氯胺酮对星形胶质细胞葡萄糖吸收的作用及其机制  

Effect of ketamine on glucose uptake in astrocytes and its machanism

作　　者：陈嘉鑫 冯建国 贾静 刘行 王晓斌 CHEN Jiaxin;FENG Jianguo;JIA Jing;LIU Xing;WANG Xiaobin(Department of Anesthesiology,Affiliated Hospital of Southwest Medical University,Luzhou 646000,China)

机构地区：[1]西南医科大学附属第一医院麻醉科,泸州646000

出　　处：《山西医科大学学报》2020年第3期247-252,共6页Journal of Shanxi Medical University

基　　金：国家自然科学基金资助项目(81271478)。

摘　　要：目的探讨氯胺酮对人星形胶质细胞葡萄糖吸收、葡萄糖转运蛋白(glucose transporters,GLUTs)及相关信号通路的影响。方法以培养的正常人星形胶质细胞株(HA1800)为研究对象,将细胞分为50μmol/L氯胺酮处理0(对照组),0.5,3,6,12,24 h,作用结束后加入荧光标记2-脱氧葡萄糖(2-NBDG)检测HA1800细胞的葡萄糖吸收情况。将细胞分别用0(对照组),10,25,50,100,200μmol/L的氯胺酮处理6 h,作用结束后加入2-NBDG检测细胞葡萄糖吸收情况。将细胞分别用0(对照组),10,25,50,100μmol/L的氯胺酮处理6 h,处理结束后用Western blot检测GLUTs,同时检测ERK1/2、AKT、AMPK信号通路的表达。将细胞分别用0(对照组)和50μmol/L氯胺酮作用6 h后免疫荧光观察p-ERK1/2细胞内分布。结果与0 h比较,50μmol/L氯胺酮作用6 h时增加HA1800细胞的葡萄糖吸收(P=0.018)。与0μmol/L比较,25,50,100μmol/L氯胺酮处理6 h后,HA1800细胞的葡萄糖吸收显著增加(P 25μmol/L=0.0033,P 50μmol/L=0.0001,P 100μmol/L=0.0074)。与0μmol/L比较,50μmol/L氯胺酮作用6 h后GLUT3(P=0.014)以及p-ERK1/2(P=0.0069)蛋白表达明显增加。与0μmol/L比较,免疫荧光检测显示50μmol/L氯胺酮组p-ERK1/2从细胞质向细胞核聚集明显。结论氯胺酮可能通过ERK1/2信号通路促进人正常星形胶质细胞葡萄糖吸收和GLUT3表达增加。Objective To examine the effects of ketamine on glucose uptake,glucose transporters(GLUTs)and related signaling pathways in astrocytes.Methods The normal human astrocyte cell line HA1800 cells were cultured with 50μmol/L ketamine for 0,0.5,3,6,12,24 h,respectively,and then the glucose uptake was detected by 2-[N-(7-Nitrobenz-2-oxa-1,3-diazol-4-yl)Amino]-2-Deoxyglucose(2-NBDG)uptake assay.The HA1800 cells were treated with different concentrations of ketamine(0,10,25,50,100,200μmol/L),and then the glucose uptake was detected by 2-NBDG uptake assay.The HA1800 cells were treated with different concentrations of ketamine(0,10,25,50,100,200μmol/L)for 6 h,and then the expression levels of GLUTs and ERK1/2,AKT,AMPK signaling pathways were measured by Western blot analysis.The HA1800 cells were treated with different concentrations of ketamine(0,50μmol/L)for 6 h,and then the location of p-ERK1/2 in the astrocytes was detected by immunofluorescence technique.Results Compared with 0 h,50μmol/L ketamine for 6 h increased the glucose uptake(P=0.018).Compared with 0μmol/L,25,50,100μmol/L ketamine significantly increased the glucose uptake(P 25μmol/L=0.0033,P 50μmol/L=0.0001,P 100μmol/L=0.0074).Compared with 0μmol/L,50μmol/L ketamine for 6 h significantly increased the expression levels of GLUT3(P=0.014)and p-ERK1/2 in HA1800(P=0.0069).Compared with 0μmol/L,50μmol/L ketamine for 6 h induced translocation of p-ERK1/2 into nucleus from cytoplasm.Conclusion Ketamine may promote the glucose uptake and the expression level of GLUT3 through ERK1/2 signaling pathway.

关 键 词：氯胺酮 葡萄糖吸收 星形胶质细胞 ERK1/2信号通路 

分 类 号：R965[医药卫生—药理学]