• The Korean Journal of Physiology and Pharmacology
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• 제22권5호
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• pp.539-546
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• 2018

Botulinum toxin type A (BoNT/A) has been used therapeutically for various conditions including dystonia, cerebral palsy, wrinkle, hyperhidrosis and pain control. The substantia gelatinosa (SG) neurons of the trigeminal subnucleus caudalis (Vc) receive orofacial nociceptive information from primary afferents and transmit the information to higher brain center. Although many studies have shown the analgesic effects of BoNT/A, the effects of BoNT/A at the central nervous system and the action mechanism are not well understood. Therefore, the effects of BoNT/A on the spontaneous postsynaptic currents (sPSCs) in the SG neurons were investigated. In whole cell voltage clamp mode, the frequency of sPSCs was increased in 18 (37.5%) neurons, decreased in 5 (10.4%) neurons and not affected in 25 (52.1%) of 48 neurons tested by BoNT/A (3 nM). Similar proportions of frequency variation of sPSCs were observed in 1 and 10 nM BoNT/A and no significant differences were observed in the relative mean frequencies of sPSCs among 1-10 nM BoNT/A. BoNT/A-induced frequency increase of sPSCs was not affected by pretreated tetrodotoxin ($0.5{\mu}M$). In addition, the frequency of sIPSCs in the presence of CNQX ($10{\mu}M$) and AP5 ($20{\mu}M$) was increased in 10 (53%) neurons, decreased in 1 (5%) neuron and not affected in 8 (42%) of 19 neurons tested by BoNT/A (3 nM). These results demonstrate that BoNT/A increases the frequency of sIPSCs on SG neurons of the Vc at least partly and can provide an evidence for rapid action of BoNT/A at the central nervous system.

• 대한수의학회지
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• 제46권4호
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• pp.323-326
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• 2006

Suaeda (S.) asparagoides $M_{IQ}$, one of the halophyte groups, has been used as a folk remedy for digestive disturbances in Korea. However, its pharmacological activity on gastrointestinal motility has not been reported yet. In this study, the effects of this halophyte extracts with various solvent fractions (ethanol, hexane, chloroform, ethyl acetate, butanol, and water) on mice ileal spontaneous motility was examined. All solvent fractions at the concentration of $100{\mu}g/ml$ showed inhibitory actions on spontaneous motility of ileum with the potency order of water > 70% ethanol > hexane ${\gg}$ chloroform ${\geq}$ butanol ${\geq}$ ethyl acetate, respectively. In addition, the water fraction of extracts from S. asparagoides $M_{IQ}$ (WFSA) dose-dependently ($1-100{\mu}g/ml$) inhibited the amplitude of spontaneous phasic contraction and area under the contractile curve (AUC). The inhibitory effect of water fraction at the concentration of $10{\mu}g/ml$ was not affected by tetrodotoxin (TTX), $Na^+$ channel blocker ($1{\mu}M$), and $N^w$-nitro-L-arginine Methyl Ester (L-NAME), nitric oxide synthase inhibitor ($100{\mu}M$). However, cyclopiazonic acid (CPA, $10{\mu}M$), inhibitor of sarcoplasmic reticulum $Ca^{2+}$-ATPase, almost blocked the inhibitory effects of WFSA ($10{\mu}g/ml$) on the spontaneous phasic contraction of mouse ileum. But, CPA did not inhibit the lowering basal tone effects of WFSA. The result of this study showed that various extracts of S. asparagoides $M_{IQ}$ induce inhibitory effects on spontaneous contraction of mice ileal segments. More over, the polar solvent fractions were shown to be more potent than non-polar solvent fractions. The effects of S. asparagoides $M_{IQ}$ extracts are not mediated by nerve or nitric oxide. The inhibitory effects of WFSA at least partially mediated by sarcoplasmic reticulum $Ca^{2+}$-ATPase. However, further study is required to determine the exact pharmacological mechanisms of this halophyte on its gastrointestinal motility inhibitory effects.

