论文部分内容阅读
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 investigate the protective effect of Morus alba polysaccharide extract (MHP) and cotyledon polysaccharide extract (CCP) on H 2 O 2 -deficient SHSY-5Y cells based on ERK1 / 2 pathway. Methods: SH-SY-5Y cells were used as nerve cell model in vitro. CCK8 assay was used to detect the effects of MHP and CCP at different concentrations (50,100,200,400,800,1 600mg / L) on cell viability and H 2 O 2 injury SH SY-5Y cells were divided into control group, H 2 O 2 group, MHP + H 2 O 2 group, CCP + H 2 O 2 group and H 2 O 2 + MHP + CCP group. Western blot was used to detect the expression of PSD-95, p-ERK, ERK and other nervous system related proteins in the control group, H 2 O 2 group, MHP + H 2 O 2 group, CCP + H 2 O 2 group, H 2 O 2 + MHP + CCP ) Group and control group, PD98059 group, PD98059 + MHP + CCP group and MHP + CCP group. Results: The results of CCK8 assay showed that compared with the control group, MHP and CCP1 600 mg / L could significantly increase the viability of SH-SY-5Y cells and protect the SH-SY-5Y cells from H 2 O 2 -induced injury. MHP + CCP + H 2 O 2 1: 1) group, the strongest antioxidant capacity. Western blot results showed that, compared with the control group, H 2 O 2 significantly inhibited the expression of PSD-95 (P <0.05). Compared with the model group, MHP and CCP significantly promoted the expression of PSD-95 (P <0.05), and the highest expression of PSD-95 in MHP + CCP (1: 1) + H 2 O 2 group was the highest (P <0.05). MHP + CCP (1: 1) group can significantly promote the expression of p-ERK (P <0.05). Compared with MPH + CCP group, ERK1 / 2 blocker PD98059 significantly inhibited the effect of MHP + CCP (1: 1) group (P <0.05). CONCLUSION: The protective effect of MHP + CCP (1: 1) on PSD of SH-SY-5Y cells is best, which may be related to the promotion of PSD-95 protein and p-ERK1 / 2 pathway protein expression.