• Biomolecules & Therapeutics
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• v.2 no.4
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• pp.336-342
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• 1994

The general pharmacological properties of EPO were investigated in various animals administering intravenously and in vitro system. The results were as follows. 1. Central nervous system: EPO at doses of 70, 700, 7000 U/kg showed no effect In mice on general behavior, on strychnine- and pentetrazol-induced convulsion and on acetic acid-induced writhing syndrome. The hexobarbital-induced sleeping time in mice was slightly reduced by EPO at a dose of 7000 U/kg but did not change at doses of 70, 700 U/kg. The body temperature in rats was slightly decreased by EPO at doses of 700, 7,000 U/kg but the change was in normal physiological range. 2. Respiratory and cardiovascular system: EPO showed no effect on respiratory rate, blood pressure, heart rate, femoral blood flow, and electrocardiogram in anesthetized dogs at doses of 70, 700, 7000 U/kg. 3. Smooth muscle: EPO at concentrations of 70, 700 U/ml had no effect on the contractile response of isolated guinea pig ileum to histamine and acetylcholine. 4. Water and electrolytes excretion: EPO at dose above 700 U/kg increased urine volume in rats but did not affect the concentrations of $Na^{+},\;K^{+},\;Cl^{-}$ in urine. 5. Gastrointestinal system: EPO(70, 700, 7000 U/kg) had no effect on the intestinal charcoal meal propulsion 6. Blood coagulation system: The administration of EPO(70, 700, 7000 U/kg) had no effect on the plasma prothrombin time(PT) and activated partial thromboplastin time(APTT) in mice. Platelet aggregation induced by ADP and collagen was not influenced by EPO(70 U/ml, 700 U/ml). The overall results obtained indicated that EPO exerts almost no serious pharmacological effect even at a 100-fold clinical dose(7000 U/kg).

• Experimental and Molecular Medicine
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• v.50 no.12
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• pp.11.1-11.9
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• 2018

Estrogen has diverse effects on cardiovascular function, including regulation of the contractile response to vasoactive substances such as serotonin. The serotonin system recently emerged as an important player in the regulation of vascular tone in humans. However, hyperreactivity to serotonin appears to be a critical factor for the pathophysiology of hypertension. In this study, we examined the modulatory mechanisms of estrogen in serotonin-induced vasoconstriction by using a combinatory approach of isometric tension measurements, molecular biology, and patch-clamp techniques. $17{\beta}$-Estradiol (E2) elicited a significant and concentration-dependent relaxation of serotonin-induced contraction in deendothelialized aortic strips isolated from male rats. E2 triggered a relaxation of serotonin-induced contraction even in the presence of tamoxifen, an estrogen receptor antagonist, suggesting that E2-induced changes are not mediated by estrogen receptor. Patch-clamp studies in rat arterial myocytes showed that E2 prevented Kv channel inhibition induced by serotonin. Serotonin increased Src activation in arterial smooth muscle required for contraction, which was significantly inhibited by E2. The estrogen receptor-independent inhibition of Src by E2 was confirmed in HEK293T cells that do not express estrogen receptor. Taken together, these results suggest that estrogen exerts vasodilatory effects on serotonin-precontracted arteries via Src, implying a critical role for estrogen in the prevention of vascular hyperreactivity to serotonin.

• The Korean Journal of Physiology and Pharmacology
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• v.1 no.5
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• pp.529-535
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• 1997

