Insamjungchuntang has been used in Korea for many centuries as a treatment for respiratory disease. The effect of Insamjungchuntang on tracheal smooth muscle is not known. The purpose of the present study is to determine the effect of Insamjungchuntang 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 Insamjungchuntang. In guinea pig tracheal smooth muscle, the mean percent inhibition of acetylcholine induced contraction was $38.58\(p<0.05)\;after\;10{\mu}l/ml$ Insamjungchuntang, $90.75\(p<0.01)\;after\;30{\mu}l/ml$. Insamjungchuntang and $133.17\(p<0.01)\;after\;100{\mu}l/ml$ Insamjungchuntang. In rat tracheal smooth muscle, the mean percent inhibition of acetylcholine induced contraction was $10.0\(p<0.05)\;after\;10{\mu}l/ml$ Insamjungchuntang, $80.71\(p<0.01)\;after\;30{\mu}/ml$ Insamjungchuntang and $118.29\(p<0.01)\;after\;100{\mu}l/ml$ Insamjungchuntang. Also, in guinea pig tracheal smooth muscle, the mean percent inhibition of histamine induced contraction was $45.5\(p<0.01)\;after\;10{\mu}l/ml$ lnsamjungchuntang, and $93.17\(p<0.01)\;after\;30{\mu}l/ml$. lnsamjungchuntang $134.50\(p<0.01)\;after\;100{\mu}l/ml$ Insamjungchuntang. In rat tracheal smooth muscle, the mean percent inhibition of histamine induced contraction was $37.83\(p<0.01)\;after\;10{\mu}l/ml$ lnsamjungchuntang, $90.5\(p<0.01)\;after\;30{\mu}l/ml$ Insamjungchuntang and $135.17\(p<0.01)\;after\;100{\mu}l/ml$ Insamjungchuntang. Propranolol $(10^{-7}\;M)$ slightly but significantly attenuated the inhibitory effects of Insamjungchuntang. Following treatment with propranolol, the mean percent inhibition caused by $100{\mu}l/ml$. Insamjungchuntang fell to 46.42% in guinea pig induced by acetylcholine contraction and by $100{\mu}l/ml$ Insamjungchuntang fell to 5.43% (p<0.05) in rat induced by acetylcholine contraction and the mean percent inhibition caused by $100{\mu}l/ml$ Insamjungchuntang fell to 49.0% in guinea pig induced by histamine contraction and by $100{\mu}l/ml$ Insamjungchuntang fell to 48.6% (p<0.05) in rat induced by histamine contraction. Indomethacin and methylene blue $(10^{-7}\;M)$ did not significantly alter the inhibitory effect of lnsamjungchuntang. Also, I could find the effects of lnsamjungchuntang and Insamjungchuntanggamorphine on the tracheal smooth muscle in guinea pig and rat did not change significantly. These results indicate that Insamjungchuntang can relax histamine and acetylcholine-induced contraction of guinea pig and rat tracheal smooth muscle, and that this inhibition involves sympathetic effects.
This study was conducted to investigate antigenic potential of DA-3030, a recombinant human granulocyte-colony stimulating factor, in guinea pigs and mice. In the active systemic anaphylaxis test, the guinea pigs sensitized with 1.25 or 12.5 $\mu\textrm{g}$/head of DA-3030 alone did not show any anaphylactic reaction. In the homologous passive cutaneous anaphylaxis reaction, anti-DA-3030 antibody was not detected in guinea pigs sensitized with 1.25 or 12.5 $\mu\textrm{g}$/head of DA-3030 alone. On the other hand, the guinea pigs sensitized with 12.5 $\mu\textrm{g}$/heed of DA-3030 incorporated in Freund's complete adjuvant(FCA) or 1 mg/head of ovalbumin incorporated in FCA showed anaphylactic reaction. Anti-DA-3030 antibody was also detected in those guinea pigs. In immunodiffusion test using the sera sensitized with DA-3030 incorporated in FCA, precipitating antibodies were detected only in the sera sensitized with DA-3030 or DA-3030 incorporated in FCA showed. In 24-hour heterologous PCA reaction with sera of C57BL/6 mice immunized with 1.25 or 12.5 $\mu\textrm{g}$/head of DA-3030 alone, none of the sera showed positive reaction. But sera of the animals immunized with 12.5 $\mu\textrm{g}$/head of DA-3030 incorporated in aluminum hydroxide gel(Alum) or 5 $\mu\textrm{g}$/head of ovalbumin incorporated in alum showed positive PCA reaction. DA-3030 did not cause anaphylactic shock or passive cutaneous anaphylaxis in guinea pigs and mice when given alone although DA-3030 incorporated in FCA or Alum induced anaphylactic shock and passive cutaneous anaphylaxis. From these results, it may be concluded the DA-3030 does not induce systemic allergic reaction when administered alone in its clinical use.
