, 심박동수의 변화 및 혈관 평활근에 대한 효과를 비교함으로서 NO에 대한 이들 장기의 민감도가 서로 같은지 또는 상이한지를 알아보고자 하였다. 본 실험에서는 PIANO 방법에 의한 근장력의 변화와 아울러 심근에서의 Ca²⁺ current를 측정하였다. 랫트의 심방근에 대한 PIANO (STZ, 100μM)는 심근수축력 및 심박동수에 전혀 변화를 주지 않았지만 혈관 평활근에서는 강한 이완 작용을 나타내었다. 한편, 8-Br-cGMP도 고농도 (100μM)에서만 심근 수축력을 억제하였다. 토끼의 심방근세포에서 Whole cell voltage patch clamp를 사용시 bradykinin, SNP, 8-Br-cGMP 및 PIANO는 Ca²⁺ current를 억제하였다. 이러한 사실은 외부에서 공급되는 NO에 대한 심근과 혈관 평활근의 반응에는 민감도의 차이가 있음을 암시하며 더 나아가 심근의 경우에도 NO 반응에는 종 (species)간의 차이와 동일 종이라 하더라도 세포(cell)와 장기(tissue)에 차이가 있을 가능성을 제시하였다. The aim of the present study was to know whether exogenously administered nitric oxide (NO) may differently modulate muscle mechanics between heart and aorta. We used PIANO method to generate NO. In isolated rat atrial tissues, neither heart rate nor contractility was affected by PIANO (STZ, 30 ~ 100μM). Only high concentration (100μM) of 8-bromo cyclic GMP slightly depressed cardiac contractility. However, the same concentrations of 8-Br cGMP and PIANO significantly relaxed the rat thoracic aorta contracted with phenylephrine (0.1μM). In isolated rabbit cardiac atrial myocytes, the amplitude of calcium currents were decreased in the whole voltage range by the presence of streptozotocin, which was further potentiated by UV light. Calcium currents were also decreased in those preparations treated with bradykinin, nitroprusside and 8-Br cGMP. These findings suggest that exogenous NO may modulate calcium current in cardiac myocyte. However, it remains why this does not affect myocardial contractility and heart rate. We concluded that NO may differently regulate calcium signal between aorta and heart muscle.

사람의 Low Density Lipoprotein에 대한 녹차의 항산화 활성

박춘옥(Park Chun Ok),진성현(Jin Seung Heun),류병호(Ryu Beung Ho) 한국식품과학회 1996 한국식품과학회지 Vol.28 No.5

수축과 활동전압의 Staircase 현상에 대한 Ca-전류 및 세포내 Ca²⁺ 농도 변화의 영향

박춘옥(Park, Choon-Ok),서인석(So, In-Suk),호원경(Ho, Won-Kyung),엄융의(Earm, Yung-E),김우겸(Kim, Woo-Gyeum) 대한생리학회 1989 대한생리학회지 Vol.23 No.2

It well known that the magnitude of contraction and the shape of action potential depend upon the stimulation frequency and the duration of resting period (positive and negative staircase). Although the underlying mechanism of the staircase phenomenon is not fully understood, it has been suggested that staircase could be related to the intracllular Ca²⁺ concentration. In order to elucidate the role of intracellular Ca²⁺ on the contraction and action potential staircases, we examined the effects of 1 mM 4-aminopyridine (4-AP), 0.5 uM verapamil, 1 uM ryanodine, or reduction of extracellular Na concentration to 30% (substituted by equimolar Li⁺) in small atrial strips of the rabbit (3 X 10 mm). The results obitained were as follows; 1) When the stimulation frequency was increased from 0.1 Hz to 2 Hz, positive staircase of the contraction and elevation of plateau level in action potential were found in control and the conditions of Na reduction and treatments of 4-AP, verapamil and ryanodine. 2) When stimulation frequency returned to 0.1 Hz from 1 min rest just after 2 Hz stimulation fer 1 min, the magnitudes of initial few contractions were larger than that of steady state contraction (post-rest potentiation) except, ryanodine or Na-reduction groups. 3) Negative staircase of contraction was developed in control and 4-AP group at post-rest 0.1 Hz stimulation and the plateau level of the action potential was decreased at the same time. But the reduction of contraction or the plateau level was much smaller in 4-AP group and than in control. From the above results it can be concluded that contraction and action potential staircase is dependent upon transmembrane Ca²⁺-current and Ca²⁺release from the SR.