• 대한본초학회지
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• 제26권4호
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• pp.75-81
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• 2011

Objectives : Magnoliae officinalis Cortex (MOC) has been used in traditional medicine for digestive diseases in Korea, China and Japan. However, Machili thunbergii Cortex (MTC) also has been used as a substitute of MOC in Korea sometimes. Thus, this study was carried out to investigate and compare the effects of MOC and MTC on intestinal motility of isolated small intestinal segments from ICR mouse. Methods : Changes in motility were recorded via isometric transducers connected to a data acquisition system and amplitude, frequency and area under the curve (AUC) of intestinal spontaneous phasic contraction were compared. Results : The MOC extracts ($1{\sim}{\mu}g/mL$) dose-dependently decreased both amplitudes and frequencies of the spontaneous phasic contraction, but not AUC. However, high concentration of MOC (100 ${\mu}g$/mL) evoked tonic contraction. And it was not inhibited by tetrodotoxin, a sodium channel blocker, and nifedipine, a L-type $Ca^{2+}$ channel antagonist. These results suggested that MOC (100 ${\mu}g$/mL)-induced tonic contraction is not mediated by nerve or L-type $Ca^{2+}$ channel. On the other hand, the MTC extracts dose-dependently inhibited amplitude and AUC, but not the frequency. Conclusions : Although both MOC and MTC affected intestinal motility, MOC is more effective on intestinal motility than MTC. And MOC has been used as a traditional medicine for a long time but not MTC. Thus, we suggested that MTC should not be used in Korea as a substitute of MOC and MOC might be useful traditional medicine for gastrointestinal disease. The mechanism of MOC is still remained to elucidate.

• 대한수의학회지
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• 제37권1호
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• pp.119-128
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• 1997

The relaxation of gastric fundus smooth muscles is the primary physiological event which induces the receptive relaxation of monogastric animals. L-arginine/Nitric oxide(L-arg/NO) system is known to mediate the inhibitory non-adrenergic non-cholinergic(NANC) neurotransmission in various tissues including gastrointestinal smooth muscles. The longitudinal smooth muscles of porcine gastric fundus showed fast relaxation during electrical field stimulation(EFS) and rebound contraction after EFS in NANC condition. So, the purpose of present study was elucidation of the neurotrasmitters related to the NANC relaxation and explanation of the relation between NANC relaxation and L-arg/NO system. The longitdinal smooth muscles of porcine gastric fundus were hung in the organ bath and under the presence of guanethidine($5{\times}10^{-5}M$), precontraction was induced by carbachol($1{\times}10^{-6}M$). The muscle responses to EFS and drugs were isomerically recorded. The rusults were summarized as follows. 1. The longtudinal muscles of porcine gastric fundus showed frequency-dependent relaxation and rebound contraction to electrical field stimulaton(1ms, 8V, 1~16Hz, 20sec, EFS). These responses were blocked by tetrodotoxin($1{\times}10^{-6}M$). 2. The relaxation and rebound contraction of the longitudinal muscles of porcine gastric fundus to EFS were inhibited by L-NAME($2{\times}10^{-5}M$). The inhibitory effect of L-NAME was antagonized by L-arginine($1{\times}10^{-3}M$), but not by D-arginine($1{\times}10^{-3}M$). 3. Exogenous NO($NaNO_2$, $1{\times}10^{-5}{\sim}1{\times}10^{-4}M$, pH=2.0) caused concentration-dependent relaxation as EFS did. 4. Methylene Blue($2{\times}10^{-5}M$), a soluble guanylate cyclase inhibitor, inhibited the relaxation and rebound contraction of the longitudinal muscles of porcine gastric fundus induced by EFS, but N-ethlmaleimide, a adenylate cyclase inhibitor, did not. 5. 8-Br-cGMP($1{\times}10^{-6}{\sim}3{\times}10^{-6}M$), permeable cGMP analogue, induced dose-dependent relaxation. but 8-Br-cAMP($1{\times}10^{-6}{\sim}3{\times}10^{-6}M$), permeable cAMP analogue, did not. Both did not evoked rebound contraction. 6. ${\alpha}$-chymotrypsin did not affect the relaxation of the longitudinal muscles of porcine gastric fundus. 7. Reactive blue 2($1{\times}10^{-4}M$, 40min) siginificantly inhibited the rebound contraction induced by EFS and inhibited contraction caused by exogenous ATP($1{\times}10^{-4}{\sim}1{\times}10^{-3}M$). These results suggests that NANC relaxation of the longitudinal muscles of porcine gastric fundus mainly mediated by NO and the rebound contraction is related to NO and other neurotransmitters.

