Mechanism of gypenosides on tau overexpression induced cell damage based on metabonomics
Objective To investigate the effect of gypenosides(GPs)on improving the abnormal cell metabolism induced by abnormal tau protein overexpression,and to explore the mechanism of GPs in the treatment of Alzheimer's disease.Methods Microtubule-associated protein tau(MAPT)overexpressing N2a cell were constructed by CRISPR/Cas9 technique.The effects of GPs on the survival ratio of N2a and MAPT cells were detected by Cell Counting Kit-8(CCK-8)to determine the safe dose range of GPs.N2a and MAPT cells were cultured in vitro,and different concentrations of GPs(50,100 μg·mL-1)were added to MAPT cells for intervention.After co-incubation for 24 h,the cells were collected and the total tau protein levels were detected by western blotting.Metabolomics technology was used to detect the effects of GPs on metabolites of MAPT cells and identify the different metabolites and pathways.Results GPs(50,100,200 μg·mL-1)had no significant effect on N2a cell viability.GPs(5,50,100,200 μg·mL-1)had no significant effect on MAPT cell viability.Compared with N2a control group,the tau protein level in MAPT cells was significantly increased(P＜0.001),and the total tau protein content in MAPT cells was significantly decreased by 50 μg·mL-1 GPs.The metabolomics results showed that the different metabolites enriched in lipid and lipid molecules,organic acids,nucleosides,nucleotides and analogues,etc.KEGG pathway analysis demonstrated differential metabolites were mainly concentrated in nucleotide metabolism,amino acid synthesis,glycerol phospholipid metabolism and other pathways.After GPs treatment,the level of CDP-Choline and phosphocholine(PC)were increased(P＜0.001),while the contents of glycerophosphocholine(GPC)and glycerin-3-phosphoethanolamine were significantly decreased(P＜0.001).Conclusion GPs can attenuate the abnormal overexpression of tau protein,and regulate the metabolism pathway of glycerophospholipid by increasing CDP-Choline and PC,reducing GPC and sn-Glycerol-3-phosphoethanolamine.
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