瘦素对PC12细胞中钠钙交换体的影响。
Effects of Leptin on Na+/Ca2+ Exchanger in PC12 Cells.
作者信息
Zhang Zijuan, Sun Shuguang, Du Caixia, Li Wei, Zhang Juan, Zhu Yanqin, Liu Peilin, Xing Ying
机构信息
Experimental teaching center, Basic Medical College of Henan University of Chinese Medicine, Zhengzhou, China.
出版信息
Cell Physiol Biochem. 2016;40(6):1529-1537. doi: 10.1159/000453203. Epub 2016 Dec 21.
BACKGROUND/AIMS: Alzheimer's disease (AD) is known to be related to alterations in neuronal intracellular calcium activity ([Ca2+]i). The present study revealed the distinct role of leptin in Na+/Ca2+-exchanger activity.
METHODS
[Ca2+]i was determined utilizing Fura-2 fluorescence. The activity of NCX was measured by removal of extracellular Na+ in the presence of external Ca2+. Na+/Ca2+-exchanger activity was further quantified from whole cell currents following removal of extracellular Na+. Na+/Ca2+-exchanger isoform NCX1 transcript levels and protein abundance were quantified by RT-PCR and Western blotting, respectively.
RESULTS
Exposure of PC12 cells to 30 µM amyloid (Aβ42) increased [Ca2+]i, an effect significantly blunted by 6 hours incubation with leptin before Aβ42 treatment. Moreover, leptin treatment significantly increased Na+/Ca2+-exchanger mediated Ca+ transport and current, NCX1 transcript level as well as NCX1 membrane protein abundance.
CONCLUSION
We show that leptin blunts Aβ42-evoked [Ca2+]i increase by increasing expression and activity of Na+/Ca2+-exchanger NCX1.
背景/目的：已知阿尔茨海默病（AD）与神经元细胞内钙活性（[Ca2+]i）的改变有关。本研究揭示了瘦素在钠/钙交换体活性中的独特作用。
方法
利用Fura-2荧光测定[Ca2+]i。在细胞外存在Ca2+的情况下，通过去除细胞外Na+来测量钠钙交换体（NCX）的活性。在去除细胞外Na+后，根据全细胞电流进一步量化钠/钙交换体活性。分别通过逆转录聚合酶链反应（RT-PCR）和蛋白质免疫印迹法（Western blotting）对钠/钙交换体亚型NCX1的转录水平和蛋白丰度进行量化。
结果
将PC12细胞暴露于30μM淀粉样蛋白（Aβ42）会增加[Ca2+]i，在Aβ42处理前用瘦素孵育6小时可显著减弱这种效应。此外，瘦素处理显著增加了钠/钙交换体介导的Ca+转运和电流、NCX1转录水平以及NCX1膜蛋白丰度。
结论
我们发现，瘦素通过增加钠/钙交换体NCX1的表达和活性，减弱了Aβ42诱发的[Ca2+]i升高。
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