Protective effect of dihydromyricetin against hippocampal neuronal injury caused by iron overload and its mechanism
Objective To investigate the protective effect of dihydromyricetin(DMY)against injury of primary hip-pocampal neurons caused by ferric ammonium citrate(FAC)and its mechanism.Methods Primary hippocampal neurons were collected from 24-hour neonatal Sprague-Dawley rats for in vitro culture,and CCK-8 assay was used to measure the viability of pri-mary hippocampal neurons treated with different concentrations of DMY and determine the administration concentration of DMY.Primary hippocampal neurons were divided into control group(H group),100 mmol/L DMY treatment group(I group),250 mmol/L FAC treatment group(J group),and 100 mmol/L DMY+250 mmol/L FAC treatment group(K group).Flow cy-tometry was used to measure the content of reactive oxygen species(ROS)in each group;colorimetry was used to measure the content of malondialdehyde(MDA)in each group;Western blot was used to measure the protein expression levels of nuclear factor erythroid 2-related factor 2(Nrf2),heme oxygenase-1(HO-1),and glutathione peroxidase 4(GPX4)in each group.Results CCK-8 assay showed the highest viability of primary hippocampal neurons at the concentration of 100 mmol/L for DMY(F=9.95,P＜0.05),and therefore,this concentration was used for subsequent experiments.Compared with the H group,the J group had significant increases in the content of ROS and MDA(F=176.81,5 523.35,P＜0.05),and compared with the J group,the K group had significant reductions in the content of ROS and MDA(F=18.21,412.96,P＜0.05).Compared with the H group,the J group had significant reductions in the relative protein expression levels of Nrf2,HO-1,and GPX4(F=27.35-81.32,P＜0.05),and compared with the J group,the K group had significant increases in the relative protein expression levels of Nrf2,HO-1,and GPX4(F=8.74-21.46,P＜0.05).Conclusion DMY can alleviate oxidative stress damage in primary hippocampal neurons due to iron overload,possibly by activating the Nrf2-HO-1/GPX 4 pathway in primary hippocampal neurons.
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