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通过佛波酯诱导 Jurkat 细胞中组氨酸脱羧酶表达增强组胺产生。

Enhanced histamine production through the induction of histidine decarboxylase expression by phorbol ester in Jurkat cells.

机构信息

Graduate School of Life and Environmental Sciences, Faculty of Life and Environmental Sciences, Life Science Center of Tsukuba Advanced Research Alliance, University of Tsukuba, Tsukuba, Ibaraki 305-8577, Japan.

出版信息

Mol Med Rep. 2012 Nov;6(5):944-8. doi: 10.3892/mmr.2012.1049. Epub 2012 Aug 27.

Abstract

Histamine (HA), a mediator of inflammation, type I allergic responses and neurotransmission, is synthesized from L-histidine, the reaction of which is catalyzed by histidine decarboxylase (HDC). HDC has been reported to be induced by various stimuli, not only in mast cells and basophils, but also in T lymphocytes and macrophages. Although its mRNA has been shown to be increased in Jurkat cells when treated with phorbol 12-myristate 13-acetate (TPA), little is known concerning the induced production of HA by HDC. The present study quantified the trace amounts of intracellular HA using ultra-high liquid chromatography in combination with the 6-aminoquinoline carbamate-derivatization technique. To test whether the cellular level of HA is elevated by the induction of HDC in Jurkat cells treated with TPA, the peak corresponding to authentic HA in the cell lysate was fractioned and its molecular weight determined by matrix-assisted laser desorption/ionization quadrupole ion trap time-of-flight mass spectrometry. The results of this study show that the HA level is increased by the induction of HDC expression by TPA in Jurkat cells. Therefore, this method is useful in elucidating the physiological significance of HA production.

摘要

组胺 (HA) 是一种炎症介质、I 型过敏反应和神经递质,由 L-组氨酸合成,其反应由组氨酸脱羧酶 (HDC) 催化。已经报道 HDC 可被各种刺激诱导,不仅在肥大细胞和嗜碱性粒细胞中,而且在 T 淋巴细胞和巨噬细胞中也是如此。尽管已经表明 Jurkat 细胞在用佛波醇 12-肉豆蔻酸 13-乙酸酯 (TPA) 处理时其 mRNA 增加,但对于 HDC 诱导的 HA 产生知之甚少。本研究使用超高效液相色谱法结合 6-氨基喹啉氨基甲酸酯衍生化技术定量检测细胞内 HA 的痕量。为了测试 TPA 处理的 Jurkat 细胞中 HDC 诱导是否会升高细胞内 HA 的水平,将细胞裂解物中与真实 HA 对应的峰分离,并通过基质辅助激光解吸/电离四极杆离子阱飞行时间质谱法确定其分子量。本研究结果表明,TPA 诱导 HDC 表达可增加 Jurkat 细胞中 HA 的水平。因此,该方法可用于阐明 HA 产生的生理意义。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/4617/3493046/4ddbf42c0828/MMR-06-05-0944-g00.jpg

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