• Journal of Ginseng Research
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• 제35권2호
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• pp.176-190
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• 2011

There seems to be some controversy about the effect of total ginseng saponin (TGS) on the secretion of catecholamines (CA) from the adrenal gland. Therefore, the present study aimed to determine whether TGS can affect the CA release in the perfused model of the adrenal medulla isolated from spontaneously hypertensive rats (SHRs). TGS (15-150 ${\mu}g/mL$), perfused into an adrenal vein for 90 min, inhibited the CA secretory responses evoked by acetylcholine (ACh, 5.32 mM) and high $K^+$ (56 mM, a direct membrane depolarizer) in a dose- and time-dependent fashion. TGS (50 ${\mu}g/mL$) also time-dependently inhibited the CA secretion evoked by 1.1-dimethyl-4 -phenyl piperazinium iodide (DMPP; 100 ${\mu}M$, a selective neuronal nicotinic receptor agonist) and McN-A-343 (100 ${\mu}M$, a selective muscarinic M1 receptor agonist). TGS itself did not affect basal CA secretion (data not shown). Also, in the presence of TGS (50 ${\mu}g/mL$), the secretory responses of CA evoked by veratridine (a selective $Na^+$ channel activator (50 ${\mu}M$), Bay-K-8644 (an L-type dihydropyridine $Ca^{2+}$ channel activator, 10 ${\mu}M$), and cyclopiazonic acid (a cytoplasmic $Ca^{2+}$-ATPase inhibitor, 10 ${\mu}M$) were significantly reduced, respectively. Interestingly, in the simultaneous presence of TGS (50 ${\mu}g/mL$) and N${\omega}$-nitro-L-arginine methyl ester hydrochloride [an inhibitor of nitric oxide (NO) synthase, 30 ${\mu}M$], the inhibitory responses of TGS on the CA secretion evoked by ACh, high $K^+$, DMPP, McN-A-343, Bay-K-8644, cyclopiazonic acid, and veratridine were considerably recovered to the extent of the corresponding control secretion compared with the inhibitory effect of TGS-treatment alone. Practically, the level of NO released from adrenal medulla after the treatment of TGS (150 ${\mu}g/mL$) was greatly elevated compared to the corresponding basal released level. Taken together, these results demonstrate that TGS inhibits the CA secretory responses evoked by stimulation of cholinergic (both muscarinic and nicotinic) receptors as well as by direct membrane-depolarization from the isolated perfused adrenal medulla of the SHRs. It seems that this inhibitory effect of TGS is mediated by inhibiting both the influx of $Ca^{2+}$ and Na+ into the adrenomedullary chromaffin cells and also by suppressing the release of $Ca^{2+}$ from the cytoplasmic calcium store, at least partly through the increased NO production due to the activation of nitric oxide synthase, which is relevant to neuronal nicotinic receptor blockade, without the enhancement effect on the CA release. Based on these effects, it is also thought that there are some species differences in the adrenomedullary CA secretion between the rabbit and SHR.

• 대한약리학회지
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• 제24권2호
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• pp.203-216
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• 1988