An, Dong-jun;Kim, Byoung-han;Jung, Byeong-yeal;Yi, Chul-hyun;Jeon, Woo-jin;Lee, Pil-soo;Chung, Gab-soo
Korean Journal of Veterinary Research
/
v.45
no.2
/
pp.215-221
/
2005
Canine coronavirus (CCV) causes a mild gastroenteritis in dogs. The virus is highly contagious. Although the virus was isolated more than thirty years ago, canine coronavirus infection continues to be a widespread problem. Mixed infections with both CCV and canine parvovirus (CPV) are common. Four kinds of commercial killed CCV vaccines are available in Korea. All the commercial vaccines should pass the National Assay for Veterinary Biologicals prior to release. For the potency test of CCV vaccine, it is necessary to use CCV antibody free dogs. The test requires not only kennels but high cost. To develop easy, efficient and economic potency test method for killed CCV vaccine using laboratory animals, a series of experiments with rabbits and guinea pigs were carried out in this study. In the preliminary test, the guinea pigs showed better immune responses than rabbits. The guinea pig was also easy to manage. So guinea pig was selected for the potency test animals. When the guinea pigs were inoculated twice with one dose of vaccine intramuscuarly each, slower and a little lower SN antibody titers were induced in guinea pigs than in dogs (about 2 kg body weight Beagle strain) given the same posology as guinea pigs'. It was concluded that guinea pigs could be substituted for dogs in the potency test of killed CCV vaccine.
The purpose of this study was to investigate the whitening effect of Angelicae dahuricae Radix water extract (ADWE) by its application on artificial tanning spots which were induced by 1,500 $mJ/cm^2$ of UVB radiation on the backs of brown guinea pigs weighing approximately 450~500 g. Thirty ${\mu}l$ of ADWE, at each application, were applied twice a day, 5 days a week, for 8 weeks to the guinea pigs. The artificial tanning spots were divided into 3 groups which were vehicle control group [propylene glycol: ethanol: water (5:3:2)], positive control group (2% hydroquinone) and experimental group (2% ADWE). The visible whitening effect and changes in melanin index were evaluated once a week. On completion of the experiment, the animals were sacrificed under anesthetization, and the artificial tanning spots were excised by biopsy punch and stained with Hematoxylin and Eosin (H&E) to observe histological change and also stained with Fontana-Masson's silver (F-M) and S-100 to observe melanin pigmentation and melanocytes. In the gross observation, the experimental group showed higher pigmentation than the positive control group but lower pigmentation than the vehicle control group. Histological observation confirmed that ADWE had a positive whitening effect by showing a lower distribution of melanin and melanocytes in the epidermis of experimental group than in the vehicle control group.
Na Kyung-Sang;Kwon Eui-Kwang;Soo Yeung-Hyung;Suh Eun-Mi;Han Jong-Hyun
Korean Journal of Acupuncture
/
v.18
no.1
/
pp.157-164
/
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.
