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In vitro and in vivo anti-inflammatory and anti-allergic responses caused by water extract of Mori Cortex (상백피(桑白皮)가 항알러지 및 항염증반응에 미치는 영향)

  • Lee, Jin-Yong;Kim, Deog-Gon;Cho, Hyung-Jun
    • The Journal of Pediatrics of Korean Medicine
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    • v.19 no.2
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    • pp.175-195
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    • 2005
  • Objective: This experimental study was performed to examine the in vitro and in viva anti-inflammatory and anti-allergic effects of Mori Cortex. Methods: Water extract of Mori Cortex was studied to its ability to stimulate or inhibit macrophage 264.7 cells to produce inflammatory and allergic mediators. Cytokines such as $IL-1{\beta}$, IL-6, IL-10 and $TNF-{\alpha}$ were measured by immunochemical assay. In vitro, the macrophages 264.7 were classified into four groups. One group was a normal group. The other group was a (-) control group stimulated with LPS. And the third group was a (+) control group pretreated for 1 hour with hydrocortisone. And the fourth group was a sample group pretreated for 1 hour with Mori Cortex. After pretreatment, macrophage were incubated with lipopolysaccharide(LPS) $100\;ng/m{\ell}$ for 12 hour and media collected and $IL-1{\beta}$, IL-6, IL-10 and $TNF-{\alpha}$ concentrations in supernatants were measured each by Enzyme linked immuno-soubent assay. Mori Cortex were used $50\;{\mu}g/m{\ell},\;100\;{\mu}g/m{\ell},\;250\;{\mu}g/m{\ell},\;500\;{\mu}g/m{\ell},\;and\;1,000\;{\mu}g/m{\ell}$. Hydrocortisones were used $10^{-8}M,\;10^{-7}M,\;10^{-6}M,\;10^{-5}M\;and\;10^{-4}M$. In vivo, the SD rats were classified into three groups. One group was a normal group injected with normal saline into the abdominal cavity. The other was a control group prescribed to compound 48/80 after normal saline injection. And the third was a sample group prescribed to compound 40/80 after Mori Cortex injection. Then, the release of histamine, IL-6 and $TNF-{\alpha}$ were measured. Results : In vitro, Man Cortex significantly increased the release of $IL-1{\beta}\;and\;TNF-{\alpha}$ by LPS-stimulated macrophage 264.7 cells. And it significantly decreased the release of IL-10. In IL-6, Mori Cortex of low concentration significantly decreased the release of IL-6, but that of high concentration acted in reverse. In vivo, Man Cortex didn't show significant inhibitory effects on the release of histamine and IL-6 in comparison with that of the control group. But it significantly increased the release of $TNF-{\alpha}$ in comparison with that of the control group.

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Effects of Gagam-jeonggitang, Gami-hwajeongjeon and Gami-tonggyutang on secretion of airway mucus In Vitro and In Vivo (가감정기탕(加減正氣湯), 가미화정전(加味和正煎), 가미통규탕(加味通竅湯)이 기도점액 분비에 미치는 영향)

  • Han, Jae-Kyung;Kim, Yun-Hee;Chae, Ho-Youn
    • The Journal of Pediatrics of Korean Medicine
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    • v.21 no.1
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    • pp.117-137
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    • 2007
  • Objectives : In the present study, the author intended to investigate Gagam-jeonggitang(GJT), Gami-hwajeongjeon(GHJ) and Gami-tonggyutang(GTT) significantly affect in vivo and in vitro mucin secretion from airway epithelial cells. Methods : In vivo experiment, the author induced hypersecretion of airway mucin, hyperplasia of tracheal goblet cells and the increase in intraepithelial mucosubstances by exposing rats to SO2 during 3 weeks. Effects of orally-administered GJT, GHJ and GTT during 1 week on in vivo mucin secretion and hyperplasia of tracheal goblet cells were assesed using ELISA and staining goblet cells with alcian blue. For in vitro experiment, confluent HTSE cells were metabolically radiolabeled with 3H-glucosamine for 24 hrs and chased for 30 min in the presence of each agent to assess the effects of each agent on 3H-mucin secretion. Possible cytotoxicities of each agent were assessed by measuring lactate dehydrogenase release. Also, the effects of each agent on contractility of isolated tracheal smooth muscle and effects of each agent on MUC5AC gene expression in cultured HTSE cells were investigated. Results : GJT, GHJ and GTI inhibited hypersecretion of in vivo mucin: GJT and GHJ inhibited the increase of number of goblet cells. However, GTT did not affect the increase of number of goblet cells; GJT and GTT significantly increased mucin secretion from cultured HTSE cells, without significant cytotoxicity. GHJ increased mucin secretion and showed mild cytotoxicity at the highest concentration: GJT, GHJ and GTT chiefly affected the 'mucin' secretion; GJT, GHJ and GTT did not affect Ach-induced contraction of isolated tracheal smooth muscle; GTT did not significantly affect the expression levels of MUC5AC gene. However, GJT significantly. inhibit the expression levels of MUC5AC gene and GHJ significantly increased the expression levels of MUC5AC gene. These results suggest that GJT, GHJ and GTI can increase mucin secretion during short-term treatment(in vitro), whereas it can inihibit hypersecretion of mucin during long-term treatment(in vivo) and GJT and GHJ can not only affect the secretion of mucin but also affect the expression of mucin gene. Conclusions : The author suggests that the effects GJT, GHJ and GTT with their components should be further investigated and it is valuable to find, from oriental medical prescriptions, novel agents which might regulate hypersecretion of mucin from airway epithelial cells.

