GLT-1基因敲低抑制头孢曲松介导的对APP/PS1阿尔茨海默病小鼠认知缺陷、GLT-1和xCT表达及活性的改善作用。
GLT-1 Knockdown Inhibits Ceftriaxone-Mediated Improvements on Cognitive Deficits, and GLT-1 and xCT Expression and Activity in APP/PS1 AD Mice.
作者信息
Gao JunXia, Liu LiZhe, Liu Chao, Fan ShuJuan, Liu LiRong, Liu ShuFeng, Xian Xiao-Hui, Li Wen-Bin
机构信息
Department of Pathophysiology, Neuroscience Research Center, Hebei Medical University, Shijiazhuang, China.
Hebei Key Lab of Laboratory Animal Science, Laboratory Animal Center, Hebei Medical University, Shijiazhuang, China.
出版信息
Front Aging Neurosci. 2020 Oct 6;12:580772. doi: 10.3389/fnagi.2020.580772. eCollection 2020.
OBJECTIVE
Glutamate transporter-1 (GLT-1) and system x mediate glutamate uptake and release, respectively. Ceftriaxone has been reported to upregulate GLT-1 expression and improve cognitive decline in APP/PS1 mice. The aim of the present study was to elucidate the role of GLT-1 in ceftriaxone-mediated improvement on cognitive deficits and associated changes in xCT (catalytic subunit of system x ) expression and activity using GLT-1 knockdown APP/PS1 mice.
METHODS
GLT-1 knockdown (GLT-1) mice were generated in C57BL/6J mice using the CRISPR/Cas9 technique and crossed to APP/PS1 mice to generate GLT-1APP/PS1 mice. The cognition was evaluated by novel object recognition and Morris water maze tests. GLT-1 and xCT expression, GLT-1 uptake for glutamate, and glutathione levels of hippocampus were assayed using Western blot and immunohistochemistry, H-glutamate, and glutathione assay kit, respectively.
RESULTS
In comparison with wild-type mice, APP/PS1 mice exhibited significant cognitive deficits, represented with poor performance in novel object recognition and Morris water maze tests, downregulated GLT-1 expression and glutamate uptake. Ceftriaxone treatment significantly improved the above impairments in APP/PS1 mice, but had negligible impact in GLT-1APP/PS1 mice. The xCT expression increased in APP/PS1 and GLT-1APP/PS1 mice. This upregulation might be a compensatory change against the accumulated glutamate resulting from GLT-1 impairment. Ceftriaxone treatment restored xCT expression in APP/PS1 mice, but not in GLT-1APP/PS1 mice. Glutathione levels decreased in APP/PS1 mice in comparison to the wild-type group. After ceftriaxone administration, the decline in glutathione level was restored in APP/PS1 mice, but not in GLT-1APP/PS1 mice.
CONCLUSION
Ceftriaxone improves cognitive impairment of APP/PS1 mice by upregulating GLT-1-mediated uptake of glutamate and co-regulation of GLT-1 and xCT in APP/PS1 mice.
目的
谷氨酸转运体1（GLT-1）和x系统分别介导谷氨酸的摄取和释放。据报道，头孢曲松可上调GLT-1表达并改善APP/PS1小鼠的认知衰退。本研究的目的是利用GLT-1基因敲低的APP/PS1小鼠，阐明GLT-1在头孢曲松介导的认知缺陷改善中的作用以及xCT（x系统的催化亚基）表达和活性的相关变化。
方法
使用CRISPR/Cas9技术在C57BL/6J小鼠中构建GLT-1基因敲低（GLT-1）小鼠，并与APP/PS1小鼠杂交，以生成GLT-1APP/PS1小鼠。通过新物体识别和莫里斯水迷宫试验评估认知功能。分别使用蛋白质免疫印迹法、免疫组织化学法、H-谷氨酸和谷胱甘肽检测试剂盒检测海马体中GLT-1和xCT的表达、GLT-1对谷氨酸的摄取以及谷胱甘肽水平。
结果
与野生型小鼠相比，APP/PS1小鼠表现出明显的认知缺陷，在新物体识别和莫里斯水迷宫试验中表现不佳，GLT-1表达下调，谷氨酸摄取减少。头孢曲松治疗显著改善了APP/PS1小鼠的上述损伤，但对GLT-1APP/PS1小鼠的影响可忽略不计。APP/PS1和GLT-1APP/PS1小鼠的xCT表达增加。这种上调可能是对GLT-1损伤导致的谷氨酸积累的一种代偿性变化。头孢曲松治疗可恢复APP/PS1小鼠的xCT表达，但对GLT-1APP/PS1小鼠无效。与野生型组相比，APP/PS1小鼠的谷胱甘肽水平降低。头孢曲松给药后，APP/PS1小鼠谷胱甘肽水平的下降得到恢复，但GLT-1APP/PS1小鼠未恢复。
结论
头孢曲松通过上调GLT-1介导的谷氨酸摄取以及共同调节APP/PS1小鼠中的GLT-1和xCT来改善APP/PS1小鼠的认知障碍。
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