관상동맥이완과 혈소판응집에 대한 GS283 과 GS386 의 약리작용기전에 관한 연구

장기철(Ki Churl Chang),이회영(Hoi Young Lee),이균우(Goun Woo Lee),구의본(Eui Bon Koo),강영진(Young Jin Kang),이영수(Young Soo Lee) 한국응용약물학회 1997 Biomolecules & Therapeutics(구 응용약물학회지) Vol.5 No.3

Trimetoquinol (TMQ) and its analogs are known to have thromboxane A₂ antagonistic action. We also reported that GS389, chemically similar to TMQ, has competitive antagonistic action in rat aorta and human platelets. In the present study, we investigated the pharmacological characteristics of GS283 and GS386, analogs of GS389, using vascular smooth muscle, human platelets and rat brain homogenates. In isolated pig coronary artery (PCA), both of GS283 and GS386 relaxed U46619-contracted rings in concentration dependent manner. Pretreatment with several concentrations of GS283 and GS386 shifted the dose-response curves to the right, and reduced of maximum contration dose-dependently. Furthermore, GS283 and GS386 strongly inhibited Ca^(2+)-induced contraction in the PCA. In human platelets, U46619- and A23187-induced platelet aggregation was inhibited by GS283 and GS386, concentration-dependently. Anti-platelet aggregation was related to the compound`s ability to inhibit ATP release at each stimulation. In rat brain homogenates, receptor-binding assay resulted that both GS283 and GS386 have a relative affinity to α-adrenergic receptor. Taken together, we concluded that the mechamism of action of GS283 and GS386 is not related with in TXA₂ receptor but concerned with calcium antagonistic action and α-blocking action.

Calcium Channel Blocking and Phosphodiesterase Inhibitory Action of GS386, a Dihydroisoquinoline Derivative, in Isolated Rat Trachea

장기철,이회영,강영진,구의본,Chang, Ki-Churl,Lee, Hoi-Young,Kang, Young-Jin,Koo, Eui-Bon The Korean Society of Pharmacology 1996 대한약리학잡지 Vol.32 No.3

최근 본 연구실에서는 GS 386인 1-(4'-methoxybenzyl)-6,7-dimethoxy-3,4-dihydroisoquinoline이 적출된 토끼의 심방세포에서 $Ca^{++}$ 채널의 운동성 변화없이 $Ca^{++}$ 채널이 열릴 가능성을 줄임으로써 $Ca^{++}$ 전류의 증폭을 억제한다고 보고하였다. 이번 연구에서는 적출된 쥐의 기관지를 사용하여 GS 386의 작용기전에 대해 연구하였다. GS386은 carbachol $(0.3{\mu}M)$과 높은 농도의 $K^+$ (65.4mM)에 의해 수축된 쥐의 기관지를 용량-의존적으로 이완시켰으며 이때 $IC_{50}$는 5.24와 $5.67\;{\mu}M$이었다. verapamil은 carbachol에 의한 수축시 보다 높은 농도의 $K^+$에 의해 수축된 조직에 더욱 효과적으로 억제하였다. $Ca^{++}$이 없는 상태에서 $Ca^{++}$에 의한 수축은 GS386에 의해 억제되었다. 더욱이 높은 농도의 GS386$(100\;{\mu}M)$은 verapamil과는 다르게 carbachol뿐만 아니라 caffeine에 의한 위상성 수축을 억제 시키므로 GS386은 세포질내로 들어가 sarcoplasmic retuculum과 같은 근육 내부에 2차적인 영향을 나타내었다. 더군다나GS386은 verapamil에 의해 영향을 받지않는 (verapamil-insensitive component)이완을 보였고 쥐 기관지의 평활근에서 cAMP의 양을 증가 시켰다. 이러한 결과는 GS386의 작용기전이 $Ca^{++}$ 길항적인 작용 뿐만 아니라 posphodiesterase억제작용에 기인한다는 사실을 제시한다. Recently we reported that GS 386, 1-(4'-methoxybenzyl)-6,7-dimethoxy-3,4-dihydroisoquinoline, inhibited amplitude of the $Ca^{2+}$ current by reducing the probability of $Ca^{2+}$ channel opening without changing channel kinetics in isolated rabbit atrial myocyte. In the present study, further investigation of the mechanism of action of GS 386 was performed using isolated rat trachealis. GS 386 concentration-dependently relaxed rat trachealis contracted by carbachol $(0.3{\mu}M)$ and high $K^+$(65.4 mM) with $IC_{50}$ 5.24 and 5.67 ${\mu}M$, respectively. Verapamil inhibited more effectively the high $K^+-contracted$ tissues than those with carbachol in the rat trachealis muscle. In $Ca^{2+}-free$ media, $Ca^{2+}-induced$ contraction was inhibited by GS 386. Furthermore, high concentration of GS 386 $(100{\mu}M)$ but not verapamil, attenuated a phasic contraction induced not only by carbachol but also caffeine, indicating that GS386 can enter into the cytoplasm where it may exert secondary actions on internal sites of the muscle, such as sarcoplasmic reticulum. Moreover, GS 386 showed verapamil-resistant component of relaxation and increased cAMP levels in rat trachal smooth muscle. These results suggest that the mechanism of action of GS 386 attributes to not only $Ca^{2+}$ antagonistic action but also weak phosphodiesterase inhibitory action.

