Impact of atractylodin on ferroptosis in glioma cells by regulating the Nrf2/SLC7A11/GPX4 signaling pathway
Objective To investigate the impact of atractylodin on ferroptosis in glioma cells by regulating the nuclear factor E2-related factor 2(Nrf2)/solute carrier family 7a member 11(SLC7A11)/glutathione peroxidase 4(GPX4)signaling pathway and the mechanism.Methods The glioma cell line U251 was cultured in vitro and grouped into control group,low-dose atractylodin group(5mol/L),medium-dose atractylodin group(10mol/L),high-dose atractylodin group(20mol/L),and ML385 group(Nrf2 inhibitor 1mol/L).Cell proliferation was detected by MTT and EdU assay.Apoptotic rate was measured by flow cytometry.Relative levels of ferrous ions(Fe2+),lactate dehydrogenase(LDH),malondialdehyde(MDA),glutathione(GSH),and reactive oxygen species(ROS)were detected using commercial kits.Western blot was applied to detect the expression of Ki-67,cleared caspase3,Nrf2,SLC7A11,and GPX4 proteins.A nude mouse model of glioma was established,followed by the observation of tumor growth and expression levels of Ki-67,cleared caspase3,Nrf2,SLC7A11,and GPX4 in tumor tissues.Results In vitro results showed that,compared with those of the control group,U251 cells in the low,medium,and high-dose atractylodin groups presented significantly higher levels of Fe2+,MDA,LDH,ROS,apoptosis rate,and protein level of cleaved caspase3,but significantly lower level of GSH,absorbance at 490nm(OD490)(24h,48h),cell proliferation rate,and expression levels of Ki-67,Nrf2,SLC7A11,and GPX4(P<0.05).The above indexes were comparable between high-dose atractylodin group and ML385 group(P>0.05).In vivo results showed that compared with nude mice of the model group,those in the atractylodin group and ML385 group presented significantly lower tumor mass,tumor volume,and the expression levels of Ki-67,Nrf2,SLC7A11,and GPX4 proteins in the tumor tissue,but significantly higher expression level of cleaved caspase3(P<0.05).There were no significant differences in tumor mass,tumor volume,tumor inhibition rate,and protein expression levels between the atractylodin group and ML385 group(P>0.05).Conclusion Atractylodin may promote ferroptosis in glioma cells by inhibiting the Nrf2/SLC7A11/GPX4 signaling pathway.
atractylodinNrf2/SLC7A11/GPX4 signal pathwayglioma cellsferroptosis
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维普