• 대한수의학회지
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• 제35권3호
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• pp.459-469
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• 1995

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.

• 대한약리학회지
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• 제31권1호
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• pp.39-51
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• 1995

갑상선 기능 항진증 심장에서의 ${\beta}-adrenoceptor$의 역할은 잘 알려져 있으나 ${\alpha}-adrenoceptor$에 대해서는 알려져 있지 않은 바, 저자 등은 갑상선 기능 항진증 기니픽 심장의 유두근에서 일반 미세 전극과 이온-선택적 미세 전극을 이용하여 세포내 $Na^+$과 $H^+$ 활성도에 대한 phenylephrine의 영향을 연구하였다. Phenylephrine ($10^{-5}$ 또는 $3{\times}10^{-5}M$)에 의한 ${\alpha}_1-adrenoceptor$ 자극은 다양한 활동전위의 변동, 수축기 막전위의 과분극 ($1.5{\pm}0.1mM$), 세포내 활성도 증가 ($0.4{\pm}0.15mM$), 현저한 수축력 증가 ($220{\pm}15%$) 그리고 세포내 pH의 증가 ($0.06{\pm}0.002\;unit$)를 일으켰고, 이와 같은 변동이 prazosin과 atenolol에 의해 차단되었다. 그래서 이들 효과가 ${\alpha}_1-adrenoceptor$를 경유함을 알 수 있었고, 역시, 세포내 $Na^+$ 활성도와 수축력 증가 효과가 $Na^{+}-H^{+}$ 교환기 억제제인 ethylisopropylamiloride로 차단됨으로 보아 ${\alpha}_1-adrenoceptor$ 자극은 $Na^{+}-H^{+}$ 교환기를 자극하여 세포내 $a^{i}_{Na}$와 pH를 증가시킴을 시사한다. 이는 ${\alpha}_1-adrenoceptor$ 자극에 의한 세포내 $Na^+$감소와 초기 수축력 감소 효과를 나타내는 정상 기니픽 심장과는 매우 다른 결과로 갑상선 기능 항진증 심장에서 ${\alpha}_1-adrenoceptor$는 매우 중요한 기능을 갖고 있음을 의미한다.

• 한국임상수의학회지
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• 제6권2호
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• pp.227-259
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• 1989