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.

돼지 간장 조직내 Protein Methylase Ⅱ 저해제의 정제 및 특성

권명희,정기경,이회영,이향우,홍성렬 ( Myung Hee Kwon,Ki Kyung Jung Hoi,Young Lee,Hyang Woo Lee,Sungyoul Hong ) 생화학분자생물학회 1994 BMB Reports Vol.27 No.6

An inhibitor for protein methylase II (EC 2.1.1.24) was solubilized from porcine liver microsomal fraction by heat treatment, and purified by ultrafiltration, Sephadex G-25 chromatogrnphy, and HPLC using μ-Bondapak C_(18) column. The purified inhibitor was near homogeniety as judged by HPLC. The molecular weight of the inhibitor was estimated to be 1,676 Da by analysis of amino acid composition. And it was revealed that the inhibitor molecule is rich in alanine and glycine. The activity of the inhibitor was not affected by heat treatment up to 100℃ as well as hydrolytic enzymes. The K; value for the protein methylase II which has been purified from porcine spleen was measured to be 1.3 × 10^(-7) M. Inhibition studies showed that the inhibitor was noncompetitive with respect to S-adenosyl-L-methionine (SAM) and activities of several SAM-dependent methylases were also inhibited by the purified inhibitor.

Lipopolysaccharide로 활성화시킨 흰쥐 혈관의 iNOS 발현에 대한 Higenamine의 효과

강영진,이균우,구의본,이회영,장기철,Kang Young-Jin,Lee Goun-Woo,Ku Eui-Bon,Lee Hoi-Young,Chang Ki-Churl 대한약리학회 1997 The Korean Journal of Physiology & Pharmacology Vol.1 No.3

Higenamine was widely used as traditional remedy for the treatment of rhumatoid arthritis. Nitric oxide(NO) may be a critical mediator in this inflammatory disease. Synovial tissue from humans with inflammatory arthritis expresses NOS2(iNOS) mRNA and protein, and generates NO in vitro. We therefore, investigated the effect of higenamine on the induction of nitric oxide synthase(NOS) promoted by lipopolysaccharide(LPS). Prophylactic application of higenamine selectively prevented LPS-primed initiation of L-arginine-induced relaxation and restored rhenylephrine(PE)-induced contraction in rat aorta. LPS-stimulated nitrite production in the incubation medium was reduced by higenamine. Furthermore, RT-PCR and Northern analysis indicated that higenamine reduced iNOS expression primed by LPS in rat aorta. These results suggest that higenamine prevents LPS-promoted induction of NOS in vascular smooth 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.

낫토균으로 발효한 발효울금의 투여가 마우스의 간 기능 및 혈중 지질 함량에 미치는 영향

강재구(Jae Ku Kang),강효진(Hyo Jin Kang),서지혜(Ji Hye Seo),김선옥(Sun Ok Kim),최정효(Jung Hyo Choi),조도연(Do Yeun Cho),박창교(Chang Gyo Park),이회영(Hoi Young Lee) 한국식품영양과학회 2009 한국식품영양과학회지 Vol.38 No.4