To evaluate immunotoxicity of skin decontamination kit(SDK) newly-developed in Agency for Defense Development(ADD), delayed contact hypersensitivity (maximization) test and passive cutaneous anaphylaxis(PCA) test of SDK were performed and the results were compared with those of M 291. In maximization test, sensitization reaction was induced by id injection (2.5 mg / 0.1 $\textrm{m}{\ell}$/ guinea pig or 2.5 mg+CFA/0.1 $\textrm{m}{\ell}$/guinea pig) and topical application (2.5 mg/$\textrm{m}{\ell}$/guinea pig) with SDK or M291 at an interval of 1 week, and 2 weeks later, challenged by topical application with 25 mg/$\textrm{m}{\ell}$/guinea pig. SDK and M291 did not induce any reactions, showing 0 point of sensitization score and 0% of sensitization rate. In conclusion, it is suggested that SDK and M291 do not induce delayed contact hypersensitivity. In PCA test, rats were administered id with mouse anti-SDK serum and challenged iv with a mixture of antigen SDK and Evan's blue. SDK did not induce blue spots at the injection sites of both high (2.5 mg/mouse) and low (1.25 mg/mouse) dose-induced antisera. In contrast, BSA, positive control produced spots larger than 5 mm in diameter at the injection sites of BSA-induced antiserum up to $2^2$ ~ $2^4$dilution. In conclusion, it is suggested that SDK do not induce IgE production and is not a PCA-reaction inducer.
To investigate the possible involvement of outward potassium ($K^+$) currents in nitric oxide-induced relaxation in intestinal smooth muscle, we used whole-cell patch clamp technique in freshly dispersed guinea-pig ileum longitudinal smooth muscle cells. When cells were held at -60 mV and depolarized from -40 mV to -50 mV in 10 mV increments, sustained outward $K^+$ currents were evoked. The outward $K^+$ currents were markedly increased by the addition of 10 ${\mu}M$ sodium nitroprusside (SNP). 10 ${\mu}M$ S-nitroso-N-acetylpenicillamine (SNAP) and 1 mM 8-Bromo-cyclic GMP (8-Br-cGMP) also showed a similar effect to that of SNP. 1 mM tetraethylammonium (TEA) significantly reduced depolarization-activated outward $K^+$ currents. SNP-enhanced outward $K^+$ currents were blocked by the application of TEA. High EGTA containing pipette solution (10 mM) reduced the control currents and also inhibited the SNP-enhanced outward $K^+$ currents. 5 mM 4-aminopyridine (4-AP) significantly reduced the control currents but showed no effect on SNP-enhanced outward $K^+$ currents. 0.3 ${\mu}M$ apamin and 10 ${\mu}M$ glibenclamide showed no effect on SNP-enhanced outward $K^+$ currents. 10 ${\mu}M$ 1H-[1,2,4]oxadiazolo [4,3-a]quinoxaline-1-one (ODQ), a specific inhibitor of soluble guanylate cyclase, significantly blocked SNP-enhanced $K^+$ currents. We conclude that NO donors activate the $Ca^{2+}-activated$$K^+$ channels in guinea-pig ileal smooth muscle via activation of guanylate cyclase.
Kim, Chul-Soo;Jun, Jae-Yeoul;Kim, Sung-Joon;So, In-Suk;Kim, Ki-Whan
The Korean Journal of Physiology
/
v.29
no.2
/
pp.233-241
/
1995
The nonapeptide bradykinin has been shown to exhibit an array of biological activities including relaxation/contraction of various smooth muscles. In order to investigate the effects of bradykinin on the contractility and the electrical activity of antral circular muscle of guinea-pig stomach, the isometric contraction and membrane potential were recorded. Also, using standard patch clamp technique, the $Ca^{2+}-activated$ K currents were recorded to observe the change in cytosolic $Ca^{2+}$ concentration. $0.4 {\mu}M$ bradykinin induced a triphasic contractile response (transient contraction-transient relaxation-sustained contraction) and this response was unaffected by pretreatment with neural blockers (tetrodotoxin, atropine and guanethidine) or with apamin. Bradykinin induced hyperpolarization of resting membrane potential and enhanced the amplitude of slow waves and spike potentials. The enhancement of spike potentials was blocked by neural blockers. Both the bradykinin-induced contractions and changes in membrane potential were reversed by the selective $B_2$-receptor antagonist $(N{\alpha}-adamantaneacetyl-_{D}-Arg-[Hyp, Thy,_{D}-Phe]-bradykinin)$. In whole-cell patch clamp experiment, we held the membrane potential at -20 mV and spontaneous and transient changes of Ca-activated K currents were recorded. Bradykinin induced a large transient outward current, consistent with a calcium-releasing action of bradykinin front the intracellular calcium pool, because such change was blocked by pretreatment with caffeine. Bradykinin-induced contraction was also blocked by pretreatment with caffeine. From these results, it is suggested that bradykinin induces a calciumrelease and contraction through the $B_{2}$ receptor of guinea-pig gastric smooth muscle. Enhancement of slow wave activity is an indirect action of bradykinin through enteric nerve cells embedded in muscle strip.
