L-谷氨酰胺对PC12细胞中grp75的调控作用  

EFFECT OF GLUTAMINE ON THE EXPRESSION OF grp75 IN PC12 CELLS

作　　者：刘雯[1] 张艳[1] 吴青锋[1] 李锦燕[1] 左伋[1] 

机构地区：[1]复旦大学上海医学院细胞与遗传医学系,上海200032

出　　处：《实验生物学报》2005年第5期423-431,共9页Acta Biologiae Experimentalis Sinica

摘　　要：条件必需氨基酸谷胺酰胺可上调细胞中热激蛋白(hsp)的表达,为观察谷氨酰胺是否对hsp 家族成员grp75的表达具有调控作用,以PC12细胞为模型用免疫组化、蛋白质印迹法和RT-PCR 等方法检测谷胺酰胺对grp75基因的表达的影响;并以MTT法观察谷氨酰胺对PC12的细胞和grp75低表达的PC12细胞缺糖损伤的保护作用。结果表明谷氨酰胺可以上调grp75的表达,特别是对缺糖细胞的上调作用更显著;但这种上调作用与谷氨酰胺的作用浓度和作用时间并未显示出有明显的关系。MTT检测显示,谷氨酰胺使细胞在缺糖条件下的存活率明显上升;grp75低表达细胞与未转染的细胞相比这种保护效应明显降低。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 Gin, and glucose-free DMEM within Gin, 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 Gin can upregulated the expression of grp75 in PC 12 cell line, and the effect is more significantly in PC 12 cell which was glucose deprivation than the normal cell. To investigate the effect of Gin on PC12 cells under glucose deprivation, MTT method was used to monitor the cell viability after Gin 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 PC 12 cells is markedly decreased.

关 键 词：L-谷氨酰胺 葡萄糖调节蛋白 缺糖损伤 PC12细胞 GRP75 调控作用 条件必需氨基酸 蛋白质印迹法 保护作用 MTT法 

分 类 号：R363[医药卫生—病理学]