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首页> 外文期刊>Biomedical Research >Effect of five taste ligands on the release of CCK from an enteroendocrine cell line, STC-1
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Effect of five taste ligands on the release of CCK from an enteroendocrine cell line, STC-1

机译:五个味觉配体对肠内分泌细胞STC-1释放CCK的影响

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摘要

Here, we investigated which taste ligand induces the CCK (cholecystokinin) release from intestinal STC-1 cells. We first developed a new assay to measure the release of CCK. The expression vector for CCK type A receptor (CCKAR) was permanently introduced into HEK293T cells and a cell line was established (CCKAR/HEK). Then, STC-1 cells were treated with taste ligands and the incubated buffer of STC-1 cells containing released CCK was applied to CCKAR/HEK cells. Since CCKAR couples to Gq-signaling, the CCK-induced receptor activation can be monitored by the method of Ca~(2+)-imaging. Therefore, when CCK is released from STC-1 cells to culture medium with taste stimulation, Ca~(2+) activation of CCKAR/HEK should be observed. Among five different taste ligands (saccharin, Na-glutamate, NaCl, denatonium benzoate, HCl), only denatonium benzoate and HCl induced the release of CCK in STC-1 cells. Thus, we found that only specific taste ligands induce the CCK release, and that other three taste ligands cannot induce the release of CCK despite of their ability to elevate the intracellular Ca~(2+) level in STC-1 cells.
机译:在这里,我们研究了哪种味觉配体诱导了肠STC-1细胞释放的CCK(胆囊收缩素)。我们首先开发了一种新的方法来测量CCK的释放。将用于CCK A型受体的表达载体(CCKAR)永久导入HEK293T细胞,并建立细胞系(CCKAR / HEK)。然后,用味觉配体处理STC-1细胞,并将含有释放的CCK的STC-1细胞的孵育缓冲液应用于CCKAR / HEK细胞。由于CCKAR耦合到Gq信号,可以通过Ca〜(2 +)-成像方法监测CCK诱导的受体激活。因此,当通过刺激味觉将CCK从STC-1细胞释放到培养基中时,应观察到CCKAR / HEK的Ca〜(2+)活化。在五个不同的味觉配体(糖精,谷氨酸钠,NaCl,苯甲酸地那铵,HCl)中,只有苯甲酸地那铵和HCl诱导了STC-1细胞中CCK的释放。因此,我们发现只有特定的味觉配体诱导了CCK的释放,而其他三种味觉配体尽管能够提高STC-1细胞中细胞内Ca〜(2+)的水平,却无法诱导CCK的释放。

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