The present study was aimed at investigating whether the vascular calcium regulation is altered in hypertension. Two-kidney, one clip (2K1C) and deoxycorticosterone acetate (DOCA)-salt hypertension were made in rats, and their thoracic aortae were taken 4 weeks later. The isometric contractile response and calcium uptake of the endothelium-denuded aortic preparations were determined. Caffeine ($0.1{\sim}35\;mmol/L$) induced a greater contraction in 2K1C and DOCA-salt hypertension than in normotensive control. When the vascular calcium store was functionally-depleted by a repeated exposure to caffeine, it took longer to reload the store and to resume the initial contraction force in response to caffeine in both 2K1C and DOCA-salt hypertension. The vascular $^{45}Ca$ uptake following the functional depletion of the cellular store was also greater in both models of hypertension than in control. Ryanodine, calcium channel activator of the sarcoplasmic reticulum, attenuated the restoration of caffeine-induced vascular contraction, which was not affected by either 2K1C or DOCA-salt hypertension. Nifedipine, an L-type $Ca^{2+}$ channel blocker, attenuated the restoration of caffeine-induced contraction, which was not affected by DOCA-salt hypertension, but was more pronounced in 2K1C hypertension. Nifedipine also diminished the vascular $^{45}Ca$ uptake, which was not affected by DOCA-salt hypertension, but was more pronounced in 2K1C hypertension. Ouabain, a $Na^+,\;K^+-ATPase$ inhibitor, increased the caffeine-induced contraction by a similar magnitude in control and 2K1C hypertension, which was, however, markedly attenuated in DOCA-salt hypertension. Ouabain enhanced the vascular $^{45}Ca$ uptake, the degree of which was not affected by 2K1C hypertension, but was markedly attenuated in DOCA-salt hypertension compared with that in control. Cyclopiazonic acid, a selective inhibitor of $Ca^{2+}-ATPase$ of the sarcoplasmic reticulum, attenuated the restoration of caffeine-induced contraction, which was not affected by 2K1C hypertension, but was more marked in DOCA-salt hypertension. These results suggest that the increased vascular calcium storage may be attributed to an enhanced calcium influx in 2K1C hypertension, and to an impaired $Na^+-K^+$ pump activity of the cell membrane and subsequently increased calcium pump activity of the cellular store in DOCA-salt hypertension.

• The Korean Journal of Physiology and Pharmacology
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• v.4 no.3
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• pp.185-195
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• 2000

Vascular diseases are significant complications of diabetes mellitus (DM), and the endothelial cells may play a pivotal role in the development of vascular disease in DM. Endothelin-1 (ET-1) released from endothelium is a potent vasoconstrictor peptide and circulating level of ET-1 is increased in a variety of disease states. The purpose of this study was to determine the changes of responsiveness to ET-1 in DM, and we experimented on the changes in the ET-1-induced contraction, levels of nitrite and lipid peroxidation, and ET-1 immunoreactivity in aorta from streptozotocin-induced DM rats. DM was induced by single injection of streptozotocin (55 mg/kg, i.p.). The immunoreactive ET-1 levels in endothelial layer of thoracic aorta were much higher in DM rats than control rats. Nitrite in tissue homogenate was decreased and plasma nitrite was increased in DM rats. Malondialdehyde (MDA) was significantly increased in DM rats and cGMP was not significantly different between control and DM rats. ET-1 produced concentration- dependent contractile responses that are significantly attenuated in DM rats compared to controls. In the presence of selective $ET_A$ receptor antagonist BQ610, the maximum contraction was decreased and the concentration ratios for BQ610 yielded $pA_2$ values of 7.3 (slope, 0.65) in control rats, whereas BQ610 had no antagonistic effect on ET-1-induced contraction in DM rats. However, pretreatment with BQ788, an $ET_B$ receptor antagonist, maximum response was decreased and the dose-response curves for ET-1 were shifted to the right in both groups and $pA_2$ values were 7.9 and 7.7 (slope, 1.05 in control and DM rats), respectively. IRL 1620 and sarafotoxin S6c, $ET_B$ agonists, induced relaxation in control rats but not in DM rats. These results indicate that endothelial cell dysfunction and enhanced immunoreactivity of ET-1 have been found in DM rat and ET-1-induced contraction was attenuated in DM rat. These attenuated responses might be at least in part caused by the alteration of $ET_A$ receptor properties (e.g. desensitization), and partly related with an alteration in intracellular mechanism for contraction to ET-1.

• The Journal of Internal Korean Medicine
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• v.19 no.1
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• pp.385-408
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• 1998

