• Advances in Traditional Medicine
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• 제10권1호
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• pp.7-12
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• 2010

Typha orientalis' stem (TOS) is traditionally used as an herbal medicine for difficulty in urination, galactophoritis purulenta, whooping cough, and allergic dermatitis. However, its effect in experimental models remains unknown. Here, we report the effect of TOS on the phorbol 12-myristate 13-acetate (PMA) plus calcium ionophore A23187-induced inflammatory cytokine production and extracellular signal-regulated kinase (ERK) activation in the human mast cell line, HMC-1. TOS inhibited PMA plus A23187-induced cytokines such as tumor necrosis factor-alpha (TNF-$\alpha$) and interleukin (IL)-6. Maximal inhibition rate of TNF-$\alpha$ and IL-6 production by TOS (1 mg/ml) was about 44.02%, and 45.20%, respectively (P < 0.05). In addition, TOS inhibited the expression of TNF-$\alpha$ and IL-6 mRNA under the same condition. Moreover, TOS partially blocked PMA plus A23187-induced ERK phosphorylation. These results suggested TOS could inhibit the cytokine production through blocking of ERK activity.

• Journal of Life Science
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• 제11권1호
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• pp.7-10
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• 2001

Nonactin is the parent compound of a group of ionophore antibiotics, that known as the macrotetrolides. In previous report, in th course of screening superoxide radical-generating compounds from microbial sources, we first screened Streptomyces viridochromogenes JM-4151 that produces nonactin. It was proved that nonactin is superoxide radical-producing compound. In present study, we examined the optimal culture conditions of nonacin. Th optimal culture conditions for nonactin production were as follows: 1% soluble starch, 1% yeast extract, 0.2% ammonium nitrate, 0.06% magnesium sulfate, 0.2% calcium carbonate, initial pH 7.0 at 28$^{\circ}C$ for 96 h. The highest nonactin production was achieved in the production medium of initial pH7.0 at 28$^{\circ}C$ for 96h. The threshold level of dissolved oxygen was found to be above 33.2% at 28$^{\circ}C$ when 1% soluble starch was used as a carbon source. These results suggest that S. viridochromogenes JM-4151 might be a possible strain for industrial nonactin producer.

• 한국응용약물학회:학술대회논문집
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• 한국응용약물학회 1994년도 춘계학술대회 and 제3회 신약개발 연구발표회
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• pp.274-274
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• 1994

본 연구에서는 토끼신장 근위세뇨관 일차배양세포에서 브라디키닌의 생리작용이 phospholipase D (PLD)에 의해 매개되는지를 살펴 보기위해 PLD 효소반응의 특이한 성질인 transphosphatidylation 반응의 생성물인 phosphatidylethanol (PEth) 의 세포내 양을 측정함으로 PLD 효소의 관련성을 규명할 수 있었다. 시간경과에 따른 phosphatidic acid (PA) 및 diacylglycerol (DAG) 의 생성을 살펴본 결과 PA가 DAG보다 먼저 생성되어 최고치 (30초)에 도달하였고 DAG는 1분이후부터 5분까지 서서히 생성되는 양상을 나타내었다. 또한 0.5에서 5％까지의 에탄올 존재하에 PA 및 PE소 생성량을 비교해본 결과 에탄올량이 증가함에 따라 PA는 감소하는 반민 PEth 의 생성은 계속 증가하였다. 한편 브라디키닌 농도 변화 실험에서는 브라디키닌농도가 증가함에 따라 PA 및 PEth 둘다 생성이 증가되었다. 이러한 결과로부터 토끼신장 근위세뇨관 세포막에 존재하는 브라디키닌수용체는 브라디키닌에 의해 activation 시 PLD를 직접적으로 활성화시켜 그들의 작용을 세포내로 전달한다는 사실을 알 수 있었다. 또한 PLD 효소활성의 activator로 수용체효능 제외에 칼슘이온, protein kinase C (PKC) 등이 몇몇 다른 실험에 의해 밝혀져 있고, G protein 역시 PLD 효소 활성을 조절하는 역할이 있음이 알려졌다. calcium ionophore 및 칼슘채널길항제인 verapamil을 이용한 실험에서 우리는 브라디키닌의 PLD 활성화는 칼슘이온에 의존적인 경로 및 비의존적인 경로가 같이 존재함을 알수 있었다. 또한 브라디키닌의 PLD 활성화기전이 PKC 의존적인지를 살펴보기위해 PKC activator(PMA) 및 inhibitor (staurosporine)를 이용한 실험에서 브라디키닌은 신장세포에서 PKC를 통하여 PLD를 활성화시킴으로 신호전달을 하는 것으로 추측되었다. 마지막으로 가수분해안되는 G protein 유도체인 GTPrS 및 G protein 활성물질 NaF, 백일해독소등을 이용한 실험에서 G protein 의 PLD 조절활성을 확인할 수 있었다.

