Ovalbumin으로 감작된 기니픽 기관평활근의 수축을 이용한 천식치료제 효능검색

제현동,허인회 중앙대학교 약학연구소 1996 약학 논총 Vol.10 No.-

Direct bronchial dilators, antihistamines, inhibitors of mast cell degranulations or glucocortioids are major therapeutics in current use against allergic asthma. However, every one of them has certain degree of drawbacks spanning from lack of full efficacy to frank side effects, restricting the long term use. There have been continuous efforts to develop new antiasthmatic agents with high efficacy and low side effects. A reliable, fast, and economic method of efficacy screening is crucial to the success of the new drug development. There have been several reports of animal models of allergic asthma, in which clinical hallmarks of asthma such as hypersensitive bronchial constriction with smooth muscle hyperreactivity and eosinophilia in alveoli and bronchial tissue were duplicated. In the current study, a fast, reliable, and economic method for the screening new antiasthmatic agents was established based on these animal models. Hartley guinea pig was actively sensitized to ovalbumin by subcutaneous injection of ovalbumin/aluminum hydroxide mixture on day O and 14. Three weeks after the initial injection, tracheal smooth muscle strip was excised and suspended in a tissue chamber. The chamber was superfused with Krebs- Ringer bicarbonate buffer (pH 7.4) at 37 ℃ and gassed with 95% O_2, 5% CO_2 mixture. Contractions of the muscle was monitored with a isometric tension transducer. Ovalbumin (1 ug/ml) introduced into the chamber caused a sustained contraction of the tracheal smooth muscle strips from sensitized guinea pigs, while the muscle from non-sensitized control animal did not respond. Histamine induced same degree of contractions of tracheal smooth muscle either sensitized or control animals. Three stips can be obtained from each animal, and three or more dose points were obtained on each strip either with wash between experiments or in a cumulative dosage manner. The ovalbumin- induced contraction was relaxed, in a dose-dependent manner by salbutamol, isoproterenol, theophylline or by atropine, implying direct relaxing effects of these agents, Pretreatment of the muscle strip with cromoglycate, pyrilamine, or thoephylline diminished ovalbumin-induced contractions, implying preventive effects of these agents. High potencies of pyrilamine (EC_50 = 1.00×10^-7M) and cromoglycate (EC_50 = 2.14×10^10-5M) effects suggest that ovalbumin-induced contraction is mainly due to the released histamine from mast cells, which is compatible to the known pathophysiology of allergic asthma. Using this screening model, twenty test drugs prepared from herbal resources were screened. Three drugs showed both of relaxing and preventive effects: and one showed only preventive effect. Especially, these drugs showed complete prevention when given before ovalbumine, while cromoglycate reached only 60% prevention. In conclusion, the current model of ex vivo tracheal smooth muscle contraction from sensitized guinea pig was proved reliable, fast and economic screening methods for antiasthmatic agents under development. Some of test drugs screened so far proved effective, justifying further purification and decimalization.

Endothelium-Independent Effect of Fisetin on the Agonist-Induced Regulation of Vascular Contractility

제현동,손의동,나현오 한국응용약물학회 2016 Biomolecules & Therapeutics(구 응용약물학회지) Vol.24 No.1

Fisetin, a natural flavonoid found in a variety of vegetables and fruits, has been shown to possess many biological functions. The present study was undertaken to investigate the influence of fisetin on vascular smooth muscle contractility and to determine the mechanism involved. Denuded aortic rings from male rats were used and isometric contractions were recorded and combined with molecular experiments. Fisetin significantly relaxed fluoride-, thromboxane A2- or phorbol ester-induced vascular contraction suggesting as a possible anti-hypertensive on the agonist-induced vascular contraction regardless of endothelial nitric oxide synthesis. Furthermore, fisetin significantly inhibited fluoride-induced increases in pMYPT1 levels and phorbol ester-induced increases in pERK1/2 levels suggesting the mechanism involving the inhibition of Rho-kinase activity and the subsequent phosphorylation of MYPT1 and MEK activity and the subsequent phosphorylation of ERK1/2. This study provides evidence regarding the mechanism underlying the relaxation effect of fisetin on agonist-induced vascular contraction regardless of endothelial function.

