Effects of Panaxadiol on Tau Protein Phosphorylation and Fyn/GluN 2B Signaling Pathway in APP-SH-SY 5Y Cells
- VernacularTitle:人参二醇对APP-SH-SY5Y细胞中Tau蛋白磷酸化及Fyn/GluN2B信号通路的影响
- Author:
Xicai LIANG
1
;
Ying LIN
2
;
Honghe XIAO
2
;
Liang KONG
2
;
Jingxian YANG
2
Author Information
1. Laboratory Animal Center,Liaoning University of TCM,Shenyang 110847,China
2. College of Pharmacy,Liaoning University of TCM,Liaoning Dalian 116600
- Publication Type:Journal Article
- Keywords:
Panaxadiol;
APP-SH-SY5Y cells;
Tau protein;
Phosphorylation;
Fyn/GluN2B signaling pathway
- From:
China Pharmacy
2021;32(12):1485-1491
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVE:To study the effects and mechanism of panaxadiol (PD) on Tau protein phosphorylation in the SH-SY5Y cells transfected with APP gene(APP-SH-SY5Y). METHODS :The target of PD and non-receptor tyrosine kinases Fyn was verified by molecular docking. SH-SY 5Y cells were cultured in vitro ,and the APP-SH-SY 5Y cell models and green fluorescent (GFP)-SH-SY5Y cell model (control cell )was constructed. The expression of Aβ1-42 was detected so as to verify the success of APP-SH-SY5Y cell model. Taking GFP-SH-SY 5Y cells as control ,the effects of 5,10,20,30,40 μmol/L PD and 125,250, 500,1 000,2 000 nmol/L PP 2(Fyn inhibitor ,positive control )on the survival rate of APP-SH-SY 5Y cells were detected by CCK-8 assay after treated for 24 h,so as to confirm the optimal concentration. The concentration of Ca 2 + ,the ratio fophosphorylated Tau protein (p-Tau)/Tau,phosphorylatedn Src(p-Src)/Fyn and phosphorylated glutamate receptor 2B(p-GluN2B)/ GluN2B were detected in APP-SH-SY 5Y cells after trated with the optimal concentration of PD and PP 2 for 24 h. RESULTS :The results of molecular simulation docking showed that PD could target Fyn protein. Compared with GFP-SH-SY 5Y cells ,the protein expression of Aβ1-42 in APP-SH-SY 5Y cell were increased significantly (P<0.01). The optimal concentration of PD and PP 2 were 20 μmol/L and 500 nmol/L. The 20 μmol/L PD and 500 nmol/L PP 2 could increase the survival rate of the cells and reduced the concentration of Ca 2+,the ratio of p-Tau/Tau ,p-Src/Fyn,and p-GluN 2B/GluN2B. CONCLUSIONS:PD can reduce the the phosphorylation of Tau protein through inhibiting Fyn/GluN 2B signaling pathway.