• 대한한방내과학회지
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• 제25권2호
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• pp.174-182
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• 2004

Object : This study investigates Jaeumganghwa-tang(JGT) has been used for the purpose of prevention and treatment of allergic inflammatory diseases. This study was to investigate the biological effects of JGT. Methods : Cytotoxcicity and inflammatory cytokines secretion with human mast cells(HMC-1) were examined. HMC-1 cells were stimulated with phorbol l2-myristate 13-acetate (PMA) and calcium ionophore A23l87. JGT by itself had no effect on cytotoxicity of HMC-1. The effects of JGT on the secretion of tumor necrosis factor-alpha(TNF-${\alpha}$) and interleukin(IL)-6 from HMC-1 were evaluated with enzyme-linked immunosorbent assay(ELISA). Result : It was found that JGT inhibited PMA plus A23187-induced TNF-${\alpha}$ and IL-6 secretion. JGT also inhibited the $NF-{\kappa}$B(p50) expression. Conclusion : These results suggest that JGT inhibits the secretion of inflammatory cytokines in HMC-1 cells through blockade of $NF-{\kappa}B$ activation. Taken together, these effects support a role for JGT as a therapeutic agent in treatment of allergic inflammatory diseases such as asthma.

• 대한한의학방제학회지
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• 제25권2호
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• pp.167-177
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• 2017

Objectives : This study aimed to evaluate inhibitory effects of GHJ on allergic inflammatory response in human mast cells (HMC-1). Methods : To investigate the inhibitory effect of GHJ (62.5, 125, 250, 500, $1000{\mu}g/mL$), HMC-1 cells were stimulated with phorbol 12-myristate 13-acetate plus calcium ionophore A23187 (PMACI). Enzyme-linked immunosorbent assays (ELISAs), RT-PCR and Western blot analysis were investigated using GHJ extract. Results : GHJ inhibited levels of $TNF-{\alpha}$ and IL-6 of $1000{\mu}g/mL$ concentration in ELISA and mRNA expression. GHJ had inhibitory effects in level of MAPKs, $p-I{\kappa}B-{\alpha}$ and p-NF-kB also. GHJ attenuated Compound 48/80-stimulated histamine release. In addition, GHJ inhibited PCA reaction in vivo. Conclusion : This study indicated that GHJ extract can inhibit allergic responses in HMC-1 cell.

• 한국동물번식학회:학술대회논문집
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• 한국동물번식학회 2001년도 춘계학술발표대회
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• pp.66-66
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• 2001

Since the first birth of pig derived from embryonic cells by nuclear transfer, many researches to produce cloned pig have been carried out. Recently, two reports about the birth of somatic cell cloned pigs using in vivo oocytes and also Betthauser et al. (2000) reported the birth of somatic cell cloned pigs using in vitro oocytes. So here we investigated the effect of activation method and culture medium on in vitro development of porcine nuclear transfer embryo using fetal fibroblast. Oocytes derived from slaughter house obtained ovaries were matured for 42 to 44 h in TCM 199. Matured oocytes were denuded using 0.1% hyaluronidase and then Oocytes with the first polar body were used for enucleation by aspirating the first polar body and adjacent cytoplasm in TCM 199 supplemented with 7.5 $\mu\textrm{g}$ cytochalasin B. Petal fibroblast cells were prepared from 35 days old fetus. To be used as donor cells, fetal fibroblast cells were serum starved for 3 to 5 days and then isolated into single co:1 by trypsinization. Nuclear transfer embryos were fused using 2 times 1.25㎸ for 30$mutextrm{s}$. Fused NT embryos were activated with calcium ionophore (CI) and 6-dimethyl-aminopurine (6-DMAP). Activated oocytes were cultured in NCSU 23 or BECM 3 for 6 days. There was no significant difference between chemical activation and no chemical activation for blastocyst development rate(11.6 vs. 14.8%). However, cell number was significantly higher when NT embryos were activated with CI and 6-DMAP (31.2 vs. 22.6). When NT embryos were cultured in NCSU 23 or BECM 3, blastocyst development rate was 16.4 and 13.2%, respectively, and cell number was 31.5 and 24.1, respectively. These results suggest that chemical activation after fusion and culture in NCSU 23 could increase cell number of porcine NT embryos.

• Reproductive and Developmental Biology
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• 제32권2호
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• pp.97-104
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• 2008

Plasminogen activators (PAs) are serine protease that cleave plasminogen to form the active protease plasmin. PA/plasmin system playa role in mammalian fertilization and motility and acrosome reaction of sperm. The present study was undertaken to identify PAs in porcine gametes and investigate a possible role of plasminogen in in vitro fertilization in the pig. When boar spermatozoa were preincubated in a fertilization medium (mTBM) for 0, 2, 4 or 6 h, the activity of tPA-PAI ($110{\sim}117\;kDa$), tPA ($62{\sim}70\;kDa$), and uPA ($34{\sim}38\;kDa$) was observed in the sperm incubation medium and sperm sample. PA activities in the sperm incubation medium significantly (p<0.05) increased according to increasing incubation times, while PA activities in sperm significantly (p<0.05) decreased at the same times. In addition, the rate of acrosome reaction in spermatozoa increased by increasing culture times. When oocytes were separated from porcine cumulus-oocytes complexes at 0, 22 or 44 h of maturation culture, no PA activities were observed in cumulus free-oocyte just after aspiration from follicles. However, the activity of tPA-PAI ($108{\sim}113\;kDa$) and tPA ($75{\sim}83\;kDa$) was observed at 22 h of in vitro culture and significantly (p<0.05) increased as the duration of the culture increased. On the other hand, when porcine oocytes were activated by sperm penetration or calcium ionophore, plasminogen significantly (p<0.05) increased ZP dissolution time (sec) in activated oocytes by sperm penetration. These results suggest that supplementation of plasminogen to fertilization medium may playa positive role in the improvement of in vitro fertilization ability in the pig.

