• Natural Product Sciences
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• 제10권5호
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• pp.211-216
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• 2004

Citrus unshiu (Rutaceae) has long been known as an anti-inflammatory and anti-allergic agent. In the present study, the inhibitory effect of CUWE (Citus unshiu water extract) on the production of $TNF-{\alpha}$ and tryptase was examined. In addition, a possible mechanism for the inhibition of trypsin-stimulated human leukemic mast cell-1 (HMC- 1 ) activation was determined. To do so, $TNF-{\alpha}$ production from the HMC-1 cells that were stimulated by trypsin (100 nM) in the presence or absence of CUWE $(10,\;100,\;and\;100\;{\mu}g/ml)$ was measured by enzyme-linked immunosorbent assay (ELISA) and reverse transcription-PCR. The tryptase production was evaluated by reverse transcription-PCR. Extracellular signal-regulated kinase (ERK) activation was analyzed by Western blot. Trypsin activity was measured by using Bz-DL-Arg-p-nitroanilide (BAPNA) as substrate. Results showed that the CUWE inhibited production of both $TNF-{\alpha}$ and tryptase from the trypsin-stimulated HMC-1 in a dose-dependent manner. The CUWE a1so inhibited the ERK phosphorylation and trysin activity. These results indicate that the CUWE had an inhibitory effect on $TNF-{\alpha}$ and the tryptase productions by blocking the ERK phosphorylation and trypsin activity.

• Natural Product Sciences
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• 제11권4호
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• pp.207-212
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• 2005

Tetragonia tetragonoides (Aizoaceae) has been known as an anti-cancer agent. The activation of proteinase-activated receptor-2 (PAR-2) by trypsin appears to play a role in inflammation. In the present study, we examined the inhibitory effects of Tetragonia tetragonoides water extract (TTWE) on the production of tumor necrosis $factor-{\alpha}\;(TNF-{\alpha})$ and tryptase in trypsin-stimulated human leukemic mast cells (HMC-1) expressing PAR-2. HMC-1 cells were stimulated with trypsin in the presence or absence of TTWE (10, 100, and $1000\;{\mu}g/ml$). The level of $TNF-{\alpha}$ secretion from HMC-1 cells was measured by enzyme-linked immunosorbent assay (ELISA). $TNF-{\alpha}$ and tryptase mRNA expression were examined by reverse transcription-PCR. Also, extracellular signal-regulated kinese (ERK) activation was assessed by Western blot analysis. Trypsin activity was measured using the substrate Bz-DL-Arg-p-nitroanilide (BAPNA). It was observed that $TNF-{\alpha}$ secretion, tryptase mRNA and $TNF-{\alpha}$ mRNA expression in trypsin-stimulated HMC-1 cells were inhibited by pretreatment of TTWE ($1000\;{\mu}g/ml$). Furthermore, the pretreatment of TTWE ($1000\;{\mu}g/ml$) resulted in the reduction of ERK phosphorylation and trypsin activity. These results suggest hat TTWE might have the inhibitory effects on the PAR-2-dependent inflammation processes and it is likely to function as PAR-2 antagonist.

• 대한약학회:학술대회논문집
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• 대한약학회 2002년도 Proceedings of the Convention of the Pharmaceutical Society of Korea Vol.2
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• pp.254.2-254.2
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• 2002

Mast cells play an important role in inflammation by functioning as a source of histamine, tryptase, and proinflammatory cytokines. Lonicera Japonica (Caprifoliaceae. Lc) has been used to treat inflammation. We investigated whether the water extract of Lonicera Japonica(Lc) inhibit production of inflammatory mediators such as tryptase and tumor-necrosis factor (TNF)-${\alpha}$, and phosphorylation of extracellular signal-regulated kinase(ERK) in trypsin-stimulated HMC-1. (omitted)

• Asian Pacific Journal of Cancer Prevention
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• 제14권5호
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• pp.2955-2958
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• 2013

Background: Mast cells can influence tumor progression via different pathways and increased mast cell density has been demonstrated in oral squamous cell carcinoma (OSCC). It has been shown that the serum tryptase level is elevated with some malignant tumours and may thus be a useful parameter. However, there are no data available about OSCC. The main aim of this study was the evaluation of mast cell tryptase (MCT) level in OSCC patient serum. Materials and Methods: In this cross-sectional, analytic study, the circulating levels of MCT were assessed in sera of 55 OSCC patients and 34 healthy individuals with ELISA technique. Results: The serum MCT level in OSCC patients was 12-14 ng/ml, which was not significantly higher than the healthy control group. While the serum level of MCT was higher with larger tumours, there was no apparent correlation with clinico-pathological features such as patient age, gender, tumor location, stage, nodal status, distant metastasis, histological grade and smoking. Conclusions: Our findings showed that despite the results obtained from studies of other malignant tumors, serum level of MCT in OSCC patients could not be a credited as a reliable indicator of the presence or progression of tumours.

• Natural Product Sciences
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• 제13권2호
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• pp.169-173
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• 2007

Luteolin is a major flavonoid of Lonicera japonica and has anti-inflammatory effect. The activation of proteinase-activated receptor (PAR)-2 and -4 by trypsin appears to play a role in inflammation, In the present study, we examined the inhibitory effects of luteolin on activation of trypsin-induced human leukemic mast cells (HMC-1). HMC-1 cells were stimulated with trypsin, PAR-2 and PAR-4 agonist, in the presence or absence of luteolin. The level of TNF-${\alpha}$ secretion was measured by enzyme-linked immunosorbent assay (ELISA). The expression of tryptase and phosphorylated-extracellular signal-regulated kinase (ERK) were assessed by Westem blot analysis. Moreover, trypsin activity was measured by the substrate Bz-DL-Arg-p-nitroanilide (BAPNA). TNF-${\alpha}$ secretion and Tryptase expression in trypsin-stimulated HMC-1 cells were markedly inhibited by pretreatment of luteolin. Furthermore, the pretreatment of luteolin resulted in the reduction of ERK phosphorylation and trypsin activity. These results suggest that luteolin might has the inhibitory effects on the PAR-2 and -4-dependent inflammation.