Pyeongpaesan (平肺散) has been used in Korea for many centuries as a treatment for respiratory disease. The effect of Pyeongpaesan (平肺散) on tracheal smooth muscle is not known. The purpose of the present study is to determine the effect of Pyeongpaesan (平肺散) on histamine and acetylcholine induced tracheal smooth muscle contraction in rats and guinea pigs. Guinea pig (500 g, male) and Sprague Dawley rats (200 g, male) were killed by $CO_2$ exposure and a segment (8-10 mm) of the thoracic trachea from each rat and guinea pig was cut into equal segments and mounted 'in pairs' in a tissue bath. Contractile force was measured with force displacement transducers under 0.5 g loading tension. The dose of histamine (His) and acetylcholine (Ach) which evoked 50% of maximal response ($ED_{50}$) was obtained from cumulative dose response curves for histamine and acetylcholine $(10^{-7}{\sim}10^{-4}M)$. Contractions evoked by His $(ED_{50})$ and Ach $(ED_{50})$ were inhibited significantly by Pyeongpaesan (平肺散). In guinea pig tracheal smooth muscle, the mean percent inhibition of acetylcholine induced contraction was 13.5% (p<0.05) after $10{\mu}l/ml$ Pyeongpaesan (平肺散), $64.6\(p<0.01)\;after\;30{\mu}l/ml$ Pyeongpaesan (平肺散), and $92.8\(p<0.01)\;after\;100{\mu}l/ml$ Pyeongpaesan (平肺散). In rat tracheal smooth muscle, the mean percent inhibition of acetylcholine induced contraction was $60.9\(p<0.01)\;after\;30{\mu}l/ml$ Pyeongpaesan (平肺散), and $91.2\(p<0.01)\;after\;100{\mu}l/ml$ Pyeongpaesan (平肺散). Also, in guinea pig tracheal smooth muscle, the mean percent inhibition of histamine induced contraction was $104.8\(p<0.01)\;after\;30{\mu}l/ml$ Pyeongpaesan (平肺散) and $142.3\(p<0.01)\;after\;100{\mu}l/ml$ Pyeongpaesan (平肺散). In rat tracheal smooth muscle, the mean percent inhibition of histamine induced contraction was $63.7\(p<0.01)\;after\;30{\mu}l/ml$ Pyeongpaesan (平肺散), and $107.5\(p<0.01)\;after\;100{\mu}l/ml$ Pyeongpaesan (平肺散). Propranolol $(10^{-7}M)$ slightly but significantly attenuated the inhibitory effects of Pyeongpaesan (平肺散). Following treatment with propranolol, the mean percent inhibition caused by $100{\mu}l/ml$ Pyeongpaesan (平肺散) fell to 15.7% (p<0.05) in guinea pig induced by acetylcholine contraction and the mean percent inhibition caused by $100{\mu}l/ml$ Pyeongpaesan (平肺散) fell to 22.3% (p<0.05) in guinea pig induced by histamine contraction and by $100{\mu}l/ml$ Pyeongpaesan (平肺散) fell to 28.7% (p<0.01) in rat induced by histamine contraction. Indomethacin and methylene blue $(10^{-7}\;M)$ did not significantly alter the inhibitory effect of Pyeongpaesan (平肺散). Also, I could find the effects of Pyeongpaesan (平肺散) and Pyeongpaesanga (平肺散加) morphine on the tracheal smooth muscle in guinea pig and rat did not change significantly. These results indicate that Pyeongpaesan. (平肺散) can relax histamine and acetylcholine-induced contraction of guinea pig and rat tracheal smooth muscle, and that this inhibition involves sympathetic effects and the release of cyclooxygenase products.

• Journal of Physiology & Pathology in Korean Medicine
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• v.17 no.2
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• pp.493-498
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• 2003

Paeoniae Radix, Glycyrrhizae Radix and Jakyakgamchotang have been used in Oriental Medicine for many centuries as a treatment for various disease. The purpose of the present study is to determine the effect of Paeoniae Radix, Glycyrrhizae Radix and Jakyakgamchotang on narepinephrine(NE) induced contraction of isolated rabbit femoral artery. Rabbits (2.0kg, female) were killed by CO₂ exposure and a segment (8-10mm) of the aortic ring from each rabbit was cut into equal segments and mounted 'in pairs' in a tissue bath. Contractile force was measured with force displacement transducers under 1.5g loading tension. The dose of NE which evoked 50% of maximal response (ED/sub 50/) was obtained from cumulative dose response curves for NE (10/sup -7/～10/sup -4/M). Contractions evoked by NE (ED/sub 50/) were inhibited significantly by Paeoniae Radix, Glycyrrhizae Radix and Jakyakgamcho-tang. The mean percent inhibition of NE induced contraction was 83.9% (p<0.01) after 150㎕/㎖ Paeoniae Radix, 101.1 %(p<0.01) after 150 ㎕/㎖, Glycyrrhizae Radix and 107.3%(p<0.01) after 150㎕/㎖ Jakyakgamcho-tang, Indomethacin slightly but significantly attenuated the inhibitory effects of Paeoniae Radix. Following treatment with indomethacin, the mean percent inhibition caused by 150㎕/㎖ Paeoniae Radix fell to 16.4% in femoral artery induced by NE contraction. Propranolol, ODQ, and L-NNA did not significantly alter the inhibitory effect of Paeoniae Radix. ODQ slightly but significantly attenuated the inhibitory effects of Glycyrrhizae Radix. Following treatment with ODQ, the mean percent inhibition caused by 150㎕/㎖ Glycyrrhizae Radix fell to 13.0% in femoral artery induced by NE contraction. Propranolol, indomethacin and L-NNA did not significantly alter the inhibitory effect of Glycyrrhizae Radix. L-NNA slightly but significantly attenuated the inhibitory effects of Jakyakgamchotang. Following treatment with L-NNA, the mean percent inhibition caused by 150㎕/㎖ Jakyakgamchotang fell to 13.8% in femoral artery induced by NE contraction. Propranolol, ODQ and indomethacin did not significantly alter the inhibitory effect of Jakyakgamcho-tang. These results indicate that Paeoniae Radix, Glycyrrhizae Radix and Jakyakgamcho-tang can relax NE induced contraction of the isolated rabbit femoral artery, and that this inhibition related to nitric oxide.