The effects of $Cd^{2+}$ on spontaneous contraction, and the contractures induced by $0mM\;Na^+,\;60mM\;K^+\;and\;10^{-6}\;M$ acetylcholine, 1mM caffeine were studied in order to elucidate diverse actions of $Cd^{2+}$ on the $Ca^{2+}$ mobilization related with contractility in the antral circular muscle of guinea pig stomach. $Cd^{2+}$ inhibited the spontaneous contraction in a does dependent manner $(10^{-6}\;M\;10^{-4}\;M).\;Cd^{2+}\;(3{\times}10^{-5}M)$ suppressed 60 mM $K^+$ induced contracture composed or a phasic and a tonic response and the increased tonic response by the increased external $Ca^{2+}$ concentration. $Cd^{2+}$ also suppressed acetylcholine induced contracture composed of repetitive phasic and a tonic component and the increased tonic response by the increased external $Ca^{2+}$ concentration. Caffeine in the concentration of 1mM evoked contracture but $Cd^{2+}$ suppressed the contracture. $Cd^{2+}$ suppressed the amplitude of the $Na^+$ tee contracture dose dependently and the amplitude of $Na^+$ free contracture almost decreased to 20% of control amplitude in the concentration of $10^{-4}\;M\;Cd^{2+}$. From the above results, it is suggested that $Cd^{2+}$ may inhibit not only $Ca^{2+}$ influx via voltage sensitive, receptor operated $Ca^{2+}$ channel and Na/ca exchange but also intracellular $Ca^{2+}$ release from the sarcoplasmic reticulum in the antral circular muscle of guinea pig stomach.
$Mg^{2+}$ is an important regulator of many cardiac functions. However, regulation of intracellular $Mg^{2+}$ activity in the heart is not well characterized. To assess the effect of histamine $H_2$-receptor stimulation on intracellular $Mg^{2+}$ regulation, changes in extracellular $Mg^{2+}$ concentration were examined under a variety of conditions in perfused guinea pig hearts. $Mg^{2+}$ in the cardiac perfusate was measured by atomic absorbance spectrophotometry. The histamine ($10^{-6}$ M) inuced a marked $Mg^{2+}$ efflux from the heart. The $H_2$-receptor antagonists, cimetidine ($10^{-6}$ M), ranitidined ($10^{-5}$ M), but not a H1-receptor antagonist, diphenhydramine ($3{\times}10^{-6}$ M), completely blocked the histamine-induced $Mg^{2+}$ efflux. The $Mg^{2+}$ efflux could also be induced by forskolin ($3{\times}10^{-6}$ M), 8-Cl-cAMP ($2{\times}10^{-4}$ M), permeable cAMP analogue, or dimaprit, ($10^{-5}$ M). However, the carbachol ($10^{-5}$ M) considerably decreased the efflux of $Mg^{2+}$. In the presence of papaverine ($10^{-5}$ M), a phosphodiesterase inhibitor, dimaprit-induced $Mg^{2+}$ efflux was potentiated. These results suggest that a significant $Mg^{2+}$ efflux from perfused guinea pig heart by histamine can be induced by the histamine $H_2$-receptor stimulation and it is suggested that cytosolic cAMP may be linked.
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