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Capsaicin Induces Acute Spinal Analgesia and Changes in the Spinal Norepinephrine Level (Capsaicin에 의한 척수 수준에서의 급성 진통효과와 Norepinephrine의 변화)

  • Park, Hyoung-SuP;Park, Kyung-Pyo
    • The Korean Journal of Pharmacology
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    • v.29 no.1
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    • pp.33-41
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    • 1993
  • Central analgesic effect of capsaicin was assessed by the tail flick reflex (TFR) test, using male Sprague-Dawley rats under anesthesia with pentobarbital sodium (induction with 40 mg/kg and maintenance with $4{\sim}8\;mg/kg/hr$). Level of norepinephrine in the spinal cord was also measured. Capsaicin, $35{\sim}150\;{\mu}g$, was injected intrathecally, and the TFR latency was measured before, 10, 30, and 60 minutes after the drug administration. TFR latency was increased 100% or more immediately by intrathecal capsaicin, from 2.9 seconds to the maximum of 7.0 seconds at 10 minute after the drug; P<0.01. The increase in TFR latency was maintained during the course of experiment of 2 hours. Concomitant reduction of NE content in the spinal cord was observed; from 16 ng/mg protein to 7 ng/mg protein. On the other hand, subcutaneous injection of capsaicin of 50 mg/kg did not change the TFR latency although the NE content reduced similarly to the case of intrathecal injection. Pretreatment of the animal with 0.5 mg/kg of MK-801 reversed the increase of TFR latency and NE reduction induced by intrathecal capsaicin. These results suggest that capsaicin causes analgesia at the spinal cord level by activating the excitatory amino acid-NE-dorsal horn interneurons axis of the descending inhibitory pain modulation pathway.

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Protective Effect of Phosphatidylcholine on Lipopolysaccharide-Induced Acute Inflammation in Multiple Organ Injury

  • Jung, Yoon Yang;Nam, Yunsung;Park, Yong Seol;Lee, Ho Sung;Hong, Soon Auck;Kim, Beom Keun;Park, Eon Sub;Chung, Yoon Hee;Jeong, Ji Hoon
    • The Korean Journal of Physiology and Pharmacology
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    • v.17 no.3
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    • pp.209-216
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    • 2013
  • Soybean polyunsaturated phosphatidylcholine (PC) is thought to exert anti-inflammatory activities and has potent effects in attenuating acute renal failure and liver dysfunction. The aim of this study was to investigate the effects of PC in protecting multiple organ injury (MOI) from lipopolysaccharide (LPS). Six groups of rats (N=8) were used in this study. Three groups acted as controls and received only saline, hydrocortisone (HC, 6 mg/kg, i.v.) or PC (600 mg/kg, i.p.) without LPS (15 mg/kg, i.p.) injections. Other 3 groups, as the test groups, were administered saline, HC or PC in the presence of LPS. Six hours after the LPS injection, blood and organs (lung, liver and kidney) were collected from each group to measure inflammatory cytokines and perform histopathology and myeloperoxidase (MPO) assessment. Serum cytokines (TNF-${\alpha}$, IL-6 and IL-10) and MPO activities were significantly increased, and significant histopathological changes in the organs were observed by LPS challenge. These findings were significantly attenuated by PC or HC. The treatment with PC or HC resulted in a significant attenuation on the increase in serum levels of TNF-${\alpha}$ and IL-6, pro-inflammatory cytokines, while neither PC nor HC significantly attenuated serum levels of IL-10, anti-inflammatory cytokine. In the organs, the enhanced infiltration of neutrophils and expression of ED2 positive macrophage were attenuated by PC or HC. Inductions of MPO activity were also significantly attenuated by PC or HC. From the findings, we suggest that PC may be a functional material for its use as an anti-inflammatory agent.

