谷氨酰胺对PC12细胞grp75表达的影响

Shi yan sheng wu xue bao Pub Date : 2005-10-01
Wen Liu, Yan Zhang, Qing Feng Wu, Jin Yan Li, Ji Zuo
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引用次数: 0

摘要

谷氨酰胺(Gln)是一种条件必需氨基酸,可能是热应激反应的潜在增强剂。葡萄糖调节蛋白75(grp75)是hsp家族的一员。为研究谷氨酰胺对grp75表达的影响,采用DMEM、无葡萄糖DMEM、Gln内DMEM和Gln内无葡萄糖DMEM培养PC12细胞,采用免疫细胞化学和western blot检测grp75表达,逆转录-聚合酶链反应(RT-PCR)检测细胞内grp75 mRNA水平。数据表明,Gln可上调PC12细胞株中grp75的表达,且在葡萄糖剥夺的PC12细胞中的作用比正常细胞更为显著。为了研究Gln对葡萄糖剥夺PC12细胞的影响,采用MTT法监测Gln处理后葡萄糖剥夺细胞的细胞活力。实验结果表明,0.2 ~ 40 mmol/L浓度范围内的谷氨酰胺显著提高了无糖DMEM细胞的活力。grp75低表达对PC12细胞的保护作用明显降低。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
[Effect of glutamine on the expression of grp75 in PC12 cells].

Glutamine (Gln), a conditionally essential amino acid, can be a potential enhancer of the heat stress response. And glucose-regulated protein 75(grp75) is a member of the hsp family. To evaluate the effect of glutamine on the expression of grp75, PC12 cell was cultured with DMEM, glucose-free DMEM, DMEM within Gln, and glucose-free DMEM within Gln, and the expression of grp75 was detected by immunocytochemistry and western blot, the mRNA level of grp75 in the cells was detected by reverse transcriptase-polymerase chain reaction (RT-PCR). Data indicated that Gln can upregulated the expression of grp75 in PC12 cell line, and the effect is more significantly in PC12 cell which was glucose deprivation than the normal cell. To investigate the effect of Gln on PC12 cells under glucose deprivation, MTT method was used to monitor the cell viability after Gln treatment for the cell under glucose depriving. The experiments results showed that glutamine at a concentration range of 0.2-40 mmol/L significantly enhanced the cell viability in glucose-free DMEM. And the protective effect to grp75 low-expresssion on PC12 cells is markedly decreased.

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