• 고신대학교 의과대학 학술지
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• 제33권3호
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• pp.468-476
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• 2018

Perioperative anaphylaxis, although rare, is a severe, life-threatening unexpected systemic hypersensitivity reaction. Simultaneous administration of various drugs during anesthesia, the difficulty of communicate with patients in sedation and anesthesia, and coverage of the patient with surgical drapes are considered to be factors that impede early recognition of anaphylactic reactions. It is very important to perform an intradermal skin test because antibiotics are the most common cause of perioperative anaphylaxis. We report a case of negative-intradermal skin test antibiotic anaphylaxis mistaken for local aesthetic systemic toxicity without increase of serum tryptase for confirmative diagnostic biomaker during surgery under brachial plexus block. It is not possible to exclude the danger of anaphylaxis completely, even if it is negative-intradermal skin test and normal tryptase level. Therefore, anesthesiologists should be closely monitored and treated early for antibiotics related hypersensitive reaction, like other medicines during anesthesia.

• 미생물학회지
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• 제40권3호
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• pp.254-256
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• 2004

Polymerase chain reaction (PCR) is a powerful tool for precisely amplifying selected DNA sequences that have had a broad impact on genomic studies. When examining human $\alpha$- and $\beta$- tryptase genes which have 95% DNA homology, inconsistent PCR amplification of genomic sequences hampered our progress. This study suggests that long PCR technique on the original DNA digested with restriction enzymes improves both efficiency and sensitivity of PCR. These improved results seem to derived from the effective denaturation of the original genomic DNA template or reduction of formation of secondary structures that block either primer annealing or extension in PCR. Elimination of homo- or hetero-duplex products derived from highly homologous genes provides an additional advantage in this study. This communication describes how the use of restriction enzymes improved these efficiencies, and also facilitated studies of highly homologous genes including tryptase genes.

• Archives of Pharmacal Research
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• 제27권11호
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• pp.1141-1146
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• 2004

Lonicera japonica Thunb.(Caprifoliaceae) has long been known as an anti-inflammatory. In the present study, the effect of water fraction of Lonicera japonica (LJ) on trypsin-induced mast cell activation was examined. HMC-1 cells were stimulated with trypsin (100 nM) in the presence or absence of LJ (10, 100, and 1000 $\mu$ g/mL). TNF-$\alpha$ and tryptase production were measured by enzyme-linked immunosorbent assay (ELISA) and reverse transcription-PCR. Extracellular signal-regulated kinase (ERK) phosphorylation was assessed by Western blot. Trypsin activity was measured by using Bz-DL-Arg-p-nitroanilide (BAPNA) as substrate. LJ (10, 100, and 1000 $\mu$g/mL) inhibited TNF-$\alpha$ secretion in a dose-dependent manner. LJ (10, 100, and 1000 $\mu$g/mL) also inhibited TNF-$\alpha$ and tryptase mRNA expression in trypsin-stimulated HMC-1. Furthermore, LJ inhibited trypsin-induced ERK phosphorylation. However, LJ did not affect the trypsin activity even 1000 $\mu$g/mL. These results indicate that LJ may inhibit trypsin-induced mast cell activation through the inhibition of ERK phosphorylation than the inhibition of trypsin activity.

• 생명과학회지
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• 제19권12호
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• pp.1808-1814
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• 2009

방사선에 대한 방호와 면역기능 조절을 목적으로 새로운 생약복합조성물인 HemoHIM을 개발하였다. 식품 원료로 사용 가능한 생약재 3종 당귀, 천궁, 백작약의 에탄올 분획을 열수추출물에 첨가하여 HemoHIM을 제조하였다. HemoHIM의 항알레르기 효과를 검증하기 위하여 사람 비만세포주 HMC-1을 사용해 compound 48/80으로 유도되는 히스타민 분비량과 PMA/A23187로 유도되는 염증성 사이토카인의 분비량을 측정하였다. 히스타민의 양은 형광분석법으로, 염증성 사이토카인 IL-6, IL-8, TNF-$\alpha$, GM-CSF의 양은 효소결합 면역측정법으로 측정하였다. 저농도의 HemoHIM에 의해 히스타민 분비량이 억제되었고, 모든 농도에서 IL-6, TNF-$\alpha$, GM-CSF의 분비량은 억제되었지만 IL-8은 고농도에서만 억제되었다. 사이토카인의 mRNA 발현량은 HemoHIM의 농도 의존적으로 억제되었다. 그리고 c-kit와 Fc$\varepsilon$RI의 mRNA 발현량도 모든 농도에서 억제되었지만, tryptase의 mRNA 발현량은 저 농도에서만 억제되었다. 이상의 결과로 HemoHIM이 비만세포의 활성을 억제하는 효과가 있다는 것을 알 수 있었으며, 상대적으로 독성이 적은 항알레르기 제재로 개발할 가능성을 제시한 것으로 생각된다.

• 대한약학회:학술대회논문집
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• 대한약학회 2003년도 Proceedings of the Convention of the Pharmaceutical Society of Korea Vol.2-1
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• pp.92-92
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• 2003

The drug discovery landscape is changing rapidly in the post-genomic era. Mapping of the human genome has led to an abundance of potential drug targets. Drug discovery times and costs can be significantly reduced by developing methods for high throughput target identification/ validation, multiplexed assay development and high efficient combinatorial chemistry. (omitted)