Conggan Zhixin(从肝治心)Decoction Regulates Ferroptosis in H9c2 Cardiomyocytes Exposed to Hypoxia/Reoxygenation Injury via Nrf2/HO-1 Pathway
Objective:To investigate the effects of Conggan Zhixin(从肝治心)Decoction on ferroptosis of H9c2 cardiomyocytes via the nuclear factor E2-related factor 2(Nrf2)/heme oxygenase-1(HO-1)signaling pathway.Methods:H9c2 cardiomyocytes were exposed to hypoxia/reoxygenation(H/R)to establish a cell model of myocardial ischemia-reperfusion injury(MIRI).Cells were classified into normal control,model control,15％blank serum control group,26.6 g/kg Conggan Zhixin Decoction-containing serum(15％),NK252(20 μmol/L),ML385(5 μmol/L),and 26.6 g/kg Conggan Zhixin Decoction-containing serum(15％)+ML385(5 μmol/L)groups.The Cell Counting Kit-8(CCK-8)was used to examine cell viability.DCFH-DA,TMRE,and FerroOrange probes were used to measure the levels of reactive oxygen species(ROS),mitochondrial membrane potential,and intracellular Fe2+.western blotting and Real-time reverse transcription polymerase chain reaction(RT-qPCR)were employed to determine the protein levels and mRNA of Nrf2,HO-1,hephaestin(Heph),divalent metal transporter 1(DMT1),and acyl-CoA synthetase long-chain family member 4(ACSL4).Results:Compared with the normal control group,the model control group showed decreased mitochondrial membrane potential,elevated ROS and Fe2+levels,down-regulated expression of Nrf2,HO-1,and Heph,and up-regulated expression of DMT1 and ACSL4(P＜0.05,P＜0.01).Compared with the model control group,26.6 g/kg Conggan Zhixin Decoction-containing serum(15％)and NK252(20 μmol/L)groups showed increased mitochondrial membrane potential,lowered ROS and Fe2+levels,up-regulated expression of Nrf2,HO-1,and Heph,and down-regulated expression of DMT 1 and ACSL4(P＜0.05,P＜0.01).Compared with the ML385(5 μmol/L)group,the 26.6 g/kg Conggan Zhixin Decoction-containing serum(15％)+ML385(5 μmol/L)group showed increased mitochondrial membrane potential,lowered ROS and Fe2+levels,up-regulated expression of Nrf2,HO-1,and Heph,and down-regulated expression of DMT1 and ACSL4(P＜0.05,P＜0.01).Conclusion:Conggan Zhixin Decoction may attenuate ferroptosis in H9c2 cardiomyocytes exposed to H/R injury by modulating the Nrf2/HO-1 signaling pathway.
NETL
NSTL
万方数据