Developmental changes in GABAA tonic inhibition are compromised by multiple mechanisms in preadolescent dentate gyrus granule cells

  • Pandit, Sudip;Lee, Gyu Seung;Park, Jin Bong
    • The Korean Journal of Physiology and Pharmacology
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    • v.21 no.6
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    • pp.695-702
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    • 2017
  • The sustained tonic currents ($I_{tonic}$) generated by ${\gamma}$-aminobutyric acid A receptors ($GABA_{A}Rs$) are implicated in diverse age-dependent brain functions. While various mechanisms regulating $I_{tonic}$ in the hippocampus are known, their combined role in $I_{tonic}$ regulation is not well understood in different age groups. In this study, we demonstrated that a developmental increase in GABA transporter (GAT) expression, combined with gradual decrease in $GABA_AR{\alpha}_5$ subunit, resulted in various $I_{tonic}$ in the dentate gyrus granule cells (DGGCs) of preadolescent rats. Both GAT-1 and GAT-3 expression gradually increased at infantile ($P_{6-8}$ and $P_{13-15}$) and juvenile ($P_{20-22}$ and $P_{27-29}$) stages, with stabilization observed thereafter in adolescents ($P_{34-36}$) and young adults ($P_{41-43}$). $I_{tonic}$ facilitation of a selective GAT-1 blocker (NO-711) was significantly less at $P_{6-8}$ than after $P_{13-15}$. The facilitation of $I_{tonic}$ by SNAP-5114, a GAT-3 inhibitor, was negligible in the absence of exogenous GABA at all tested ages. In contrast, $I_{tonic}$ in the presence of a nonselective GAT blocker (nipecotic acid, NPA) gradually decreased with age during the preadolescent period, which was mimicked by $I_{tonic}$ changes in the presence of exogenous GABA. $I_{tonic}$ sensitivity to L-655,708, a $GABA_AR{\alpha}_5$ subunit inverse agonist, gradually decreased during the preadolescent period in the presence of NPA or exogenous GABA. Finally, Western blot analysis showed that the expression of the $GABA_AR{\alpha}_5$ subunit in the dentate gyrus gradually decreased with age. Collectively, our results suggested that the $I_{tonic}$ regulation of altered GATs is under the final tune of $GABA_AR{\alpha}_5$ subunit activation in DGGCs at different ages.

Partial Characterization of Physicochemical and Kinetic Properties of $Ca^{++}-ATPase$ System in Luteal Membranes (황체막에서의 $Ca^{++}-ATPase$의 특성)