The Inhibitory Effect of Rosiglitazone on Agonist-Induced or Spontaneous Regulation of Contractility

제현동,Sun Young Park,Amy L. Barber,손의동 대한약학회 2007 Archives of Pharmacal Research Vol.30 No.4

The present study was undertaken to determine whether rosiglitazone treatment influences on the agonist-induced or spontaneous regulation of vascular smooth muscle contraction and, if so, to investigate the related mechanism. Stimulants were directly added without any preanesthetic stress or spontaneous vasoconstriction was induced by preanesthetic physical stress where rat aortic ring preparations isolated from rat exposed to preanesthetic stress such as pinch or prick for 30 min were mounted in organ baths and then exposed to contractile agents. Previously and subchronically ingested rosiglitazone decreased Rho-kinase activating agonistinduced contraction but not depolarization- or α adrenergic agonist-induced contraction. Moreover, preanesthetic stress induced the stress-induced spontaneous contraction and previously and subchronically ingested rosiglitazone abolished the stress-induced spontaneous contraction. In conclusion, this study provides the evidence and possible related mechanism concerning the vasorelaxing effect of an antidiabetic rosiglitazone as an antihypertensive on the agonist-induced contraction or stress-induced spontaneous vasoconstriction in rat aortic rings regardless of endothelial function.

브로콜리 유래 Sulforaphane의 혈관 수축성 조절 효과

제현동 대한약학회 2014 약학회지 Vol.58 No.4

The present study was undertaken to investigate the influence of sulforaphane on vascular smooth muscle con- tractility and to determine the mechanism involved. We hypothesized that sulforaphane, the primary ingredient of broccoli of cruciferous vegetables, plays a role in vascular relaxation through inhibition of Rho-kinase in rat aortae. Intact of denuded arterial rings from male Sprague-Dawley rats were used and isometric tensions were recorded using a computerized data acquisition system. Interestingly, sulforaphane significantly inhibited fluoride, phorbol ester or thromboxane A2 mimetic-induced contraction in denuded muscles suggesting that additional pathways different from endothelial nitric oxide synthesis such as inhibition of Rho-kinase or MEK might be involved in the vasorelaxation. Furthermore, sulforaphane inhibited thromboxane A2-induced increases in pERK1/2 levels suggesting the mechanism including inhibition of thromboxane A2-induced increases in ERK1/2 phosphorylation. This study provides evidence that sulforaphane induces vascular relaxation through inhibition of Rho-kinase or MEK in rat aortae.

지치 유래 naphthoquinone을 전처치한 생쥐에서 우울 및 불안 조절 효과

제현동,민영실 중소기업융합학회 2020 융합정보논문지 Vol.10 No.7

본 연구는 불안, 불면증, 우울증 또는 삶의 질에 대하여 지치(Lithospermum erythrorhizon) 유래 naphthoquinone, 즉 shikonin의 영향을 고찰하고, 확실히 밝혀지지 않은 메카니즘을 규명하고자한다. 우리는 지치의 주요성분인 naphthoquinone이 스트레스에 의한 수면장애, 강제수영에 의한 우울, 미로에 의해 유발된 불 안현상 등의 조절에 영향을 미칠 것이라고 가설을 세웠다. 수컷 쥐를 이용하여 부동 혹은 수영시간, 수면시간, open arms으로 지속시간 및 진입빈도를 측정하고 기록하였다. Naphthoquinone(10, 30 & 100mg/kg)의 투여군에서 GABAA receptor의 활성으로 일어나는 barbiturate-유도 수면을 증가시켰다. 미로에서 open arms 상태로 지속 되는 시간이 증가되고, 강제수영에서 부동시간이 줄었다. 결과적으로 naphthoquinone은 불안을 완화, 수면과 항 우울경향이 있고, 불안, 불면과 우울증에서 효과적인 치료효과가 있었다. This study was undertaken to investigate the influence and related mechanisms that have yet to be clearly demonstrated of Lithospermum erythrorhizon derived-naphthoquinone (shikonin) on the anxiety, insomnia, depression in rats. We hypothesized that naphthoquinone, the primary ingredient of Lithospermum erythrorhizon, plays a role in the modulation of insomnia evoked by stress, depression evoked by forced swimming or anxiety evoked by elevated plus maze. Male ICR (Institute of Cancer Research) mice were used and the immobility or swimming time, the duration of sleep, the duration and entry frequency into open arms were measured and recorded. The administration of naphthoquinone (10, 30 and 100 mg/kg) potentiated barbiturate-induced sleep suggesting the activation of GABAA receptor. It also potentiated the time spent in open arms of the maze and decreased the immobility time in forced swimming. In conclusion, naphthoquinone has anxiolytic, hypnotic and anti-depressant properties and is a potential therapeutic for anxiety, insomnia and depression.

