Protective Effect of Buyang Huanwu Decoction on Ang Ⅱ-induced Ferroptosis in H9c2 Cells Based on Nrf2/HO-1 Signaling Pathway
To investigate the regulatory effect of Buyang Huanwu Decoction on ferroptosis of H9c2 rat cardiomyocytes in the environment of angiotensin Ⅱ(Ang Ⅱ)which based on Nrf2/HO-l signaling pathway.Rat myocardial cells(H9c2)were cultured in vitro,and the normal group(ZC),model group(MX),low dosage group of Buyang Huanwu Decoction(BD),medium dosage group of Buyang Huanwu Decoction(BZ),high dosage group of Buyang Huanwu Decoction(BG)and positive control group(YX)were set up.All groups were treated with 1 μmol/L AngⅡ established a myocardial cell injury model except for the normal group,the intervention group of traditional Chinese medicine was respectively cultured in culture media containing 0.5,1 and 2 mg/mL Buyang Huanwu decoction,the YX was cultured with medium containing 10 μmol/L ferrostatin-1(Fer-1),and the cells were collected after 24 h.The activity of H9c2 cells in each group was assessed by CCK-8 assay.Cytometric iron content detection kit to determine the iron content of H9c2 cells.Immunofluorescence staining to detect the expression of Nrf2 protein in each group.The mRNA expression levels of nuclear transcription-related factor 2(Nrf2),hemored oxygenase-1(HO-1),and glutathione peroxidase 4(GPX4)were detected by q-PCR method in each group.ELISA kits was used to detect the protein expression levels of glutathione(GSH)and GPX4 in H9c2 cells.The Western blot method was used to detect the protein expression of Nrf2 and HO-1 in H9c2 cells.The results show that the high-dose groups of Buyang Huanwu Decoction significantly increases Ang Ⅱ-induced decrease in H9c2 cells viability(P＜0.01),and reduced iron content under Ang Ⅱ condition(P＜0.05).Compared with the model group,the BG significantly reversed the decline of gene and protein expression levels of GSH,GPX4,Nrf2 and HO-1 in H9c2 cells induced by AngⅡ,and increases the expression of Nrf2,which was inhibited under AngⅡ conditions,consistent with the positive control results(P＜0.01,P＜0.05).Buyang Huanwu Decoction may regulate H9c2 cells ferroptosis through Nrf2/HO-1 signaling pathway,thereby protecting myocardial injury secondary to hypertension.
万方数据
维普
