TRPC1 protects human SH-SY5Y cells against salsolinol-induced cytotoxicity by inhibiting apoptosis
TRPC1 protects human SH-SY5Y cells against salsolinol-induced cytotoxicity by inhibiting apoptosis
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DOI：
10.1016/j.brainres.2006.04.104
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发表时间：
2006-07-12
期刊：
影响因子：
2.9
通讯作者：
Singh, Brij B.
中科院分区：
文献类型：
--
作者：
Bollimuntha, Sunitha;Ebadi, Manuchair;Singh, Brij B.
猪毛菜醇是一种内源性神经毒素，可能参与帕金森病的发病机制。在这项研究中，我们试图确定是否salsolinol诱导的细胞毒性SH-SY 5 Y人神经母细胞瘤细胞，克隆细胞系表达多巴胺能活性，可以防止过表达的钙离子通道，瞬时受体电位（TRPC 1）蛋白。SH-SYSY细胞暴露于500 μ M salsolinol 12小时导致毒胡萝卜素或卡巴胆碱介导的Ca 2+内流显着减少。与这些结果一致，用猪毛菜醇处理的SH-SY 5 Y细胞显示TRPC 1蛋白水平降低约60%。共聚焦显微镜还显示，用猪毛菜醇处理的SH-SYSY细胞的TRPC 1蛋白质的质膜染色显著减少。有趣的是，TRPC 1的过表达增加了TRPC 1蛋白水平，也保护了SH-SY 5 Y神经母细胞瘤。通过3，[4，5-二甲基噻唑-2-基]-2，5-二苯基溴化四氮唑（MTT）测定法测定细胞对salsolinol介导的细胞毒性的抵抗。TRPC 1的保护作用被TRPC 1阻断剂镧或2APB阻断。通过毒胡萝卜素或卡巴胆碱激活TRPC 1蛋白进一步保护SH-SY 5 Y细胞免受猪毛菜醇处理。用凋亡标记物（YO-PRO-1）染色SH-SY 5 Y细胞显示TRPC 1蛋白保护免于凋亡。此外，TRPC 1过表达还抑制细胞色素c的释放，并降低细胞凋亡所需的BAX蛋白水平。总之，这些发现表明，在细胞表面TRPC 1蛋白表达的减少，响应于salsolinol可能是一个促成因素的多巴胺能神经元的细胞毒性。此外，TRPC 1的过表达可以抑制凋亡复合物，从而增加帕金森病神经细胞的存活率。(c)2006 Elsevier B. V.保留所有权利。
Salsolinol, an endogenous neurotoxin, may be involved in the pathogenesis of Parkinson's disease. In this study, we sought to determine whether salsolinol-induced cytotoxicity in SH-SY5Y human neuroblastoma cells, a cloned cell line which expresses dopaminergic activity, could be prevented by overexpressing a Ca2+ channel, transient receptor potential (TRPC1) protein. Exposure of SH-SYSY cells to 500 mu M salsolinol for 12 h resulted in a significant decrease in thapsigargin or carbachol-mediated Ca2+ influx. Consistent with these results, SH-SY5Y cells treated with salsolinol showed approximately 60% reduction in TRPC1 protein levels. Confocal microscopy also showed that SH-SYSY cells treated with salsolinol had a significant decrease in the plasma membrane staining of the TRPC1 protein. Interestingly, overexpression of TRPC1 increases TRPC1 protein levels and also protected SH-SY5Y neuroblastoma. cells against salsolinol-mediated cytotoxicity as determined by 3, [4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide (MTT) assay. The protective effect of TRPC1 was blocked by the addition of TRPC1 blockers lanthanum, or 2APB. Activation of TRPC1 protein by either thapsigargin or carbachol further protected SH-SY5Y cells from salsolinol treatments. Staining of SH-SY5Y cells with an apoptotic marker (YO-PRO-1) showed that TRPC1 protein protects against apoptosis. Furthermore, TRPC1 overexpression also inhibited cytochrome c release and decreased BAX protein levels required for apoptosis. Taken together, these findings suggest that the reduction in cell surface TRPC1 protein expression in response to salsolinol may be a contributory factor in cellular toxicity of the dopaminergic neurons. Furthermore, overexpression of TRPC1 could inhibit apoptotic complex thereby increasing neuronal cell survivability in Parkinson's disease. (c) 2006 Elsevier B.V. All rights reserved.