돼지, 가표 및 가토의 뇌동맥에서 기저동맥 (basilar artery, BA)과 circle of willis동맥 (WC)의 동맥환을 만들어 혈관수축제와 calcium 길항제의 효과 그리고 내피세포의 역할은 검토하였다. 돼지 BA와 WC에 서 norepinephrine (NE), phenylephrine (PE)및 epinephrine (EP)은 propranolol $10^{-6}M$ 전처리하에서 그리고 KCI, histamine, 5-hydroxytryptamine (5-HT) 및 angiotensin은 모두 용량의존성 수축반응을 일으켰다. 그 최대수축력은 angiotensin에서 가장 적었고, 5-HT에서 가장 강력하였다. KCI 35 mM수축반응은 calcium 길항제로 용량의존성으로 억제되었고, 그 효력은 nifedipine > > diltiazem > flunarizine > oxybutynin > isosorbide dinitrate (ISDN) > glycerl trinitrate의 순서였다. 5-HT $10^{-5}M$의 수축반응은 nifedipine으로는 용량의존성으로 억제 되었으나 diltiazem과 ISDN으로는 경미한 억제만을 일으켰다. 내피세포동맥환에서 KCI 35 mM의 수축반응은 acetylcholine (ACh)으로 거의 영향받지 않았으나 $PGF_{2{\alpha}}\;10^{-5}M$의 수축반응은 ACh과 adenosine으로 용량의존성으로 억제되었고, 이 내피세포 의존성억제는 nifedipine $10^{-6}M$로는 영향받지 않으나 methylene blue $50\;{\mu}M$로는 현저히 억제되었다. 네피제거동맥환에의 $PGF_{2{\alpha}}$의 수축반응은 ACh의 영향이 없거나 더욱 수축되었다. 내피세포를 제거하지 않은 가토흉부대동맥편을 bath에 함께 넣어주면 ACh의 용량의존성 이완반응이 나타났고 이 이완반응도methylene blue로 억제되었다. 가묘 BA와 WC에서 5-HT와 NE는 용량의존성 수축반응을 일으켰고 ACh도 내피세포 존재유무와 관계없이 강력한 용량의존성 수축반응을 일으켰으며 그 최대수축력은 ACh에서 가장 강력하였다. 내피세포동맥환에서 5-HT $10^{-5}M$의 수축반응은 ACh에 의하여 이완반응없이 더욱 수측되었다. 가토 BA에서 5-HT와 NE는 응량의존성 수축반응을 일으졌고5-HT의 수축반응이 더욱 현저하였다. 내피세포동맥환에서 5-HT $10^{-5}M$의 수축반응은 ACh $10^{-5}M$로 현저하게 이완되있고 이 내피세포의존성 이완반응은 atropine $10^{-7}M$로는 억제되었으나 diltiazem $10^{-6}M$로는 거의 억제되지 않았다. 이상의 실험성적은 돼지와 가토의 뇌동맥에서 ACh은 내피세포-의존성 이완반응을 일으키며그 이완반응은 muscarinic receptor를 통해 나타났고, 가묘뇌동맥에서 ACh은 혈관수축제의 역할을 갖고 있음을 시사하고 있다.

• 대한약리학회지
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• 제24권1호
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• pp.93-101
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• 1988

돼지 우관상동백을 적출하여 항경련제인 oxybutynin의 약리작용을 조사하였다. 1. Acetylcholine (ACh)과 KCl은 관상동맥을 수축시켰고 이 수축효과는 용량의존적이었다. ACh의 수축효과는 내피손상표본 $(EC_{50}=0.52\;{\mu}M)$에서 내피표본 $(EC_{50}=0.52\;{\mu}M)$보다 약 2배 강화되있으나 KCl의 수축효과는 양군간에 차이가 없었다. 2. $ACh(1.0\;{\mu}M)$의 수축효과는 oxybutynin과 atropine에 의 하여 용량의존적으로 억제되었고 두 약물의 $(IC_{50}$는 각각 11.0 nM과 0.47 nM로 atropine이 약 23배나 더 예민하였다. 그러나 KCl (35 mM)의 수축효과는 atropine으로는 전혀 영향받지 않았고 oxybutynin으로는 용량의존적으로 억제 되었으며 $(IC_{50}=49.7\;{\mu}M)$이였다. 3. ACh의 용량반응곡선은 oxybutynin $(IC_{50}=11\;nM)$ 및 atropine $(IC_{50}=0.47\;nM)$ 전처 리 하에서 우측으로 평 행 이 동되 있고, KCI의 용량반응곡선은 oxybutynin $(IC_{50}=49.7\;nM)$ 전처리하에서 우측으로 비상경적 이동을 일으켰다. 4. Oxybutynin은 Bay K 8644 $(0.1\;{\mu}M)$의 수축효과를 용량의존적 으로 억제하였고 $(IC_{50}=63.0\;{\mu}M)$이었으며 , histamine $(35\;{\mu}M)$의 수축효과는 oxybutynin의 최대량 $(500\;{\mu}M)$으로 부분억제 (최대 50%)만을 일으켰다. 이상의 성적으로 적출돼지 관상동백에서 내피세포는 ACh에 의한 수축반응에 억제적 영향을 미치며, oxybutynin은 강력한 muscarine receptor 차단작용과 calcium influx 억제작용에 의하여 혈관근 이완을 일으킨다고 추론하였다.