• Korean Journal of Acupuncture
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• v.18 no.1
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• pp.157-164
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• 2001

Radix Asteris has been used in Korea for many centuries as a treatment for respiratory disease. The effect of Radix Asteris on tracheal smooth muscle is not known. The purpose of the present study is to determine the effect of Radix Asteris on histamine induced tracheal smooth muscle contraction in rats and guinea pigs. Guinea pig(500g, male) and Sprague Dawley rats (250g, male) were killed by $CO_2$ exposure and a segment (8-10mm) of the thoracic trachea from each rat and guinea pig was cut into equal segments and mounted 'in pairs' in a tissue bath. Contractile force was measured with force displacement transducers under 0.5g loading tension. The dose of histamine (His) which evoked 50% of maximal response ($ED_{50}$) was obtained from cumulative dose response curves for histamine $(10^{-7}{\sim}10^{-4}M)$. Contractions evoked by His ($ED_{50}$) were inhibited significantly by Radix Asteris. In guinea pig tracheal smooth muscle, the mean percent inhibition of histamine induced contraction was 120.5% (p<0.01) after $100{\mu}l/ml$ Radix Asteris. In rat tracheal smooth muscle, the mean percent inhibition of histamine induced contraction was 135.4% (p<0.01) after $100{\mu}l/ml$ Radix Asteris. Propranolol $(10^{-7}M)$ slightly but significantly attenuated the inhibitory effects of Radix Asteris. Following treatment with propranolol, the mean percent inhibition caused by $100{\mu}l/ml$ Radix Asteris fell to 44.6% in guinea pig induced by histamine contraction and by $100{\mu}l/ml$ Radix Asteris fell to 18.7% (p<0.05) in rat induced by histamine contraction. Indomethacin and methylene blue$(10^{-7}M)$ did not significantly alter the inhibitory effect of Radix Asteris. These results indicate that Radix Asteris can relax histamine induced contraction of guinea pig and rat tracheal smooth muscle, and that this inhibition involves sympathetic effects.

• Journal of Life Science
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• v.22 no.3
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• pp.387-397
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• 2012

In this study, we focused on myomoduline A (MMA) released from the central nervous system of Aplysia kurodai. The primary structure of MMA is Pro-Met-Ser-Met-Leu-Arg-Leu-$NH_2$. This peptide is the same as that of the myomodulin family peptide found in other mollusks. The purified MMA showed a modulating activity of phasic contraction on the anterior byssus retractor muscle (ABRM) of Mytilus edulis. In order to study the relationship between the structure and biological activity of MMA, we synthesized MMA, Des[$Pro^1$]-MMA, Des[$Pro^1,Met^2$]-MMA, Des[$Pro^1,Met^2,Ser^3$]-MMA, and MME. The amino acid sequences of Des[$Pro^1$]-MMA, Des[$Pro^1,Met^2$]-MMA, and Des[$Pro^1,Met^2,Ser^3$]-MMA were Met-Ser-Met-Leu-Arg-Leu-$NH_2$, Ser-Met-Leu-Arg-Leu-$NH_2$, and Met-Leu-Arg-Leu-$NH_2$, respectively. MMA and synthetic peptides were tested on ABRM in M. edulis as well as muscle preparations in Achatina fulica. At $1{\times}10^{-8}$ M or lower, MMA showed a potentiating effect on phasic contraction of the ABRM, but this peptide had an inhibitory effect at $1{\times}10^{-6}$ M or higher. Both MMA and its analogs stimulated a contractile response on the crop and a relaxed catch-relaxing response on the penial retractor muscle of A. fulica. These results suggest that Met-Leu-Arg-Leu-$NH_2$ in MMA is the minimum structure required for the regulation of the contraction of ABRM, as well as the reproductive and digestive activities of mollusks.