Waterpepper is a weed which grows on damp soil. especially near swamps, and in shallow water of ponds and ditches. It Is widespread throughout the country In abundant colonies. In the present experiments, possible toxic effects of waterpepper were investigated in ruminants. Pour cows were fed waterpepper ad libitum or by force in the from of green forage, hay and/or powder, 8 goats were administered in the form of methanol extract, and 4 goats, crude juice, into the lumen. Clinical signs were examined as well as urinalysis, hematology, serum chemical analysis, pH/blood gas analysis and chclinesterase activities following administration of waterpepper. Six goats which were administered the methanol extract or crude juice were sacrificed for pathological examinations., In addition to the clini copathological examinations, the chemical constituents of waterpepper were qualitively analyzed from the methanol extract and the Effects of the waterpepper crude juice were examined on the motility of rabbit duodenum and uterus. It is revealed that waterpepper contains steroids, terpenoids, flavonoids, tannin and essential oils in the methanol extract and nitrates in the crude juice. The crude juice of waterpepper relaxed the rabbit uterine and duodenal smooth muscles. The constraction of duodenum by acetylcholine or BaCl$_2$ were partially inhibited by pretreatment of the crude juice. However, the relaxation of duodenum by the crude juice was not blocked by the pretreatments of phenoxybenzamlne, propranolol, cocaine, reserpine and tetrodotoxin. The constituents of waterpepper to evoke elaxation of duodenal smooth muscle were stable to heat. The cows administered waterpepper showed common clinical symptoms such as acrid expression, restlessness, dullness, inappetence, anorexia, severe diarrhea, mild bloat and left displacement of abomasum, while bloody feces was shown in a cow. The goats administered the mothanol extract showed common clinical signs such as acrid expression, restlessness, dullness, inappetence and soft feces, while bloody feces was shown in a goat, A goat adminstered the crude juice showde bloody feces and diarrhea. Respiratory rates and heart beats were increased along with diarrhea in the experimental cows. The erythrocyte counts and MCHC were decreased whereas PCV, MCV and neutrophils were increased in the cows administered waterpepper. In goats administered methanol extract, there were decreases in erythrocytes, PCV and hemoglobin content, and an increase in MCHC. The goats ingester with the crude juice showed negligible changes in hematologic values compared with control group which was administered the same amount of water instead of the crude juice. The contents of serum calcium, Inorganic phosphorus, magnesium, Iron, glucose, cholesterol, total protein, triglycerides and phospholipids were tended to decrease in cows. In goats serum iron, glucose, triglycerides, cholesterol, BUN and phospholipids content were decreased while the content of sodium and chloride were increased after administration of the methamol extract The goats ingested with the crude juice did not show significant changes in serum chemical analysis. Even though there were some pathological findings such as hyperemia in the small intestines and kidneys and swelling of liver parenchymal cells, the values of serum AST, ALT, LDH, alkaline phosphatase, total bilirubin and creatinine did not change significantly. While proteins, hemoglobin and blood were detected in the urine of cows, urine pH, ketone bodies, glucose, bilirubin and urobilinogen content were normal or undetected. There were no significant changes in pH/bolld gas analysis data of cows and cholinesterase activities of plasma and erythrocytes of cows and goats ingested with waterpepper or the methanol extract. It is concluded that waterpepper irritates the gatrointestinal system, causes abdominal pain, relaxes the gastrointestinal smooth muscle and dilatates blood vessels supplied to the system. The irritation and relaxation may lead to abnormal fermentation, maldigestion and malabsorption of nutrients and result in diarrhea, body feces, mild bloat and left displacement of abomasum.

• 대한약리학회지
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• 제32권3호
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• pp.301-309
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• 1996

흰쥐 해마(hippocampus)에서 norepinephrine(NE) 유리에 미치는 $A_{1}-adenosine$ 수용체의 post-receptor 기전에 $K^+$-통로가 관여하는지에 대한 지견을 얻고자 하여 $^3H-NE$로 평형시킨 해마 절편을 사용하여 adenosine의 $^3H-NE$ 유리에 미치는 $K^+$-통로 차단제의 영향을 관찰하였다. Adenosine$(1{\sim}30{\mu}M)$은 전기자극(3 Hz, 2 ms, 5 $VCm^{-1}$, 구형파)에 의한 NE 유리를 용량 의존적으로 감소시켰다. $K^+$-통로 차단제의 하나인 4-aminopyridine(4AP, $1{\sim}30{\mu}M$)은 자극에 의한 NE 유리를 용량 의존적으로 증가시켰으며 특히 10 및 $30{\mu}M$의 투여에 의해 기저 유리 또한 증가시켰다. 또 다른 $K^+$-통로 차단제인 tetraethylammonium(TEA, $1{\sim}10mM$) 역시 자극에 의한 NE 유리를 용량 의존적으로 증가시켰으나 이때 기저 유리에는 변화를 보이지 않았다. $K^+$-통로 차단제의 adenosine 효과에 미치는 실험에서는 adenosine에 의한 NE 유리 감소효과가 4AP $3{\mu}M$ 동시 투여에 의해 억제되었으며, 또한 1 mM TEA에 의하여는 영향을 받지않았으나 3 mM TEA 동시 투여에 의하여는 억제됨을 볼 수 있었다. 한편 $30\;{\mu}M$ 4AP 에 의한 NE 유리 증가효과는 $Ca^{++}$ 제거 영양액에서는 완전히 소실되었고 영양액 내의 $Ca^{++}$을 정상 농도의 1/4로 하였을때에는 NE 유리 억제가 어느정도 회복됨을 볼 수 있었으며 이때 기저 유리 또한 증가됨을 볼 수 있었다. l/4 $Ca^{++}$ 농도시에 4AP의 NE유리에 미치는 효과는 영양액내의 $Mg^{++}$을 4mM로 올렸을때 크게 억제되었으며 $0.3\;{\mu}M$ tetrodotoxin(TTX) 동시 투여에 의해 완전히 차단됨을 볼 수 있었다. TEA 10mM에 의한 NE 유리 증가효과 역시 $Ca^{++}$ 제거 영양액에서 완전히 소실되었고 이 또한 1/4 $Ca^{++}$ 영양액에 의하여는 회복됨을 볼 수 있었으며 $Mg^{++}$ 증가 영양액에서는 억제, TTX 동시 투여시에는 완전히 소실되었다. 이상의 실험결과로 흰쥐 해마에서 $A_1-adenosine$ 수용체를 통한 adenosine의 NE 유리 감소는 TEA 및 4AP에 예민한 $K^+$-통로가 관여하고 여기에는 세포외액의 Ca^{++}의 농도가 중요한 인자의 하나로 관여 하는 것으로 사료된다.