• The Korean Journal of Physiology and Pharmacology
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• 제8권5호
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• pp.265-272
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• 2004

The present study was attempted to investigate the effect of cilnidipine (FRC-8635), which is a newly synthesised novel dihydropyridine (DHP) type of organic $Ca^{2+}$ channel blockers, on secretion of catecholamines (CA) evoked by acetylcholine (ACh), high $K^+$, DMPP and McN-A-343 from the isolated perfused rat adrenal gland. Cilnidipine $(1{\sim}10{\mu}M)$ perfused into an adrenal vein for 60 min produced relatively dose- and time-dependent inhibition in CA secretory responses evoked by ACh $(5.32{\times}10^{-3}M),\;DMPP\;(10^{-4}M\;for\;2\;min)$ and McN-A-343 $(10^{-4}M\;for\;2\;min)$. However, lower dose of cilnidipine did not affect CA secretion by high $K^+\;(5.6{\times}10^{-2}\;M)$, higher dose of it reduced greatly CA secretion of high $K^{+}$. Cilnidipine itself did fail to affect basal catecholamine output. In the presence of cilnidipine $(10{\mu}M)$, the CA secretory responses evoked by Bay-K-8644 $(10{\mu}M)$, an activator of L-type $Ca^{2+}$ channels and cyclopiazonic acid $(10{\mu}M)$, an inhibitor of cytoplasmic $Ca^{2+}$-ATPase were also inhibited. Moreover, ${\omega}-conotoxin\;GVIA\;(1{\mu}M)$, a selective blocker of the N-type $Ca^{2+}$ channels, given into the adrenal gland for 60 min, also inhibited time-dependently CA secretory responses evoked by Ach, high $K^+$, DMPP, McN-A-343, Bay-K-8644 and cyclopiazonic acid. Taken together, these results demostrate that cilnidipine inhibits CA secretion evoked by stimulation of cholinergic (both nicotinic and muscarinic) receptors from the isolated perfused rat adrenal gland without affecting the basal release. However, at lower dose, cilnidipine did not affect CA release by membrane depolarization while at larger dose inhibited that. It seems likely that this inhibitory effect of cilnidipine is exerted by blocking both L- and N-type voltage-dependent $Ca^{2+}$ channels (VDCCs) on the rat adrenomedullary chromaffin cells, which is relevant to inhibition of both the $Ca^{2+}$ influx into the adrenal chromaffin cells and intracellular $Ca^{2+}$ release from the cytoplasmic store. It is thought that N-type VDCCs may play an important role in regulation of CA release from the rat adrenal medulla.

• The Korean Journal of Physiology and Pharmacology
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• 제20권6호
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• pp.629-639
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• 2016

The present study was designed to investigate the characteristics of gintonin, one of components isolated from Korean Ginseng on secretion of catecholamines (CA) from the isolated perfused model of rat adrenal gland and to clarify its mechanism of action. Gintonin (1 to $30{\mu}g/ml$), perfused into an adrenal vein, markedly increased the CA secretion from the perfused rat adrenal medulla in a dose-dependent fashion. The gintonin-evoked CA secretion was greatly inhibited in the presence of chlorisondamine ($1{\mu}M$, an autonomic ganglionic bloker), pirenzepine ($2{\mu}M$, a muscarinic $M_1$ receptor antagonist), Ki14625 ($10{\mu}M$, an $LPA_{1/3}$ receptor antagonist), amiloride (1 mM, an inhibitor of $Na^+/Ca^{2+}$ exchanger), a nicardipine ($1{\mu}M$, a voltage-dependent $Ca^{2+}$ channel blocker), TMB-8 ($1{\mu}M$, an intracellular $Ca^{2+}$ antagonist), and perfusion of $Ca^{2+}$-free Krebs solution with 5mM EGTA (a $Ca^{2+}$chelater), while was not affected by sodium nitroprusside ($100{\mu}M$, a nitrosovasodialtor). Interestingly, LPA ($0.3{\sim}3{\mu}M$, an LPA receptor agonist) also dose-dependently enhanced the CA secretion from the adrenal medulla, but this facilitatory effect of LPA was greatly inhibited in the presence of Ki 14625 ($10{\mu}M$). Moreover, acetylcholine (AC)-evoked CA secretion was greatly potentiated during the perfusion of gintonin ($3{\mu}g/ml$). Taken together, these results demonstrate the first evidence that gintonin increases the CA secretion from the perfused rat adrenal medulla in a dose-dependent fashion. This facilitatory effect of gintonin seems to be associated with activation of LPA- and cholinergic-receptors, which are relevant to the cytoplasmic $Ca^{2+}$ increase by stimulation of the $Ca^{2+}$ influx as well as by the inhibition of $Ca^{2+}$ uptake into the cytoplasmic $Ca^{2+}$ stores, without the increased nitric oxide (NO). Based on these results, it is thought that gintonin, one of ginseng components, can elevate the CA secretion from adrenal medulla by regulating the $Ca^{2+}$ mobilization for exocytosis, suggesting facilitation of cardiovascular system. Also, these findings show that gintonin might be at least one of ginseng-induced hypertensive components.