  • Choi, Gyu-Bog;Koo, Bon-Sook;Kim, In-Kyo
    • The Korean Journal of Physiology
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    • v.20 no.2
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    • pp.257-270
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    • 1986
  • It has been reported that the luteal function may be regulated by the intracellular calcium in luteal cells (Higuchi et al, 1976; Dorflinger et at, 1984; Gore and Behrman, 1984) which is adjusted partially by $Ca^{++}-ATPase$ activities in luteal cell membranes (Verma and Pennistion, 1981). However, the physicochemical and kinetic properties of $Ca^{++}-ATPase$ in luteal membranes were not fully characterized. This study was, therefore, undertaken to partially characterize the physicochemical and kinetic properties of $Ca^{++}-ATPase$ system in luteal membranes and microsomal fractions, known as an one of the major $Ca^{++}$ storge sites (Moore and Pastan, 1978), from the highly luteinized ovary Highly luteinized ovaries were obtained from PMSG-hCG injected immautre female rats. Light membrane and heavy membrane fractions and microsomal fractions were prepared by the differential and discontinuous sucrose density gradient centrifugation method desribed by Bramley and Ryan (1980). Light membrane and heavy membrane fractions and microsomal fractions from highly luteinized ovaries are composed of the two different kinds of $Ca^{++}-ATPase$ system. One is the high affinity $Ca^{++}-ATPase$ which is activated in low $Ca^{++}$ concentration (Km, 10-30 nM), the other is low affinity $Ca^{++}-ATPase$ activated in higher $Ca^{++}$ concentration $(K_{1/2},\;40\;{\mu}M)$. At certain $Ca^{++}$ concentrations, activities of high and low affinity $Ca^{++}-ATPase$ are the highest in light membrane fractions and are the lowest in microsomal fractions. It appeares that high affinity $Ca^{++}-ATPase$ system have 2 binding sites for ATP (Hill's coefficient; around 2 in all membrane fractions measured) and the positive cooperativity of ATP bindings obviously existed in each membrane fractions. The optimum pH for high affinity $Ca^{++}-ATPase$ activation is around S in all membrane fractions measured. The lipid phase transition temperature measured by Arrhenius plots of high affinity $Ca^{++}-ATPase$ activity is around $25^{\circ}C$. The activation energies of high affinity $Ca^{++}-ATPase$ below the transition temperature are similar in each membrane fractions, but at the above transition temperature, it is the hightest in heavy membrane fractions and the lowest in microsomal fractions. According to the above results, it is suggested that intracellular $Ca^{++}$ level, which may regulate the luteal function, may be adjusted primarily by the high affinity $Ca^{++}-ATPase$ system activated in intracellular $Ca^{++}$ concentration range $(below\;0.1\;{\mu}M)$.

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The alterations of $Ca^{2+}$-activated $K^+$ channels in coronary artery during cardiac hypertrophy

  • Kim, Nari;Lee, Sang-Kyeong;Chung, Joon-Yong;Seog, Dae-Hyun;Kim, Euiyong;Jin Han
    • Proceedings of the Korean Biophysical Society Conference
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    • 2003.06a
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    • pp.23-23
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    • 2003
  • It has been suggested that the impairment of smooth muscle cell (SMC) function by alterations in the $Ca^{2+}$-activated $K^{+}$ ( $K_{Ca}$ ) channels accounts for the reduction in coronary reserve during left ventricular hypertrophy (LVH). However, this hypothesis has not been fully investigated. The main goal of this study was to assess whether the properties of $K_{Ca}$ channels in coronary SMCs were altered during LVH. New Zealand white rabbits (0.8-1.0 kg) and Sprague-Dawley rats (300-400 g) were randomly selected to receive either an injection of isoproterenol (300 $\mu\textrm{g}$/kg body weight) or an equal volume of 0.9% saline (1 mL/kg body weight). The animals developed LVH 10 days after injection. In patch-clamp experiments, the unitary current amplitude and open probability for the $K_{Ca}$ channels were significantly reduced in LVH patches compared with control patches. The concentration-response curve of the $K_{Ca}$ channel to [C $a^{2+}$]$_{i}$ was shifted to the right. Inhibition of the $K_{Ca}$ channels with TEA was more pronounced in LVH cells than in the control cells. The whole-cell currents of $K_{Ca}$ channels were reduced during LVH. Western blot analysis indicated no differences in $K_{Ca}$ channel expression between the control and LVH coronary SM membranes. In contraction experiments, the effect of a high $K^{+}$concentration on the resting tension of the LVH coronary artery was greater than on that of the control. The effect of TEA on the resting tension of the LVH coronary artery was reduced as compared with the effect on the control. Our findings imply a novel mechanism for reduced coronary reserve during LVH.ing LVH.