SM22α Is Required for Agonist-induced Regulation of Contractility: Evidence from SM22α Knockout Mice

제현동,손의동 한국분자세포생물학회 2007 Molecules and cells Vol.23 No.2

혈관 수축성에 대한 phenylephrine, isoprenaline 및 prazosin의 융합성 조절 효과

제현동,민영실 중소기업융합학회 2022 융합정보논문지 Vol.12 No.4

심혈관계 활성이 예측되는 phenylephrine, isoprenaline, prazosin의 단독 및 병용 투여에서 혈관수축 억제능을 관찰하였고 아직 보고가 빈약한 수축성 조절 기전에 대해 조직 선택적 조절 가설을 세워서 조사하였다. 내피가 손상된 혈관을 수조내에 현수시켰고 혈관에 의한 기계적 신호가 등장력 변환기에서 전기적 신호로 변환되 어 생리측정기에 표시되었다. 내피가 손상된 혈관에서 phenylephrine은 조직에 비선택적으로 지속적인 수축을 유지하였고 phenylephrine과 병용된 isoprenaline은 등척성 수축 실험에서 흉부, 복부 대동맥 등 조직에 비선 택적으로 평활근에 대한 직접 작용으로 수축성을 일시적으로 감소시켰고 phenylephrine과 병용된 prazosin은 조직에 비선택적으로 평활근에 대한 직접 작용으로 수축성을 지속적으로 감소시켰다. 따라서 일부 조직에서 adrenergic beta receptor 밀도가 감소되거나 수용체 결합력이 감소되거나 수용체 결합 후 신호전달이 감소되 거나 약물의 분포가 감소되어 isoprenaline이 phenylephrine의 지속적 작용에 대해 일시적으로 억제하고 prazosin이 지속적으로 억제하는 것으로 생각된다. In the study, we endeavored to investigate the effect of phenylephrine, isoprenaline and prazosin on the tissue-specific vascular contractility and to determine the mechanism involved. There were few reports addressing the question whether thin or thick filament modulation is included in phenylephrine, isoprenaline and prazosin-induced regulation. We hypothesized that isoprenaline and prazosin play a role in tissue-dependent regulation of vascular contractility. Denuded arterial muscles of Sprague-Dawley male rats were suspended in organ baths and isometric tensions were transduced and recorded using isometric transducers and an automatic data acquisition system. Interestingly, sustained continuous contraction of thoracic and abdominal aorta. Furthermore, isoprenaline and prazosin together with phenylephrine inhibited transiently and persistently vasoconstriction of thoracic and abdominal aorta suggesting that additional mechanisms (e.g. decreased receptor density, chemical interaction, postreceptor signaling or distribution of agonists) might be included in the modulation of vascular contractility.

엉겅퀴 유래 Silymarin의 단독 및 알코올 병용 시 혈압 조절 효과

제현동,민영실 대한산업경영학회 2022 산업융합연구 Vol.20 No.7

In the study, we endeavored to assess the convergence effect of Silybum marianum-derived silymarin and epidemiologically-correlated alcohol intake on vascular contractility and to determine the mechanism involved. There were few reports addressing the question whether thin or thick filament modulation is included in ethanol and silymarin-induced regulation. We hypothesized that ethanol at a low concentration and silymarin play a role in agonist-dependent regulation of vascular contractility. Denuded arterial muscles of Sprague-Dawley male rats were suspended in organ baths and isometric tensions were transduced and recorded using isometric transducers and an automatic data acquisition system. Interestingly, both silymarin and ethanol didn’t encourage silymarin alone-induced inhibition in agonists-induced contraction suggesting that endothelial nitric oxide synthesis might be involved in ethanol or silymarin-induced modulation of vascular contractility and additional pathways besides endothelial nitric oxide synthesis such as ROCK inactivation might be involved in the silymarin-induced modulation of vascular contractility. 역학 조사에서 알코올 섭취와 고혈압 증가 사이에 인과관계가 있어서 이번 연구에서 엉겅퀴 유래 silymarin의 단독 및 알코올 병용 투여에서 혈관수축 억제능을 관찰하였고 아직 불분명한 수축성 조절 기전에 대해 효능제 선택적 조절 가설을 수립하여 조사하였다. 내피가 손상된 혈관이 수조 내 현수되었고 혈관에 의한 기계적 신호가 등장력 변환기에서 전기적 신호로 변환되어 생리측정기에 표시되었다. 저농도의 ethanol과 silymarin은 혈관 내피에서 산화질소 생성 작용 외에 평활근에 대한 직접 작용으로 동맥의 수축성을 감소시킬 것으로 추측되었는데 인위적으로 내피가 손상된 동맥에서 ethanol과 병용된 silymarin이 silymarin 단독에 비해 굵은 미세섬유성 조절성 수축약 (fluoride, thromboxane mimetic)에 의한 혈관 수축 억제에 차이가 없었고 silymarin 단독에 비해 가는 미세섬유성 조절성 phorbol ester에 의한 혈관 수축 억제에 차이가 없었다. 따라서 silymarin 단독은 내피 의존성 산화질소 생성과 내피에 비의존적으로 평활근에서 주로 ROCK 활성 감소에 참여하여 결과적으로 평활근에서 악틴-미오신 상호작용을 억제하여 혈관을 이완시키고 ethanol은 내피 의존성 산화질소 생성 외에 평활근에 대한 작용이 없는 것으로 생각된다.

