This study was designed to investigate the effect of eriodictyol on glioma cell autophagy and its role in glioma autophagy by regulating the Akt/mTOR pathway.In this essay,CCK-8 assay and colony formation were conducted to evaluate the effects of eriodictyol on proliferation.The glioma cell lines with stable GFP-mRFP-LC3B expression were constructed to observe the autophagic flux.After treatment of eriodictyol (100 μmol/L) for 48 h,autophagosomes were observed by fluorescent microscopy.Then,respectively treated with eriodictyol (100 μmol/L),rapamycin (200 nmol/L),or eriodictyol (100 μmol/L) + rapamycin (200 nmol/L) for 48 h,glioma cell lines were observed using fluorescent microscopy to ensure autophagic flux.The autophagy and AKT/mTOR pathway-related proteins were detected by immunoblotting.Moreover,A model of glioma subcutaneously transplant in nude mice was constructed to study the effects of eriodictyol on the autophagy of glioma in vivo.The results showed that the proliferation of glioma cells was significantly inhibited by eriodictyol in a dose and time-dependent manner (P < 0.05).The formation of autophagosome and the expressions of LC3Ⅱ,Beclin 1,ATG5 proteins were enhanced by eriodictyol.After treatment with eriodictyol,the glioma cells expressing GFP-mRFP-LC3B appeared bright yellow.Eriodictyol weakened the effect of rapamycin on autophagic flux and increased P62 expression in glioma cells.Furthermore,the inhibition of p-Akt and p-mTOR proteins by eriodictyol in glioma cells was rescued by Akt agonist 740 Y-P.In vivo,similar results were obtained as in vitro. In summary,the proliferation and autophagic flux in glioma cells was inhibited by eriodictyol.The mechanism of eriodictyol affecting autophagy might be through the downregulation of Akt/mTOR pathway.