• 대한약리학회지
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• 제16권1호
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• pp.25-34
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• 1980

기니픽 요관(尿管)을 in vitro로 전기자극(電氣刺戟)할 때 일어나는 ON response와 전기자극(電氣刺戟) 중지시(中止時)에도 spike성(性) OFF response가 일어남을 관찰(觀察)하고 이에 대한 특성(特性)을 검토하였기에 요약(要約)한다. 1. 기니픽 요관(尿管)을 in vitro로 5초간 연속(連續)으로 전기자극(電氣刺戟)(EFS)시(時) 두 수축(수축)을 일으켰다. EFS 시작 후 $0.1{\sim}0.3$ sec에 ON response가 일어나고 EFS 종료 후 $0.2{\sim}0.1$ sec에 OFF response가 야기(惹起)되었다. OFF response를 일으키는 연속(連續) 자극(刺戟)에는 ON response에 비(比)하여 긴 puise duration과 높은 frequency가 요구(要求)되었다. 2. Muscle chamber내(內)의 온도(溫度)를 $37^{\circ}C$에서 $22^{\circ}C$로 감소(減少)시켰을 때는 ON의 진폭(振幅)은 감소(減少)하거나 경미(輕微)하였고 OFF는 증가(增加)되었으며, 온도(溫度) $42^{\circ}C$에서는 OFF가 감소(減少) 또는 소실(消失)되던 것이 $K^+$-free solution 에서는 덜 감소(減少)되었다. ON 및 OFF response의 latency는 온도(溫度)의 하강(下降)으로 증가(增加)되었고 그 정도(程度)는 양자(兩者)가 비슷하였다. 3. 저(低) $Ca^{++}$ 농도(濃度), 2mM EDTA, 1mM $Mn^{++}$ 및 $1{\mu}M$ verapamil 등(等)에 의해 반복(反復) 자극시(刺戟時) ON은 쉽게 소실(消失)되고 OFF는 남아 있었다. 4. $10{\mu}M$ theophylline은 ON 진폭(振幅)은 감소(減少)시키지 않고 OFF response만 소실(消失)시키고, 1mM sodium nitroprusside는 OFF에는 작용(作用)하지 않고 ON 진폭(振幅)을 우선적(優先的)으로 감소(減少)시켰다. 5. ON 및 OFF response는 TTX, atropine, phentolamine 등(等) 신경성봉쇄약물(神經性封鎖藥物)에 의하여 영향(影響)을 받지 아니할 뿐 아니라 pyrilamine이나 methysergide에 의하여도 작용(作用)을 받지 아니하였다. 6. 50% 저(低) sodium에 의하여는 ON 및 OFF response는 影響(影響)을 받지 아니하였고 ON과 OFF 사이의 긴장(긴장)은 저(低) sodium에서 증가(增加)되었고, voltage 감소(減少)에 의하여 원상(原狀)으로 회복(恢復)되었다. 이상(以上)의 성적(成績)으로 보아 ON response는 세포(細胞) 외(外)의 free $Ca^{++}$에 의존(依存)하고 OFF response는 세포내(細胞內)에 저장되거나 막(膜)에 bound된 $Ca^{++}$에 더 의존(依存)하며, 생물학적(生物學的) 현상(現象)에 의(依)하여 나타나는 반응(反應)일 것으로 사료(思料)되는 바이다.