혈관수축에 대한 감초산 Glycyrrhizic acid의 효과

제현동,민영실 대한산업경영학회 2023 산업융합연구 Vol.21 No.10

This study aims to elucidate the effect of glycyrrhizic acid on smooth muscle contraction and to determine the detailed mechanism incorporated. We hypothesized that glycyrrhizic acid played a role in the agonist-sensitive management of smooth muscle contraction. Stripped smooth muscles of Sprague-Dawley rats were prepared in organ baths and isometric tensions were converted, stored and analyzed by using isometric transducers, a physiograph and one way ANOVA. Interestingly, glycyrrhizic acid attenuated the thick filament regulating agonist (fluoride or thromboxane mimetic)-sensitive contraction (p=0.113, 0.008, 0.004 (Student's t-test), p=0.113, 0.008, 0.004 (One way ANOVA) at 0.01, 0.03, 0.1 mM fluoride, and p=0.156, 0.004, 0.003 (Student's t-test), p=0.156, 0.004, 0.003 (One way ANOVA) at 0.01, 0.03, 0.1 mM thromboxane mimetic) and did not attenuate the thin filament regulating agonist (phorbol ester)-induced contraction (p=0.392, 0.086, 0.065 (Student's t-test), p=0.392, 0.086, 0.065 (One way ANOVA) at 0.01, 0.03, 0.1 mM phorbol ester). It is suggesting that endothelial EDRF (NO) synthesis and accessory pathways besides endothelial EDRF (NO) synthesis such as ROCK restriction might be incorporated in the glycyrrhizic acid-induced modulation of smooth muscle contraction inhibiting acto-myosin interaction.

위장관 수축성에 대한 Histamine과 Atropine의 융합성 조절 효과

제현동,민영실,Je, Hyun Dong,Min, Young Sil 중소기업융합학회 2021 융합정보논문지 Vol.11 No.10

본 연구에서 진단, 과민반응 검사에 사용되며 의약품에 불순물로 포함되어 규제되는 histamine의 단독 및 병용 투여에서 조직/근육 선택적 장관 수축성 조절에 대한 실험을 하고자 하였다. Histamine은 muscarinic receptor 작용 외에 histamine H1 receptor에 대한 직접 작용으로 수축성을 증가시켰고 회장/결장 윤주근에서 수축성을 거의 증가시키지 않았다. 또한 M3 수용체에 선택적으로 작용하는 atropine의 병용 효과를 관찰하였는데 histamine과 병용된 atropine은 장관 중 회장의 종주근의 수축성을 거의 감소시키지 않았고 회장/결장의 윤주근의 수축성을 감소시키지 않았다. 따라서 적어도 회장의 종주근에서 histamine은 M3 수용체를 거의 경유하지 않고 작용하고 atropine은 항무스카린 효과 외에 histamine에 의한 조직 선택적 위장관 운동성을 거의 조절하지 않는 것으로 생각된다. 그리고 이러한 단순한 검색은 의약품 불순물 검사로 선호될 것으로 생각된다. The aim of the study was to observe the influence and related mechanism of histamine and its analogues used for hypersensitivity tests and used as an indicator of impurities in drugs on the tissue-specific intestinal contraction. Intestinal contraction includes the activation of thick or thin filament regulation. However, there are few reports addressing the question whether this regulation is involved in histamine-induced regulation. We hypothesized that histamine plays a role in tissue-dependent regulation of intestinal contractility. Denuded ileal/colonic longitudinal and circular muscles of male rats were used and isometric contractions were recorded using a data acquisition system. Interestingly, histamine alone didn't increase the contraction of the circular muscle but increased the contraction of the longitudinal muscle. Histamine together with atropine (M3 receptor antagonist) didn't inhibit the contraction of the longitudinal and circular muscle. Therefore, histamine alone and together with atropine increases the ileal longitudinal muscle contraction suggesting that additional mechanisms (decreased receptor density, postreceptor signaling or distribution of agonists) might be involved in the regulation of ileal muscle contractility. In conclusion, histamine and/or atropine has some effect on the regulation of the longitudinal contractility regardless of M3 receptor and the simpler test would be preferred as the drug impurity test compared to more complicated tests.