• Archives of Pharmacal Research
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• v.24 no.6
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• pp.546-551
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• 2001

We have previously shown that, in circular muscle cells of the lower esophageal sphincter (LES) isolated by enzymatic digestion, contraction in response to maximally effective doses of acetylcholine (ACh) or Inositol Triphosphate ($IP_3$) depends on the release of $Ca^{2+}$ from intracellular stores and activation of a $Ca6{2+}$-calmodulin (CaM)-dependent pathway. On the contrary, maintenance of LES tone, and response to low doses of ACh or $IP_3$ depend on a protein kinase C (PKC) mediated pathway. In the present investigation, we have examined requirements for $Ca6{2+}$ regulation of the interaction between CaM- and PKC-dependent pathways in LES contraction. Thapsigargin (TG) treatment for 30 min dose dependently reduced ACh-induced contraction of permeable LES cells in free $Ca6{2+}$ medium. ACh-induced contraction following the low level of reduction of $Ca6{2+}$ stores by a low dose of TG ($10^{-9}{\;}M$) was blocked by the CaM antagonist, CCS9343B but not by the PKC antagonists chelerythrine or H7, indicating that the contraction is CaM-dependent. After maximal reduction in intracellular $Ca{2+}$ from $Ca6{2+}$stores by TG ($10^{-6}{\;}M$), ACh-induced contraction was blocked by chelerythrine or H7, but not by CCS9343B, indicating that it is PKC-dependent. In normal $Ca^{2+}$medium, the contraction by ACh after TG ($10^{-9}{\;}M$) treatment was also CaM-dependent, whereas the contraction by ACh after TG ($10^{-9}{\;}M$) treatment was PKC-dependent. We examined whether PKC activation was inhibited by activated CaM. CCS 7343B Inhibited the CaM-induced contraction, but did not inhibit the DAC-induced contraction. CaM inhibited the DAC-induced contraction in the presence of CCS 9343B. This inhibition by CaM was $Ca{2+}$dependent. These data are consistent with the view that the switch from a PKC-dependent pathway to a CaM dependent pathway can occur and can be regulated by cytosolic $Ca{2+}$ in the LES.

• The Korean Journal of Pharmacology
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• v.10 no.1 s.15
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• pp.31-40
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• 1974

1. The authors investigated the effects of $PGE_1$ on the action of sympathomimetics in the vas deferens of guinea-pig, comparing with those in the rat vas deferens, and also the action of $PGE_1$ on the motility of nerve-free smooth muscle of chick amnion. 2. In the isolated guinea-pig vas deferens, the actions of phenylephrine and norepinephrine were much potentiated by pretreatment with $PGE_1$. Futher, in the isolated hypogastric nerve-vas deferens preparation of guinea-pig, effects of phenylephrine, norepinephrine and tyramine on the contractile response of vas to the hypogastric nerve stimulation and to the transmural stimulation were also augumented especially in tension by $PGE_1$-pretreatment. 3. In the isolated hypogastric nerve-vas preparation of rat, both contractile responses to hypogastric nerve and transmural stimulation were slowly reduced by treatment with $PGE_1$ and the potentiated effect of phenylephrine or norepinephrine was not observed in spite of pretreatment with $PGE_1$. 4. The actions of phenylephrine and norepinephrine on the denervated vas deferens of guinea-pig were also enhanced by $PGE_1$ as it were in the intact vas deferens, but there was no significant effect by $PGE_1$ on the action of norepinephrine in the denervated rat vas deferens. 5. $PGE_1$ in low concentration $(10^{-8}g/ml)$ did not affect the spontaneous motility of nerve-free smooth muscle of chick amnion ($9{\sim}11$ th day incubated chick), but in large concentration $(5{\times}10^{-8}g/ml)$ it caused irregular and slightly inhibitory movement. Pretreatment with $PGE_1$ on chick amnion did not exert any change on the action of phenylephrine applied. However, the stimulatory action of physostigmine on the chick amnion was a little antagonized by the low concentration of $PGE_1$. 6. It might be summarized that there is species difference between the actions of $PGE_1$ on the vas deferens of guinea-pig and that of rat, and the action of $PGE_1$ on the guinea-pig vas deferens might be mediated by the other mechanism rather than by direct action on the vas musculature.