Study on some proteins controling cell death in oxidative stress-induced neuronal cell death.

我们已经报道谷氨酸通过抑制胱氨酸摄取发挥细胞毒性作用，导致细胞谷胱甘肽（GSH）水平显著降低，使细胞暴露于氧化应激导致细胞死亡。这种细胞死亡似乎是伴随1-2Mbp巨大DNA断裂导致核小体间DNA断裂的细胞凋亡。我们通过对C6大鼠胶质瘤细胞提取物的细胞内肽进行二维凝胶电泳（2D-DIGE）检测细胞死亡调节蛋白或因子，扩展了最近对细胞内蛋白的研究。我们鉴定了一种蛋白激酶C-δ是参与谷氨酸诱导的细胞死亡的蛋白激酶之一，此外，在谷氨酸处理的细胞中，用LC-MS/MS鉴定了从10种蛋白中提取的3种肽。在谷氨酸处理的胶质瘤细胞中，78-kDa葡萄糖调节蛋白（GRP78）、膜联蛋白- a5和半乳糖凝集素-1等三种肽增加。

We have reported that glutamate exerts cytotoxic action through inhibition of cystine uptake, which leads to a marked decrease in cellular glutathione (GSH) levels exposing the cells to oxidative stresses leading to cell death. This cell death seemed to be apoptosis accompanying with 1-2Mbp giant DNA fragmentation led to internucleosomal DNA fragmentation. We expand on recent studies of intracellular proteins by examining the cell death regulating proteins or factors by using two-dimensional differential in-gel electrophoresis (2D-DIGE) in the intracellular peptides from cell extracts of C6 rat glioma cells. We identified a protein kinase C-δas one of protein kinases involved in the glutamate-induced cell death, and furthermore, identified 3 peptides derived from 10 proteins with LC-MS/MS in theglutamate-treated cells. Three peptides such as 78-kDa glucose-regulated protein (GRP78), annexin -A5and galectin-1 were increased in glutamate -treated glioma cells.