Effects of huwentoxin-I on catecholamines in cultured PC12 cells
Effects of huwentoxin-I on catecholamines in cultured PC12 cells
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DOI：
10.3109/15376516.2015.1008287
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发表时间：
2015-01-01
影响因子：
3.2
通讯作者：
Wang, Xianchun
中科院分区：
文献类型：
--
作者：
Chen, Jia;Dai, Zhipan;Wang, Xianchun
生物样品中的儿茶酚胺的测量仍然是当前神经递质释放研究中的分析挑战，尽管已经致力于相关方法学的研究。本文首先建立了一种基于荧光的儿茶酚胺定量测定方法，然后将其应用于研究肽类神经毒素虎纹伊蚊毒素-I（HWTX-I）对培养的PC 12细胞中儿茶酚胺的影响。结果显示，不同浓度（10、50、100和1000 nM）的HWTX-1处理20 min显著增加培养基中多巴胺（DA）含量（p < 0.01），在最高测试浓度下与对照相比增加了约1.6倍。在20 min内，培养基中DA含量随处理时间的延长而增加。与此相反，HWTX-1减少去甲肾上腺素（NE）的含量在培养基中。而HWTX-Ⅰ处理PC 12细胞后，细胞内DA和NE含量保持在相对恒定的水平。这些数据表明，HWTX-Ⅰ对神经内分泌细胞的DA和NE有不同的影响，增强DA的作用，减弱NE的作用。
The measurement of catecholamines in biological samples remains a current analytical challenge in the study on neurotransmitter release, although much effort has been devoted toward the relevant methodology. Here we first established a fluoresence-based method for the quantitative determination of catecholamines and then applied it to the investigation of effects of huwentoxin-I (HWTX-I), a peptide neurotoxin, on catecholamines in cultured PC12 cells. The results showed that treatment with HWTX-I at different concentrations (10, 50, 100 and 1000 nM) for 20 min significantly increased dopamine (DA) content in the culture medium (p < 0.01), with similar to 1.6-fold increase compared with that of the control at the highest concentration tested. Within 20 min, DA content in the medium increased with increasing the treatment time. In contrast, HWTX-I decreased norepinephrine (NE) content in the medium. However, after the PC12 cells were treated with HWTX-I, the intracellular contents of both DA and NE were kept at a relatively constant level. These data suggest that HWTX-I differentially affects the DA and NE of the neuroendocrine cells, with enhancing the actions of DA and weakening those of NE.