三七总皂苷对氧糖剥夺损伤的小鼠星形胶质细胞谷氨酸摄取的影响  

Effect of Panax Notoginseng Saponins on Glutamate Uptake in Astrocytes of Mice with Oxygen Glucose Deprivation Injury

作　　者：乔园 穆庆丽 吴启航 杨洁[1] QIAO Yuan;MU Qingli;WU Qihang;YANG Jie(Institute of Chinese Materia Medica,Shaanxi Provincial Academy of Traditional Chinese Medicine,Xi’an 710002,China)

机构地区：[1]陕西省中医药研究院中药研究所,陕西西安710002

出　　处：《中医药学报》2023年第8期37-41,共5页Acta Chinese Medicine and Pharmacology

基　　金：陕西省自然科学基金项目(2019JQ-995);陕西省中医医院基金项目(2020-01)。

摘　　要：目的:探讨三七总皂苷对氧糖剥夺/复氧(OGD/R)诱导的星形胶质细胞谷氨酸摄取能力的影响。方法:体外培养ICR小鼠大脑皮层星形胶质细胞,分为正常组、模型组(OGD 6 h/R 24 h)和三七总皂苷组(OGD 6 h/R 24 h+20μg·mL^(-1))。免疫细胞荧光染色鉴定GFAP标记星形胶质细胞;采用CCK-8法检测星形胶质细胞活力,同时检测细胞外液LDH的释放;采用生化法检测细胞Na^(+)-K^(+)-ATPase活性、细胞谷氨酸摄取率;Western Blot法和RT-PCR法检测细胞GLT-1、GS mRNA表达及蛋白水平变化。结果:经特异性蛋白GFAP鉴定,细胞呈阳性表达。OGD/R损伤后,星形胶质细胞活力降低,LDH释放增多(P<0.01)。细胞对谷氨酸的摄取率明显降低(P<0.01),Na^(+)-K^(+)-ATPase活性降低,同时GLT-1和GS mRNA表达和蛋白表达水平显著下降(P<0.05)。三七总皂苷干预后,细胞活力明显增强(P<0.05),LDH释放明显减少(P<0.01)。星形胶质细胞对谷氨酸的摄取能力增强(P<0.01),Na^(+)-K^(+)-ATPase活性升高(P<0.01),GLT-1和GS蛋白表达和mRNA表达水平升高(P<0.01)。结论:三七总皂苷抑制OGD/R引起谷氨酸浓度升高,可能是通过提高Na^(+)-K^(+)-ATPase活性,上调GLT-1和GS的表达,从而增强星形胶质细胞对谷氨酸的清除能力。Objective:To investigate the effects of Panax Notoginseng saponins(PNS)on glutamate uptake in oxygen glucose deprivation/reoxygention(OGD/R)induced astrocytes.Methods:Primary ICR mice cerebral cortex astrocytes were cultured in vitro and divided into the normal group,the model group(OGD 6 h/R 24 h)and the PNS group(20μg·mL^(-1)).GFAP-labeled astrocytes were identified by immunocytocfluorescence staining;CCK-8 asssy was used to detect the cell viability,and the release of extracellular LDH was measured.The astrocytes Na^(+)-K^(+)-ATPase activity and the glutamate uptake rate were detected by biochemical method;Western Blot and RT-PCR were used to detect the protein and mRNA expressions of GLT-1 and GS.Results:The cultured astrocytes were identified by GFAP immunofluorescence staining,the cells were positive fluorescence staining.After OGD/R injury,cell viability was significantly decreased,LDH release was obviously increased(P<0.01).The uptake rate of glutamate was decreased significantly(P<0.01),Na^(+)-K^(+)-ATPase activity was decreased,and the protein and mRNA expressions of GLT-1 and GS were obviously down-regulated(P<0.05).PNS remarkably increased cell viability(P<0.05),and significantly reduced LDH release(P<0.01).Glutamate uptake was significantly enhanced(P<0.01),Na^(+)-K^(+)-ATPase activity was obviously increased(P<0.01),and the protein and mRNA expressions of GLT-1 and GS were significantly up-regulated(P<0.01).Conclusion:PNS inhibits OGD/R-induced increase of glutamate via increasing activity of Na^(+)-K^(+)-ATPase and up-regulating expressions of GLT-1 and GS,thus to strengthen the scavenging activity of astrocyte to glutamate.

关 键 词：三七总皂苷 星形胶质细胞 氧糖剥夺 谷氨酸摄取 

分 类 号：R285.5[医药卫生—中药学]