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首页> 外文期刊>Life sciences >Nicotinic acetylcholine receptor subunits and receptor activity in the epithelial cell line HT29.
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Nicotinic acetylcholine receptor subunits and receptor activity in the epithelial cell line HT29.

机译：上皮细胞系HT29中的烟碱型乙酰胆碱受体亚基和受体活性。

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摘要

In the present study we have used RT-PCR to investigate nicotinic acetylcholine receptor (nAChR) subunit expression, and studied the effect of nicotine on TNFalpha-induced cytokine (IL-8) release in the epithelial cell line HT29. RNA was extracted using a commercial kit and amplified by RT-PCR. RT-PCR products were separated by electrophoresis and visualised using ethidium bromide. IL-8 release was measured by ELISA from cells activated for 6 h with TNFalpha (50 ng ml(-1)) in the absence and presence of nicotine (10(-11)-10(-6) M). HT29 cells contained mRNA for beta1, alpha4, alpha5, and alpha7 nAChR subunits. Activation of HT29 cells increased IL-8 release from undetectable amounts to 3.92 +/- 0.51 ng ml(-1) (n = 5). Nicotine significantly inhibited TNFalpha-induced IL-8 release in a concentration related manner with peak inhibition occurring at 10(-7) M (2.39 +/- 0.78 ng ml(-1), n = 5). Our data suggests that, while HT29 cells express mRNA for nAChR subunits, the only nAChR subunits that could form functional receptors and inhibit IL-8 release are alpha7.
机译：在本研究中，我们已使用RT-PCR研究烟碱型乙酰胆碱受体（nAChR）亚基表达，并研究了尼古丁对TNFα诱导的上皮细胞系HT29释放的细胞因子（IL-8）的影响。使用市售试剂盒提取RNA，并通过RT-PCR扩增。通过电泳分离RT-PCR产物，并使用溴化乙锭可视化。在不存在和存在尼古丁（10（-11）-10（-6）M）的情况下，通过ELISA从TNFalpha（50 ng ml（-1））活化6小时的细胞中测量IL-8的释放。 HT29细胞包含beta1，alpha4，alpha5和alpha7 nAChR亚基的mRNA。 HT29细胞的激活使IL-8释放量从不可检测的量增加到3.92 +/- 0.51 ng ml（-1）（n = 5）。尼古丁以浓度相关的方式显着抑制TNFalpha诱导的IL-8释放，在10（-7）M（2.39 +/- 0.78 ng ml（-1），n = 5）处出现峰抑制。我们的数据表明，虽然HT29细胞表达nAChR亚基的mRNA，但唯一可以形成功能受体并抑制IL-8释放的nAChR亚基是alpha7。

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