谷氨酰胺对细胞缺糖损伤的保护作用

Protective effect of glutamine on PC12 cells under glucose deprivation

目的观察谷氨酰胺(glutamine,Gln)对细胞缺糖(glucose desprivation,GD)损伤的保护作用,并探讨可能的作用机制。方法在以无糖培养大鼠肾上腺嗜铬细胞瘤(PC12)细胞建立细胞缺糖损伤模型的基础上,首先以噻唑蓝比色(MTT)法、流式细胞法、细胞线粒体跨膜电位检测等方法观察Gln对缺糖损伤细胞模型的作用;再以大鼠大脑中动脉线栓法建立动物脑缺血模型,观察Gln对动物缺血性损伤的保护作用;同时用免疫细胞化学法、Western blot印迹法检测细胞中应激基因葡萄糖调节蛋白75(grp75)的表达变化。结果Gln能使离体细胞在缺糖应激下存活率增加、凋亡率降低,线粒体跨膜电位稳定;动物实验显示Gln能减轻动物因缺血造成的脑损伤;免疫细胞化学法、Western blot印迹法检测结果表明Gln可以上调细胞中grp75的表达。结论Gln对细胞缺糖损伤和动物缺血性脑损伤具有一定的保护作用,这种保护作用可能与上调细胞中的应激基因grp75的表达有关。

Purpose To observe the protective effect of Glutamine （Gin） on Rat pheochromocytoma （PC12）cells inducible by glucose deprivation and on animal brain by ischemia, to explore its possible mechanism. Methods The in vitro and in vivo model were built to imitate human brain ischemia insuit. MTT and flow cytometry analysis were taken to detect the growth rate and apoptosis rate of PC12 cell. Neurological deficit score and infarcted volume were detected to estimate ischemic injuried brain of the animal model. The grp75 expression in PC12 cells with administration of Gln was analyzed by western blot and RT-PCR. Results When the cell was under glucose deprivation, treated with Gln, its survival rate increased and apoptosis rate decreased. Neurological deficit score and infarcted volume also documented that its protective effect of Gin on in vivo model was significant. Gin could upreglate the expression of grp75 in vitro model. Conclusions Gln could protect cells from the injury induced by glucose deprivation both in vitro and in vivo. The mechanism of these protective effect is related to the upregulation of expression of grp75.