수정에 의한 Mouse egg의 세포막전류 변화

홍성근,박춘옥,한재희,김익현,하대식,권종국,Hong, Seong-geun,Park, Choon-ok,Han, Jae-hee,Kim, Ik-hyun,Ha, Dae-sik,Kwun, Jong-kuk 대한수의학회 1992 大韓獸醫學會誌 Vol.32 No.2

Changes in the both inward current and conductance of membrane by the fertilization were observed using the one microelectrode voltage clamp(or switch clamp) technique. Unfertilized eggs and both 1- and 2-cell stage eggs after fertilization were donated from the superovulated mouse (ICR, more than 6 weeks old) treated with PMSG(pregnant mare serum gonadotropin, Sigma) and HCG(human chorionic gonadotropin, Sigma) and naturally mated ones, respectively in this experiment. Membrane potential was held at -90mV and the voltage step was applied from -80mV to 50mV with interval of 10mV or 20mV for 300ms. since both of amplitudes and time courses in the membrane currents were various according to the states of cells and clamping condition, results were presented by their $averages{\pm}SEM$(standard mean error)and ratios or percentages. Inward currents began to appear in response to the step depolarization from -60mV and reached its maximum at -50mV. However, since the potential was not clamped evenly during the voltage step, current-voltage(I-V) relationship might be positively shifted 10 or 20mV. From the steady-state currents plotted in the I-V curve, outward rectification was markedly observed. Peak inward currents$(i_{in})$ at -50mV were $-0.62{\pm}0.23nA$(n=4),$-0.52{\pm}0.25nA$(n=5) and $-0.37{\pm}0.25nA$(n=6), in the 1-cell stage, 2-cell stage fertilized eggs and in the unfertilized eggs, respectively. Pure inward current (difference between steady-state and peak, $i_{in. pure}$) were $-1.01{\pm}0.23nA$, $-0.69{\pm}0.43nA$ and $-0.68{\pm}0.29nA$, respectively in the 1-cell stage fertilized eggs, unfertilized eggs and 2-cell stage fertilized eggs. These results suggested that the outward current in fertilized eggs of 2-cell stage was more increased than those in the unfertilized eggs. Pure inward currents in the all stages of eggs showed a similar fashion in the I-V relationship from -50mV to 50mV and reversal potential at 50mV. Time constant of inactivation$({\tau})$ in the inward current was decreased as the membrane potential was depolarized in the unfertilized and 2-cell stage eggs but in the 1-cell stage eggs t was not likely to be affected significantly. Slope conductances were 14.2nS, 8.9n5 and 7.7nS in the 1-cell, 2-cell stage fertilized eggs and the unfertilized eggs, respectively. Membranes between two cells within a zona pellucida seem to be electrical-connected in the 2-cell stage eggs from the observation made in the analysis for the electronic spread and decay to the current stimuli. Both of inward current and membrane conductance were increased after fertilization in the mouse eggs. Inward current seems to be carried by the same ion or through the same channels up to the 2-cell stage and ion that carried inward current was thought to play important function after fertilization in the mouse eggs.

Effects of Extracellular $Ca^{2+}$ and $Ca^{2+}$-Antagonists on Endothelium-Dependent Relaxation in Rabbit Aorta

서석효,구용숙,박춘옥,황상익,김기환,Suh, Suk-Hyo,Goo, Yong-Sook,Park, Choon-Ok,Hwang, Sang-Ik,Kim, Ki-Whan The Korean Physiological Society 1990 대한생리학회지 Vol.24 No.1

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 effects of extracellular $Ca^{2+}$ and various $Ca^{2+}$ antagonists on endothelium-dependent relaxation to acetylcholine were studied in the isolated rabbit thoracic aorta in order to elucidate the control mechanism of endothelium derived relaxing factor (EDRF) release. Endothelium was removed from aortic strips by gentle rubbing with cotton ball. The effect of hemoglobin on basal tension was also observed with hemolysate. The results obtained were as follows: 1) Endothelium-dependent relaxation (EDR) to acetylcholine (ACh) showed biphasic pattern; the initial rapid relaxation phase and the late slow relaxation phase. 2) With the depletion of the extracellular $Ca^{2+}$, EDR was gradually suppressed, especially the late slow relaxation. 3) Verapamil, nifedipine, $Mn^{2+}$ and $Cd^{2+}$ had not any effect on EDR, while $La^{3+}$ and $Co^{2+}$ suppressed EDR completely. 4) The resting tension of the strips with rubbed endothelium was not altered by the addition of hemoglobin. That of the strips with intact endothelium, however, was enhanced and EDR to ACh was completely blocked From these results, we suggest that extracellular $Ca^{2+}$ is necessary for ACh-induced slow relaxation while $Ca^{2+}$ antagonists have not any effect on EDR.

