Inhibitory Effect of Tetragonia tetragonoides Water Extract on the Production of $TNF-{\\alpha}$ and Tryptase in Trypsin-Stimulated Human Mast Cells원문보기
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 p...
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.
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.
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제안 방법
1984). Samples were added to each tube containing 2.85 mL of 0.1 M Tris-HCl (pH 8.0) and IM glycerol, and the assay was started with the addition of 20 mM substrate in DMSO. After 30 min incubation at 37°C, the reactions were stopped by the addition of 50 |丄 1 of glacial acetic acid, and the absorbance was measured at 405 nm with spectrophotometer and compared to those given by standard curves of p-nitroanilide to determine the concentration of product released.
대상 데이터
Plant materials and extract preparations - Herba of Tetragonia tetragonoides was purchased from an Oriental drug store, Daehak Hanyakkuk (Iksan, Korea) and identified by professor Dong-Yeul Kwon at the College of pharmacy, Wonkwang University. Voucher specimen (No.
데이터처리
for a number of experiments. Statistical significance was compared between each treated group and control by the Student's /-test. Each experiment was repeated at least three times and yielded comparable results.
성능/효과
, 1995). In this study, SBTI (10 μM) and TTWE (1000 pig/ml) showed 94% and 32% inhibition on the trypsin activity, respectively. Furthermore, SBTI and TTWE significantly inhibited trypsin-induced TNF-a and tryptase production in HMC-1 cells.
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