作　　者： (郑俊超）; (畅洪昇）; (马素亚）; (程丹）; (刘永刚）; (刘婧玮）; (鲁艺）;

机构地区： 北京中医药大学中医学院

出　　处： 《中华中医药杂志》 2017年第8期3717-3720,共4页

摘　　要： 目的:基于ERK1/2通路探讨巴戟天多糖提取物(MHP)、菟丝子多糖提取物(CCP)对H_2O_2损伤SHSY-5Y细胞的保护作用。方法:采用SH-SY-5Y细胞为神经细胞体外研究模型,CCK8法检测不同浓度(50、100、200、400、800、1 600mg/L)MHP和CCP对细胞活力的影响和对H_2O_2损伤SH-SY-5Y细胞活力的影响,分为对照组、H_2O_2组、MHP+H_2O_2组、CCP+H_2O_2组、H_2O_2+MHP+CCP(1∶1)组。运用Western blot方法检测PSD-95、p-ERK和ERK等神经系统相关蛋白表达情况,分为:对照组、H_2O_2组、MHP+H_2O_2组、CCP+H_2O_2组、H_2O_2+MHP+CCP(1∶1)组和对照组、PD98059组、PD98059+MHP+CCP组、MHP+CCP组。结果:CCK8检测结果显示,与对照组比较MHP、CCP 1 600mg/L可以显著提高SH-SY-5Y细胞活力,对H_2O_2损伤SH-SY-5Y细胞有明显保护作用,其中MHP+CCP+H_2O_2(1∶1)组抗氧化损伤能力最强。Western blot结果显示,与对照组比较,H_2O_2可显著抑制PSD-95表达(P<0.05)。与模型组比较,MHP和CCP可显著促进PSD-95表达(P<0.05),其中MHP+CCP(1∶1)+H_2O_2组PSD-95表达量最高(P<0.05)。MHP+CCP(1∶1)组可显著促进p-ERK表达(P<0.05)。与MPH+CCP组比较,ERK1/2阻断剂PD98059可显著抑制MHP+CCP(1∶1)组的作用(P<0.05)。结论:MHP+CCP(1∶1)应用对H_2O_2损伤SH-SY-5Y细胞PSD的保护作用最好,可能与促进PSD-95蛋白和p-ERK1/2通路蛋白表达有关。 Objective: To study the protective effect of Chinses herb morida pfficinalis how polysaccharide(MHP) and cuscuta chinensis polysaccharide(CCP) on H_2O_2-induced injury on SY-SY-5Y cells basic of ERK1/2 pathway. Methods: Take SH-SY-5Y cells as neuron model in vitro, SH-SY-5Y cells were treated with different concentrations of MHP and CCP(50, 100, 200, 400, 800, 1 600mg/L) and divided the cells into control group, H_2O_2 group, MHP+H_2O_2 group, CCP+H_2O_2 group, H_2O_2+MHP+CCP(1:1) group, using CCK8 methods to detect the cell viability. Using Western blot method to detect the expression of nervous system related protein PSD-95, p-ERK and ERK. Divided the cells into control group, H_2O_2 group, MHP+ H_2O_2 group, CCP+H_2O_2 group, H_2O_2+MHP+ CCP(1:1) group and control group, PD98059 group, PD98059+MHP+CCP group, MHP+CCP group. Results: The CCK8 results has shown that MHP and CCP can significantly improve cell viability at a concentration of 1 600mg/L, and protect SH-SY-5Y cells from H_2O_2 injury, compared with control group(P<0.05). Among them, the MHP+CCP(1:1) group appears the most antioxidant effect. The Western blot results has shown that H_2O_2 decreased the expression of PSD-95, compared with control group(P<0.05). MHP, CCP and MHP+CCP(1:1) group increased the expression of PSD-95, among them, the expression of PSD-95 of MHP+CCP(1:1) group is the highest, compared with model group(P<0.05). The antagonist of ERK1/2 PD98059 block the expression of p-ERK of MHP+CCP(1:1) group, compared with MHP+CCP group(P<0.05). Conclusion: Both MHP and CCP can exert protective effect on H_2O_2 injured SH-SY-5Y cells, but MHP +CCP(1:1) group works best. And this antioxidant effect could be related to increasing expression of PSD-95 and p-ERK.

关 键 词： 巴戟天多糖 菟丝子多糖 抗氧化 突触后致密蛋白 细胞外信号调节激酶