• Journal of Pharmaceutical Investigation
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• 제32권1호
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• pp.47-54
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• 2002

Cimetropium bromide, a quaternary ammonium compound which is chemically related to scopolamine, exhibits its antispasmodic activity by competing with acetylcholine for the muscarinic receptors of the smooth muscle of gastrointestinal tract. The drug has been used for the treatment of various disorders involving spasms of the musculature of the gastrointestinal, biliary and genitourinary tracts. The purpose of the present study was to evaluate the bioequivalence of two cimetropium bromide tablets, $Algiron^{TM}$ (Boehringer Ingelheim Korea Ltd.) and $Alpit^{TM}$ (Hana Pharmaceutical Co., Ltd.), according to the prior and revised guidelines of Korea Food and Drug Administration (KFDA). The cimetropium bromide release from the two cimetropium bromide tablets in vitro was tested using KP VII Apparatus II method with various different kinds of dissolution media (pH 1.2, 4.0, 6.8 buffer solution and water). Twenty normal male volunteers, $25.25{\pm}2.10$ years in age and $65.76{\pm}6.39$ kg in body weight, were divided into two groups and a randomized $2{\times}2$ cross-over study was employed. After three tablets containing 50 mg of cimetropium bromide per tablet were orally administered, blood was taken at predetermined time intervals and the concentrations of cimetropium bromide in serum were determined using HPLC method with UV detector. The dissolution profiles of two cimetropium bromide tablets were very similar at all dissolution media. Besides, the pharmacokinetic parameters such as $AUC_t,\;C_{max}\;and\;T_{max}$ were calculated and ANOVA test was utilized for the statistical analysis of the parameters using non-transformed and logarithmically transformed $AUC_t\;and\;C_{max}$. The results showed that the differences in $AUC_t,\;C_{max}\;and\;T_{max}$ between two tablets based on the $Algiron^{TM}$ were 2.19%, -5.97% and 3.49%, respectively. Minimum detectable differences $({\Delta})\;at \;{\alpha}=0.05\;and\;1-{\beta}=0.8$ were less than 20% (e.g., 13.71 %, 19.05% and 15.11% for $AUC_t,\;C_{max}\;and\;T_{max}$, respectively). The powers $(1-{\beta})\;at\;{\alpha}=0.05,\;{\Delta}=0.2\;for\;AUC_t$, $C_{max}\;and\;T_{max}$ were 97.79%, 83.22% and 95.60%, respectively. The 90% confidence intervals were within ${\pm}20%$ (e.g., $-5.84{\sim}10.21,\;-17.11{\sim}5.18\;and\;-5.35{\sim}12.33\;for\;AUC_t,\;C_{max}\;and\;T_{max}$, respectively). There were no sequence effect between two tablets in logarithmically transformed $AUC_t\;and\;C_{max}$. The 90% confidence intervals using logarithmically transformed data were within the acceptance range of log(0.8) to log(1.25) (e.g., $0.94{\sim}1.10\;and\;0.85{\sim}1.05\;for\;AUC_t\;and\;C_{max}$, respectively). Two parameters met the criteria of prior and revised KFDA guideline for bioequivalence, indicating that $Alpit^{TM}$ tablet is bioequivalent to $Algiron^{TM}$ tablet.

• IMMUNE NETWORK
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• 제21권5호
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• pp.34.1-34.16
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• 2021

