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.%Aim To investigate the protective effects of supercritical CO2 fluid extract(SFE)of Notoginseng a-gainst glutamate-induced PC12 cells damage and the underlying mechanism. Methods PC12 cells were dealt with glutamate to establish cell models. MTT as-say,LDH method,Hoschst 33342 staining,Fluo-3 /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 re-ceptors 2 (GluR2),respectively. Results Glutamate was cytotoxic to PC12 cells with an inhibitory concen-tration 50(IC 50 )of 25 mmol·L - 1 . Pretreatment with SFE(25,50,100 mg·L-1)and FSC231(100 μmol ·L-1 )and SFE(100 mg·L-1 )+FSC231(100μmol ·L-1 )remarkablely improved cell viability,reduced LDH leakage,decreased apoptosis rate,debased intra-cellular calcium concentration,decreased the expres-sion of PICK1 ,and increased the expression of GluR2 . Conclusions SFE of Notoginseng shows protective effects against glutamate-induced PC12 cell damage, and its mechanism may be related to the inhibition of PICK1 and the increase of GluR2 protein expression.
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