본 연구는 낫토균으로 울금을 발효함으로써 울금의 기호성 향상 및 낫토키나아제의 작용으로 울금의 체내이용율을 증가시켜 생리활성 기능이 증가하는지 알아보고자 연구하였다. 울금을 낫토균으로 발효시킨 결과, 칼슘과 아연의 경우 울금에 비하여 발효울금에서 각각 약 6배와 44배 증가를 보였고, 셀레늄의 경우에는 발효울금에서만 새롭게 검출되었다. 항산화 활성 비교실험 결과, 울금과 발효울금의 항산화 활성은 각각의 실험농도에서 비슷한 효과를 나타내었으나 발효울금이 울금보다 항산화 활성이 약간 높았다. 발효울금의 간 기능 개선효과에 미치는 영향을 사염화탄소로 간손상을 일으킨 마우스에서 실험한 결과, AST 및 ALT 효소활성의 유의적인 감소를 보였고(p<0.05) 특히 사염화탄소에 의해 야기된 간조직의 염증, 괴사 및 지방의 축적이 확연히 적어지는 등 정상군에 가깝게 간 손상이 회복되어 발효울금은 간 기능성 개선에서 울금보다 효과적이었다. 낫토균으로 발효한 발효울금의 혈중 지질 개선효과를 평가하기 위해 고지방(20% 지방) 식이로 유도된 마우스에서 발효울금 및 울금의 경구투여에 의한 혈중 지질 함량 변화를 평가한 결과, 고지방 식이에 의해 증가된 총 콜레스테롤과 LDL 농도를 유의적으로 감소시켰으며 HDL 콜레스테롤 농도는 유의하게 증가시켰다(p<0.05). 본 연구에서 울금 및 발효울금 모두 혈중지질을 효과적으로 개선하였으나, 발효울금의 경우 저농도 투여에 의해서도 혈중 지질 개선효과가 나타나 울금에 비해 상대적인 우수성을 나타내었다. 또한 낫토균에 의한 울금의 발효가 울금 특유의 이취에 의한 쓴맛 및 신맛을 유의하게 개선하는 것으로 평가되어 적절한 제품화 과정을 거친다면 기능뿐만 아니라 소비자의 기호를 향상시킬 수 있는 건강 기능성 식품개발에 기여할 것으로 생각된다. The effects of turmeric and fermented turmeric by Bacillus natto on antioxidant activities, liver function recovery of acute hepatotoxicity mice, and serum lipid parameters in high fat diet fed mice were investigated. In the results of antioxidant activity by DPPH method, fermented turmeric had higher antioxidative activity than turmeric. Acute hepatotoxicity was induced by 0.5 ㎖ of carbon tetrachloride (CCl₄) per kg of mice. Unlike turmeric, fermented turmeric significantly reduced the levels of aspartate aminotransferase (AST) and alanine aminotransferase (ALT) after 5 days compared to the controls with 0.5% methyl cellulose (p<0.05). In addition, higher recovery of liver damage by CCl₄ was observed in mice with fermented turmeric than with turmeric. High fat (20%) diet fed mice were divided into 4 groups to investigate the effects of turmeric and fermented turmeric on serum lipid parameters: C (vehicle), TuL (low dose (80 ㎎/㎏) with turmeric), TuH (high dose (160 ㎎/㎏) with turmeric), FTuL (low dose with fermented turmeric), and FTuH (high dose with fermented turmeric). The levels of LDL-cholesterol and HDL-cholesterol were significantly reduced and increased in FTuL, FTuH and TuH groups compared to the C group, respectively. However, there was no significant change in triglyceride levels by either turmeric or fermented turmeric compared to those by control. Collectively, these results strongly suggest that fermented turmeric by Bacillus natto could be used as a functional food for enhancement of health with better consumer acceptance.

膵臟 組織내 Protein Methylase I 酵素의 精製

李香雨,李晦榮 成均館大學校 科學技術硏究所 1985 論文集 Vol.36 No.1

An enzyme which methylates the guanido group of arginine residues of protein using S-adenosyl-L-methionine as methyl donor has been designated as protein methylase I. Among the enzyme products of this reaction which thus far have been identified are N^G-mono, N^G,N^c-di-, and N^G, N'^G- dimethylarginine. This enzyme is widely distributed in various organs of rat, being especially rich in pancreas of hog. The level of protein methylase I activity was found to closely parallel the rate of cell proliferation. Elevated activity was observed in fetal tissues, continuously dividing HeLa S-3 cell culture, regenerating adult rat liver, and in fast growing Novikoff hepatoma. Protein merhylase I has been purified from calf brain, rat and calf thymus, Krebs II ascites cells and recently a form of protein merhylase I has also been purified from wheat germ, which is the first report of the enzyme being present in the plant kingdom. In the present paper, protein methylase I has been partially purified from hog pancreatic tissue. 1) Protein methylase I has been partially purified about 14-fold from hog pancreatic tissue with 25% yield. 2) Mainly the enzyme was found in cytosolic fraction of pancreatic cells. 3) Main purification steps were homogenation of the tissue, S_3 fraction of cells, 35% ammonium sulfate ppt., and DIEM-cellulose chromatography.