谷氨酸刺激引起反应性星形胶质细胞分泌CSPGs增加的实验研究  

Experimental study of glutamic acid induced CSPGs secretion increase in A1 astrocytes

作　　者：陈虹瑛 马瑗锾 曾湘[2] 曾园山[2] 兰月 CHEN Hong-ying;MA Yuan-huan;ZENG Xiang;ZENG Yuan-shan;LAN Yue(Department of Rehabilitation Medicine,The Second Affiliated Hospital,South China University of Technology,Guangzhou,Guangdong Province 510180,China;Department of Histology and Embryology,Zhongshan School of Medicine,Sun Yat-sen University,Guangzhou,Guangdong Province 510080,China;Guangzhou Institute of Clinical Medicine,South China University of Technology,Guangzhou,Guangdong Province 510180,China;Guangzhou Key Laboratory of Aging Frailty and Neurorehabilitation,Guangzhou,Guangdong Province 510180,China)

机构地区：[1]华南理工大学附属第二医院,康复医学科,广东广州510180 [2]中山大学中山医学院组织学与胚胎学教研室,广东广州510080 [3]华南理工大学附属第二医院,广州临床医学研究所,广东广州510180 [4]广州市老年虚弱与神经康复重点实验室,广东广州510180

出　　处：《解剖学研究》2023年第2期97-102,共6页Anatomy Research

基　　金：国家自然科学基金重大项目(81891003);国家自然科学基金项目(81974357);广州市科技计划项目(202206010197);广州市第一人民医院科研启动基金(KYQD20210017)。

摘　　要：目的 探讨谷氨酸对A1型星形胶质细胞分泌硫酸软骨素蛋白多糖(CSPGs)的影响。方法提取小鼠原代星形胶质细胞培养并将其诱导为A1型星形胶质细胞。用浓度梯度谷氨酸作用于A1型星形胶质细胞72 h后,采用MTT法检测谷氨酸随浓度增加对A1型星形胶质细胞活力的影响、ELISA检测不同浓度谷氨酸及谷氨酸拮抗剂作用于A1型星形胶质细胞72 h后细胞上清中CSPGs的含量。结果 (1)原代培养的星形胶质细胞GFAP阳性率达98%以上;(2)经诱导后原代培养的星形胶质细胞转变为C3阳性的A1型星形胶质细胞;(3)谷氨酸对于A1型星形胶质细胞的活力抑制作用随谷氨酸浓度的升高而增大;(4)经不同浓度谷氨酸作用72 h后,A1型星形胶质细胞上清中CSPGs含量增加,且呈现剂量依赖性;谷氨酸对A1型星形胶质细胞的这种抑制作用可以被谷氨酸受体拮抗剂所抵消。结论 谷氨酸浓度增加会降低A1型星形胶质细胞活力并刺激其分泌CSPGs;通过谷氨酸受体阻断剂可以拮抗谷氨酸对A1型星形胶质细胞的影响。Objective To explore the effect of glutamate on the secretion of CSPG by Al reactive astrocytes and whether glutamate stimulate the secretion of CSPG by reactive astrocytes.Methods Primary astrocytes were cultured and induced into Al reactive astrocytes.The concentration gradient glutamate was used to treat Al reactive astrocytes for 72 h,and the toxic effect of glutamate on the reactive astrocytes with increasing concentration was detected by MTT assay(represented by proliferation inhibition rate).The content of CSPGs in the supernatant of A1 reactive astrocytes was determined by ELISA assay after dfferent concentrations of glutamate and glutamate antagonists were treated for 72 h.Results ①The positive rate of GFAP in primary cultured astrocytes was more than 98%.②Primary cultured astrocytes could be induced to be C3 positive Al reactive astrocytes after induction.③The cytotoxicity of glutamate to Al reactive astrocytes increased with the increase of glutamate concentration;④The content of CSPGs in the supernatant of A1-reactive astrocytes increased in a dose-dependent manner after 72 h treatment with different concentrations of glutamate,and this effect could be offset by glutamate receptor antagonists.Conclusion Excessive glutamate on Al reactive astrocytes could stimulate the secretion of CSPGs,and glutamate receptor block-ers can antagonize the stimulation of glutamate on A1 reactive astrocytes.

关 键 词：谷氨酸 A1型星形胶质细胞 硫酸软骨素蛋白多糖 细胞培养 

分 类 号：R338.1[医药卫生—人体生理学]