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红球姜的粗乙醇提取物通过抑制人呼吸道上皮细胞中的ERK,抑制佛波酯诱导的MUC2和MUC5AC表达。

Crude ethanolic extracts of Zingiber cassumunar ROXB. inhibit PMA-induced MUC2 and MUC5AC expression via ERK inhibition in human airway epithelial cells.

作者信息

Limvuttegrijerat Thidarat, Poachanukoon Orapan, Koontongkaew Sittichai, Na Ayudhya Thaweephol Dechatiwongse

机构信息

Faculty of Medicine, Thammasat University, Pathum Thani 12120, Thailand.

出版信息

Asian Pac J Allergy Immunol. 2014 Dec;32(4):328-36. doi: 10.12932/AP0517.32.4.2014.

Abstract

BACKGROUND

Over expression of mucin often leads to serious airway pathologies. The rhizome of Zingiber cassumunar Roxb. ("Phlai" in Thai) has been used as an anti-asthmatic drug in Thai traditional medicine. However, the effect of this plant on mucin production has not been reported.

OBJECTIVE

The aim of the present study was to investigate whether crude ethanolic extracts of Zingiber cassumunar (CEZE) suppress phorbol12-myristate 13-acetate (PMA)-induced mucin production and gene expression in human airway epithelial cells and if so, to examine whether the suppression of mucin gene expression is mediated via the mitogen-activated protein kinase (MAPK) signal transduction pathways.

METHODS

Confluent NCI-H292 cells were pretreated with CEZE for 2 hours and then stimulated with 100 or 200 nmol/l PMA for 8 h. The levels of MUC2 and MUC5AC mRNA were determined by RT-PCR and real-time PCR. Levels of total mucin; MUC2 and MUC5AC inculture supernatants were measured using ELLA and ELISA assays, respectively. Extracellular signal-regulated kinase (ERK), JNK, p38 MAPK protein levels were analyzed by Western blotting.

RESULTS

CEZE (5-100?g/ml) significantly inhibited total mucin production, including MUC2 and MUC5AC mRNA and proteins induced by PMA in NCI-H292 cells. The extracts obviously inhibited the phosphorylation of ERK, but not JNK and p38 in PMA-stimulated NCI-H292 cells. Our results suggest that Z. cassumunar-mediated suppression of PMA-induced MUC2 and MUC5AC mRNA operates via ERK inhibition.

CONCLUSION

Z. cassumunar suppresses PMA-induced MUC2 and MUC5AC gene expression in human airway epithelial cells via inhibition of ERK MAPK-dependent pathway.

摘要

背景

黏蛋白的过度表达常常导致严重的气道病变。泰国姜黄(泰国语中的“Phlai”)的根茎在泰国传统医学中被用作抗哮喘药物。然而,这种植物对黏蛋白产生的影响尚未见报道。

目的

本研究旨在探究泰国姜黄乙醇粗提物(CEZE)是否能抑制佛波醇12-肉豆蔻酸酯13-乙酸酯(PMA)诱导的人气道上皮细胞中黏蛋白的产生及基因表达,若有此作用,则进一步研究黏蛋白基因表达的抑制是否通过丝裂原活化蛋白激酶(MAPK)信号转导途径介导。

方法

将汇合的NCI-H292细胞用CEZE预处理2小时,然后用100或200 nmol/l PMA刺激8小时。通过RT-PCR和实时PCR测定MUC2和MUC5AC mRNA的水平。分别使用ELLA和ELISA法测定培养上清液中总黏蛋白、MUC2和MUC5AC的水平。通过蛋白质印迹法分析细胞外信号调节激酶(ERK)、JNK、p38 MAPK蛋白水平。

结果

CEZE(5-100μg/ml)显著抑制NCI-H292细胞中由PMA诱导的总黏蛋白产生,包括MUC2和MUC5AC mRNA及蛋白。提取物明显抑制PMA刺激的NCI-H292细胞中ERK的磷酸化,但不影响JNK和p38的磷酸化。我们的结果表明,泰国姜黄介导的对PMA诱导的MUC2和MUC5AC mRNA的抑制作用是通过抑制ERK实现的。

结论

泰国姜黄通过抑制ERK MAPK依赖性途径抑制PMA诱导的人气道上皮细胞中MUC2和MUC5AC基因的表达。

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