Protective effect of taxifolin on H2O2-induced H9C2 cell pyroptosis / 中南大学学报(医学版)

ABSTRACT

Objective:To explore the effect of taxifolin on H2O2-induced pyroptosis in H9C2 cells and the possible mechanisms.Methods:The H9C2 cells was divided into 3 groupsa control group,a hydrogen peroxide (H2O2) group and a taxifolin group.The morphology of H9C2 cells was observed by inverted phase contrast microscope.The mitochondrial membrane potential was measured by JC-1 staining and flow cytometry.The alteration of the level of reactive oxygen species (ROS) was detected by specific mitochondrial probe.The protein levels of cysteinyl aspartate specific proteinase-1 (caspase-1) was determined by Western blot.The mRNA levels of interleukin-18 (IL-18),interleukin-1a (IL-1a),interleukin-1b (IL-1b),absent in melanoma 2 (AIM2),apoptosis-associated apeck-like protein (ASC),nucleotide-binding oligomerization domain-like receptor protein 3 (NLRP3)and nucleotide-binding oligomerization domain-like receptor family caspase recruitment domaincontaining protein 4 (NLRC4) were determined by reverse transcription-polymerase chain reaction (RT-PCR).Results:Compared with the control group,the morphology of H9C2 cells obviously changed in the H2O2-treated group,which was guadually improved in the presence of taxifolin.Compared with the control group,the mitochondrial membrane potential was markedly decreased in the H2O2-treated cells,accompanied by the increase of ROS (both P＜0.05).Compared with the H2O2 group,the mitochondrial membrane potential changes in the taxifolin group was increased while the ROS was decreased,with significant difference (both P＜0.05).Compared with the control group,the protein level of caspase-1 and the mRNA levels of IL-18,IL-1a,IL-1b,AIM2,ASC,NLRP3 and NLRC4 in the H2O2-treated group were significantly increased (all P＜0.05),which were attenuated in the presence of taxifolin (all P＜0.05).Conclusion:Taxifolin can protect H9C2 cells from oxidative injury,and it is able to suppress the H2O2-induced H9C2 cell pyroptosis through inhibition of AIM2,NLRP3 and NLRC4 in flammasome.