三七超临界CO_2萃取物对谷氨酸损伤PC12细胞的保护作用  

Protective effects of SFE-CO_2 of Notoginseng against glutamate-induced apoptosis in PC12 cells

作　　者：段贤春 周安[1] 彭代银 鲍金云 夏伦祝 DUAN Xian chun;ZHOU An;PENG Dai yin;BAO Jin yun;XIA Lun zhu(School of Pharmacy, Anhui University of Chinese Medicine the First Affiliated Hospital of Anhui University of Chinese Medicine, Hefei230038,China;Dept of Pharmacy the First Affiliated Hospital of Anhui University of Chinese Medicine, Hefei230038,China)

机构地区：[1]安徽中医药大学药学院,安徽合肥230038 [2]安徽中医药大学第一附属医院药学部,安徽合肥230038

出　　处：《中国药理学通报》2017年第12期1685-1690,共6页Chinese Pharmacological Bulletin

基　　金：安徽省自然科学基金资助项目(No 1508085QH191);安徽高校省级自然科学研究重点项目(No KJ2013A169);安徽省省级中医发展专项经费项目(No 2014ZYZJ01;2014ZYZJ02);安徽中医药大学校级探索性科研项目(No2016ts038)

摘　　要：目的研究三七超临界CO_2萃取物(SFE)对谷氨酸损伤PC12细胞的保护作用,并探讨其可能作用机制。方法以谷氨酸损伤PC12细胞为模型,采用MTT法检测细胞存活率,LDH法检测乳酸脱氢酶的漏出率,Hoechst 33342染色法检测细胞凋亡,Fluo-3/AM荧光染色法检测细胞内钙离子浓度,Western blot法检测PICK1、GluR2蛋白的表达。结果谷氨酸对PC12细胞具有兴奋性毒性作用,其半数抑制浓度(IC50)为25 mmol·L^(-1)。不同浓度三七SFE(25、50、100 mg·L^(-1))、PICK1抑制剂FSC231(100μmol·L^(-1))、三七SFE(100 mg·L^(-1))+FSC231(100μmol·L^(-1))预处理可明显提高细胞存活率,减少LDH的释放,降低PC12细胞的凋亡率,减少钙离子内流,降低PICK1蛋白表达,增加GluR2蛋白表达。结论三七超临界CO_2萃取物对谷氨酸损伤后的PC12细胞具有保护作用,机制可能与抑制PICK1、增加GluR2蛋白表达有关。AimTo investigate the protective effects of supercritical CO2fluid extract(SFE)of Notoginseng against glutamate induced PC12cells damage and the underlying mechanism.MethodsPC12cells were dealt with glutamate to establish cell models.MTT assay,LDH method,Hoschst33342staining,Fluo3/AM fluorescence staining and Western blot were used to observe the changes of cell viability,intracellular Ca2+concentration and the expression of protein that interacted with C kinase l(PICK1)and glutamate receptors2(GluR2),respectively.ResultsGlutamate was cytotoxic to PC12cells with an inhibitory concentration50(IC50)of25mmol·L-1.Pretreatment with SFE(25,50,100mg·L-1)and FSC231(100μmol·L-1)and SFE(100mg·L-1)+FSC231(100μmol·L-1)remarkablely improved cell viability,reduced LDH leakage,decreased apoptosis rate,debased intracellular calcium concentration,decreased the expression of PICK1,and increased the expression of GluR2.ConclusionsSFE of Notoginseng shows protective effects against glutamate induced PC12cell damage,and its mechanism may be related to the inhibition of PICK1and the increase of GluR2protein expression.

关 键 词：三七超临界CO2萃取物 谷氨酸 PC12细胞 钙离子 PICK1 GLUR2 

分 类 号：R284.1[医药卫生—中药学] R329.24[医药卫生—中医学]