Sjögren's syndrome (SS) is an autoimmune disease characterized by dryness of the mouth and eyes. The glandular dysfunction in SS involves not only T cell-mediated destruction of the glands but also autoantibodies against the type 3 muscarinic acetylcholine receptor or aquaporin 5 (AQP5) that interfere with the secretion process. Studies on the breakage of tolerance and induction of autoantibodies to these autoantigens could benefit SS patients. To break tolerance, we utilized a PmE-L peptide derived from the AQP5-homologous aquaporin of Prevotella melaninogenica (PmAqp) that contained both a B cell "E" epitope and a T cell epitope. Repeated subcutaneous immunization of C57BL/6 mice with the PmE-L peptide efficiently induced the production of Abs against the "E" epitope of mouse/human AQP5 (AQP5E), and we aimed to characterize the antigen specificity, the sequences of AQP5E-specific B cell receptors, and salivary gland phenotypes of these mice. Sera containing anti-AQP5E IgG not only stained mouse Aqp5 expressed in the submandibular glands but also detected PmApq and PmE-L by immunoblotting, suggesting molecular mimicry. Characterization of the AQP5E-specific autoantibodies selected from the screening of phage display Ab libraries and mapping of the B cell receptor repertoires revealed that the AQP5E-specific B cells acquired the ability to bind to the Ag through cumulative somatic hypermutation. Importantly, animals with anti-AQP5E Abs had decreased salivary flow rates without immune cell infiltration into the salivary glands. This model will be useful for investigating the role of anti-AQP5 autoantibodies in glandular dysfunction in SS and testing new therapeutics targeting autoantibody production.

• 대한약리학회지
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• 제23권2호
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• pp.101-111
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• 1987

중추신경계 특히 뇌내(腦內)의 무수카린성 콜린 수용체 (mAchR)에 대한 수용체 특성의 연구의 하나로, 물리 화학적 성상에 다른 두 종류의 콜린길항제를 사용하여 서로 다른 두 형태의 조직에서 약물의 작용양상 및 다른 약물과의 상호작용을 정초하였다. 실험동물로는 흰쥐를 일정기간 규정사료를 사육하였고, 사용한 Radioactive ligands는 $(^3H)$ QNB와 $(^3H)$ NMS였으며 그외에 다른 수종의 길항제 또는 효능제와의 치환작용을 brain homogenates와 intact brain cell aggregates에서 관찰하여 다음과 같은 결과를 얻었다. 1. $(^3H)$ QNB와 $(^3H)$ NMS는 모두 질량작용의 법칙에 비례하여 수용체와의 결합에서 높은 친화력과 포화를 보였으며 또한 높은 결합 능력을 나타내었다. 더욱이 homogenates 제제와 intact cell aggregates제제에서의 결과 사이에는 유사한 점이 많았다. 2. Homogenates제제를 사용한 실험에서, 제 3 급아민콜린길항제인 QNB, atropine과 scopolamine 또는 제 4 급 암모늄골린 길항제인 methylatropine과 methylscopolamine을 사용하여 위의 radioactive ligands와의 치환작용을 검토하였다. $(^3H)$ NMS 실험군에서는 제 3 급아민 및 제 4 급 암모늄길항제 모두가 구조의 구별없이 질량작용의 법칙에 따라 치환되었으나 $(^3H)$ QNB 실험군에서는 제 4급 암모늄콜린 길항제들을 단일성(unity)이 아닌 높고 낮은 두 종류의 친화도를 가진 결합부위의 양상을 나타내었다. 또 비특이성 콜린길항제인 pirenzepine을 사용한 실험군에서는 두 ligands을 모두 치환시켰고 서로 다른 결합부위가 있음을 보였다. 3. Intact cell aggregates 제제를 사용한 실험에서, $(^3H)$ NMS와 $(^3H)$ QNB 모두 homogenates 제제에서와 같은 양상의 반응을 보였다. 또 $(^3H)$ NMS를 radioligand로 하여 수종의 콜린길항제와 수종의 콜린 효능제를 사용하여 약물 상호작용으로 수용체의 성질을 검토하였다. 그 결과 콜린 길항제들은 질량작용의 법칙에 따라 치환되었으나 콜린 효능제 투여군에서는 높고 낮은 두 종류의 다른 친화력의 결항부위를 나타내었다. 4. 위의 실험의 결과로,(a) 친유성콜린 길항제인 $(^3H)$ QNB는 친수성 콜린길항제인 $(^3H)$ NMS보다 훨씬 높은 결합능력을 보였으며 이것으로 수용체 특히 mAchR의 존재 장소 또는 mAchR의 형상의 일부는 세포막 표면 뿐 아니라 세포막내의 어떤 부위와도 관계가 되는 것으로 간주되는데 이것이 $(^3H)$ QNB가 $(^3H)$ NMS보다 높은 최대 결합능력 $(B_{max})$을 나타낼 이유이다. (b) 두 종류의 다른 제제에서 우리는 같은 양상의 결과를 관찰하었기에 결점이 많은 homogenates 제제보다는 intact cell aggregates 제제를 수용체 연구에 대한 새로운 실험모형(experiment model)으로 사용할 수 있는 가능성을 제시하고자 한다.