Department of Physiology, School of Medicine, National Yang-Ming University, Taipei, Republic of China.
J Physiol Pharmacol. 2013 Oct;64(5):535-43.
The purpose of the present study was to explore the effect of arecoline on phytohemagglutinin (PHA)-stimulated interleukin-2 (IL-2) secretion, the expression of alpha7-nicotinic acetylcholine receptors (α7-nAChRs), prostaglandin E2(PGE2) protein, and IL-2 mRNA in human lymphocyte cells (Jurkat cell line). The IL-2 and PGE2 were determined by enzyme-linked immunosorbent assay (ELISA). The expressions of phosphorylated extracellular signal-regulated kinase (ERK) and α7-nAChRs were determined by Western blotting. The level of IL-2 mRNA was determined by reverse-transcriptase polymerase chain reaction (RT-PCR). Arecoline, in a dose-dependent manner, significantly decreased IL-2 and PGE2 secretion by Jurkat cells incubated with 0 or 5 μg/ml 5 μg/ml PHA. PGE2 also significantly inhibited IL-2 secretion by Jurkat cells in a dose-dependent manner. In addition, reduced expression of PHA-induced ERK phosphorylation was observed in Jurkat cells treated with arecoline. PHA-enhanced IL-2 mRNA expression was also inhibited by arecoline. These results imply that arecoline inhibits the release of PGE2 and PHA-induced IL-2 secretion by Jurkat cells and that these effects seem to occur, at least in part, either through the attenuation of ERK in conjunction with a decrease of PHA-induced IL-2 mRNA expression. These results imply that arecoline inhibits the protein expression of α7-nAChRs , the release of PGE2 and PHA-induced IL-2 secretion by Jurkat cells.
本研究旨在探讨槟榔碱对植物血球凝集素(PHA)刺激白细胞介素-2(IL-2)分泌、α7-烟碱型乙酰胆碱受体(α7-nAChRs)、前列腺素 E2(PGE2)蛋白和 IL-2 mRNA 在人淋巴细胞(Jurkat 细胞系)中表达的影响。通过酶联免疫吸附测定(ELISA)测定 IL-2 和 PGE2 的表达。通过 Western blot 测定磷酸化细胞外信号调节激酶(ERK)和α7-nAChRs 的表达。通过逆转录聚合酶链反应(RT-PCR)测定 IL-2 mRNA 的水平。槟榔碱呈剂量依赖性显著降低了 Jurkat 细胞与 0 或 5 μg/ml PHA 孵育时的 IL-2 和 PGE2 分泌。PGE2 也呈剂量依赖性显著抑制 Jurkat 细胞中 IL-2 的分泌。此外,在槟榔碱处理的 Jurkat 细胞中观察到 PHA 诱导的 ERK 磷酸化表达减少。槟榔碱还抑制 PHA 增强的 IL-2 mRNA 表达。这些结果表明,槟榔碱抑制 PGE2 的释放和 PHA 诱导的 Jurkat 细胞中 IL-2 的分泌,这些作用似乎至少部分通过与 PHA 诱导的 IL-2 mRNA 表达减少相关的 ERK 衰减来发生。这些结果表明,槟榔碱抑制 α7-nAChRs 的蛋白表达、Jurkat 细胞中 PGE2 的释放和 PHA 诱导的 IL-2 分泌。