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Effect of Plant Extracts on the Activity of Alcohol Dehydrogenase and the Antioxidation in Alcohol-treated Rat Hepatocyte (알코올 투여한 흰쥐 간세포내 알코올 탈수소효소의 활성과 항산화에 미치는 식물추출물들의 영향)

  • 조성환;김지철;김성완
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.30 no.4
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    • pp.679-683
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    • 2001
  • This study was purposed to compare the activity of alcohol dehydrogenase and antioxidative effects of several plant extracts in the alcohol-treated rat liver. Sprague-Dawley rat weighing about 200 g were divided into the following 6 groups : normal, alcohol group and 4 different plant extracts administrated groups(Soybean sprout, Pine needle, Lentinus edodes, acanthopanacis cortex). Each plant extract was administrated orally by 200mg/kg b.w./day for 8 days before the alcohol treatment (5 g of 30% alcohol /kg b.w. by i.p.injection). All rats were sacrificed at 90 min after the alcohol treatment. The alcohol concentrations in serum of Soybean sprout and pine needle group were significantly lower than the Lentinus edodes and Acanthopanacis cortex group. The activity of alcohol dehydrogenase in the hepatic cytosol of Soybean sprout and Pine needle group was also significantly higher than the alcohol and the other groups However, the activity of catalase seemed not to be affected, although the extract groups showed slightly higher activities of catalase than the alcohol group. These results may indicate that the extracts of Soybean sprout and Pine needle were relatvely effective on the alcohol degradation. the activity of blutathione-peroxidase and lipid peroxidaton of all of the extract groups were significantly lower than the activity of alcohol group. These results can suggest that all of the use plant extracts more or less have an antioxidative effect on the alcohol-induced oxidation and especially, extracts of Soybean sprout and Pine needle have an stimulating effect on the alcohol absorption and degradation.

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Effect of Black Pepper on the Composition of Serum and Liver in the Rat (호초(胡椒)가 백서(白鼠)의 혈청(血淸) 및 간장성분(肝臟成分)에 미치는 영향(影響))

  • Cho, Soo Yeul;Lee, Sook Hwa
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.12 no.3
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    • pp.219-224
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    • 1983
  • This experiment was undertaken to investigate the effect of black pepper (BP) added to food on a living body and to determine the growth rate, digestibility, weight of organs, composition of blood and liver in the rat. Fifty-six male rats of Sprague-Dawley strain, weight $120{\pm}10g$, were divided into 4 groups and were fed ad libitum for 8 weeks. Experimental diets were divided into 4 groups. Each groups were separately added 0%, 0.5%, 2.0% and 5.0% of BP level. The results obtained are summarized as follows. The highest net weight of liver and heart generally increased according to increasing amount of BP and that of spleen and lung were not significantly different. Kidney weight was significantly higher in the group of containing 5.0% BP. Serum GOT and GPT were not significant, serum glucose was significantly lower in the group of containing 5.0% BP, and serum cholesterol was significantly higher in the group of containing 5.0% BP. Total serum protein decreased gradually as the amount of BP increased, but albumin and globulin were nor affect. Serum Na and K were not significant, but serum Ca and P were decreased as the amount of BP was increased. Liver crude lipid and crude protein were not affect. In fatty acid composition, arachidonic acid was significantly lower in the group of containing 5.0% BP.

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Hypoglycemic Effects of Basidiomycetes Mycelia and Cereals Fermented with Basidiomycetes (버섯 균사체 및 배양곡물의 혈당강하 효과)

  • Choi, Hee-Don;Seog, Ho-Moon;Park, Yong-Kon;Park, Young-Do;Kim, Jung-Ae
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.36 no.10
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    • pp.1257-1262
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    • 2007
  • To develop basidiomycetes-fermented cereals with hypoglycemic property, inhibitory effects of basidiomycetes mycelia and basidiomycetes-fermented cereals on postprandial glucose were investigated. In vitro effect of basidiomycetes mycelia on retarding the membrane transport of glucose was compared with pectin. For basidiomycetes mycelia, $13.1{\pm}3.6{\sim}41.8{\pm}8.0%$ of total glucose remained in inner solution of dialysis membrane after dialysis for 120 min, indicating that most of basidiomycetes mycelia might effectively retard membrane transport of glucose. Glucose tolerance of basidiomycetes mycelia and basidiomycetes-fermented cereals was tested on streptozotocin-induced diabetic rats administrated with maltose. Postprandial glucose levels of basidiomycetes mycelia, $389.4{\pm}43.8{\sim}426.3{\pm}49.4mg/dL$, were considerably lower than that of control, $535.3{\pm}78.6mg/dL$, at 30 minutes after maltose administration. Namely, basidiomycetes mycelia showed better postprandial glucose lowering effect than pectin. Brown rice and barley fermented with Paecilomyces japonica showed much lower postprandial glucose level than raw brown rice and barley, especially hypoglycemic effect of barley fermented with Paecilomyces japonica was significant.