A Comparative Study on the Electrophysiological Properties of Medial and Lateral Spinoreticular Tract Cells in Cats

이석호,전제열,박춘옥,구용숙,김전,성호경,Lee, Suk-Ho,Jun, Jae-Yeol,Park, Choon-Ok,Goo, Yong-Sook,Kim, Jun,Sung, Ho-Kyung The Korean Physiological Society 1990 대한생리학회지 Vol.24 No.1

Vasomotor area로 알려져 있던 외측연수망상체는 최근 rostral ventrolateral medulla로 불리 면서 sympathoexcitatory neuron이 집중적으로 분포하는 연수내 부위로서, 체성교감신경반사에 중요한 역할을 하는 것으로 알려지고 있어 이 부위로의 입력정보가 그동안 많은 연구가 되어온 내측 척수망상로 세포와 어떤 차이를 보이는가를 규명하고자 하였다. 1) Medial SRT 세포는 34 cell중 약 60%가 동측으로 향하였으나, lateral SRT cell의 경우 47 cell의 약 60%가 반대측으로 향하였다. 2) 각 군의 세포를 말초자극에 대한 반응성에 따라, LT cell, Deep cell, HT cell및 WDR cell로 나누었으며 유해자극을 전달하는 세포를 HT와 WDR cell이라 하고, 무해자극을 전달하는 세포와 유해자극을 전달하는 세포의 비율로 볼 때, 다른 부위에 비하여 rostral VLM에 유해자극정보가 비교적 많이 전달됨을 볼 수 있었다. 3) 평균 전도 속도는 각 군간에 유의한 차이가 없었으나, Deep cell은 HT cell보다 유의하게 빠른 전도속도를 보였다. 4) Medial SRT 세포는 척수 회백질 내에서 Rexed laminae VII및 VIII에 주로 분포하며, LRN projecting SRT cell의 경우엔 전 lamina에 걸쳐 고루 분포하였다. 이상의 결과는 내측 및 외측 망상체간의 말초 입력 정보의 차이를 보여주고 있으며, 연수 망상체의 세 부위중 특히 rostral ventrolateral medulla로 많은 동통정보가 입력되고 있어 RVLM의 기능중 하나인 체성교감반사의 통합과 연관된 역할을 하리라 기대된다. Antidromically activated spinoreticular tract (SRT) cell units in the lumbosacral enlargement of ${\alpha}-chloralose$ anesthetized cats were classified as medial and lateral SRT units according to the location of their axonal termination. Identified SRT units were tested fer antidromic conduction velocity, laterality of their axonal projection, the location in spinal gray, peripheral receptive field, the response pattern to graded mechanichal stimulation and the responsiveness to $A{\delta}$ and C volley of the peripheral nerve. 1) The 59% of 34 medial SRT units were recorded in ipsilateral side to the antidromic stimulation site, but 60% of the 47 lateral SRT units projected to contralateral side. 2) Most of the medial SRT cells and rostral ventrolateral medulla (RVLM)-projecting lateral SRT cells were recorded in lamina VII & VIII. The LRN (lateral reticular nucleus)-projecting SRT cells, however, distributed through all the laminae except superficial ones (I & II). 3) The identified SRT units were classified as low theshold (LT), deep, high threshold (HT), wide dynamic range (WDR) cells, based on the response patterns to graded mechanical stimuli. The proportion of SRT units which receive noxious input was 37.5%, 25% and 75% in the medial, LRN-projecting and RVLM SRT group, respectively. 4) There was no significant difference in the mean conduction velocities between the 3 groups. But the deep cells had significantly higher velocity than that of the HT cells. The above results show that the peripheral inputs to the SRT units are different in the 3 groups: medial, LRN & RVLM SRT group. Especially in case of the SRT cells projecting to RVLM which is a probable candidate fur the integration center of various pressor reflexes such as somatosympathetic reflex, the noxious informations occupy higher proportion of input to them than in other groups. Therefore the noxious information transmitted through the lateral SRT destined for RVLM is expected to play a role in somatosymapthetic reflex.

춘천지역 안개의 화학적 조성 ( 제1보 ) : 안개 포집기의 개발

김만구,정일권,임양석,박춘옥 ( Man Goo Kim,Il Keun Jung,Yang Suck Lim,Chun Oak Park ) 대한환경공학회 1993 환경종합 학술대회 Vol.- No.-