• 한방안이비인후피부과학회지
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• 제22권2호
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• pp.60-73
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• 2009

Objective : Houttuyniae Herba is widely used in oriental medicine as a remedy for inflammation. However, as yet there is no clear explanation of how Houttuyniae Herba affects the production of inflammatory cytokine. This study was to determine the effects of Essence extracted from Houttuynia cordata Thunb(HCT) on the mast cell-mediated inflammatory responses. Method : We measured the amount of inflammatory cytokine production induced by the phorbol myristate acetate (PMA) plus calcium ionophore(A23187) in the human mast cell line (HMC-1) incubated with various concentrations of HCT. The TNF-$\alpha$, IL-6 and IL-8 secreted protein levels were measured by the ELISA assay. The TNF-$\alpha$, IL-6 and IL-8 mRNA levels were measured by the RT-PCR analysis. Nuclear and cytoplasmic proteins were exmined by Western blot analysis. The NF-$\kappa$B promoter activity was examined by a luciferase assay. Result : HCT inhibited the PMA + A23187-induced TNF-$\alpha$, IL-6 expression and reduced mRNA of TNF-$\alpha$, IL-6 and IL-8. we observed that HCT suppressed the induction of NF-B activity. In addition, HCT suppressed PMA plus A23187-induced NF-$\kappa$B promoting activity. Conclusion : In this study, we have found that HCT is an inhibitor of NF-$\kappa$B and cytokines on the mast cell-mediated inflammatory responses.

• 한국자원식물학회:학술대회논문집
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• 한국자원식물학회 2018년도 추계학술대회
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• pp.128-128
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• 2018

Nypa fruticans wurmb (NF) has been used as traditional medicinal food in Asian countries. Especially, NF has been used for conventional medicine to treat inflammatory periodontal diseases. Previous studies have been shown that NF has large amount of useful constituents such as phenolic acids, polyphenols and flavonoids. Also, NF is known as having medicinal effects such as anti-oxidant, anti-inflammatory and cholesterol-lowering effects. NF has recently been attracted to use complementary medicinal food on inflammatory diseases in Korea. However, there are no obvious effects in inflammatory and metabolic diseases also mechanisms has been studied yet. The purpose of this study was to investigate the anti-inflammatory effects of NF and steamed-NF (SNF), which recently has been used as health food, using Human keratinocyte cell line (HaCaT) and Human mast cell line (HMC-1). The cytotoxicities of NF and SNF were measured by using MTT assays in HaCaT cells and HMC-1 cell. To evaluate anti-inflammatory effects of NF and SNF, HaCaT cells were stimulated with tumor necrosis factor $(TNF)-{\alpha}$ and Interferon $(IFN)-{\gamma}$. Also, HMC-1 cells were stimulated with phorbol-12-myristate-13-acetate (PMA) and A23187 calcium ionophore (A23187) to induce allergic inflammation. Inflammatory cytokine were measured by enzyme-linked immunosorbent assay (ELISA). In this result, the extract of NF and SNF (0.01 - 1mg/ml) did not show cytotoxicity in HaCaT cells and HMC-1 cells. In addition, the NF and SNF suppressed the production of interleukin (IL)-6 and IL-8 in HaCaT cells at highest concentration. Furthermore, the treatment of SNF significantly inhibited the secretion level of IL-8 in PMA plus A23187-stimulated HMC-1 cells compared with NF treatment group. These results suggest that the extract of NF and SNF may serve as a potential therapy for skin inflammatory diseases.

• 한국약용작물학회지
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• 제15권2호
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• pp.132-137
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• 2007

Inhibitory effect of Scutellaria baicalensis ethanol extracts (SR) on chemical mediator release and immunoglobulin (Ig) production from Sprague-Dawley rats originated cells as type I allergic reaction was examined. SR showed concentration-dependent inhibition on basal and concanavalin A (ConA)-stimulated Ig production. In the mesenteric lymph node lymphocytes, the inhibitory effect of SR on the IgE production in the presence of Con A was stronger than these on IgA and IgG production. Moreover, tumor necrosis factor-alpha $(TNF-{\alpha})$ production-inhibiting effect of SR in the presence ConA was observed. However, SR did not affect the production of $interferon-{\gamma}$. SR also inhibited histamine release from the peritoneal exudate cells stimulated with a calcium ionophore A23187. In the case of leukotriene B4, SR markedly inhibited it at the concentration of 100 mg/ml. From these results, ethanol extracts obtained from Scutellaria baicalensis may have an anti-allergic effect on the intestinal system of rats.