Journal: Cell Death & Disease

Article Title: SIRT1 deficiency promotes age-related heart failure through enhancing ferroptosis via GATA4-HADHA-GPX4 axis

doi: 10.1038/s41419-026-08634-z

Figure Lengend Snippet: A Neonatal rat cardiomyocytes were transfected with negative control siRNA (si-NC) or siRNA targeting SIRT1 (si-SIRT1). Representative immunoblots and quantitative analysis of SIRT1, HADHA, transferrin (TF), ferritin heavy chain 1 (FTH1), and glutathione peroxidase 4 (GPX4) protein levels are shown ( n = 5 per group). B In silico molecular docking analysis illustrating the predicted interaction between SIRT1 and HADHA. C Relative mRNA expression of Hadha in cardiomyocytes following SIRT1 silencing, as determined by quantitative real-time PCR ( n = 6 per group). D Bioinformatic enrichment analysis identifying GATA4 as a putative transcription factor associated with HADHA regulation, based on the ChEA Transcription Factor Targets and Binding Site Profiles datasets. E Co-immunoprecipitation analysis demonstrating the interaction between SIRT1 and GATA4 in cardiomyocytes. Cell lysates were immunoprecipitated with anti-SIRT1 antibody, followed by immunoblotting with antibodies against SIRT1 and GATA4. Representative immunoblots are shown. F Schematic illustration of the HADHA promoter region showing the predicted GATA4-binding motif (−327 to −337 bp) and the serial 5′-truncated promoter constructs generated for luciferase reporter assays. ( G ) Dual-luciferase reporter assays assessing the transcriptional activity of individual serially truncated HADHA promoter constructs following GATA4 overexpression or control treatment ( n = 6 per group). H Experimental design for resveratrol intervention in aging rats. Eighteen-month-old rats were randomly assigned to receive vehicle (Aging Control) or resveratrol (Aging RES) by daily gavage for 4 months and were analyzed at 22 months of age. I Representative immunoblots of SIRT1, HADHA, and GATA4 protein expression in cardiac tissues from Aging Control and Aging RES rats. J Representative immunoblots of TF, FTH1, and GPX4 protein expression in cardiac tissues from Aging Control and Aging RES rats. K Quantitative analysis of SIRT1, HADHA, GATA4, TF, FTH1, and GPX4 protein levels in cardiac tissues from Aging Control and Aging RES rats ( n = 6 per group). L Representative M-mode echocardiographic images from Aging Control and Aging RES rats. M Left ventricular ejection fraction (EF) in Aging Control and Aging RES rats ( n = 6 per group). N Left ventricular fractional shortening (FS) in Aging Control and Aging RES rats ( n = 6 per group). O Serum NT-proBNP levels in Aging Control and Aging RES rats ( n = 6 per group). P Representative hematoxylin and eosin (H&E)–stained sections of left ventricular tissue. Scale bars, 50 μm. Q Representative Masson trichrome–stained sections of left ventricular tissue. Scale bars, 50 μm. R Quantification of left ventricular collagen volume fraction ( n = 6 per group). The data are given as mean ± SEM and compared by Student’s t test.

Article Snippet: The membranes were then incubated with primary antibodies against TF (1:500, Proteintech, cat#17435-1-AP, RRID: AB_2035023), FTH1 (1:500, Bioss, cat#bs-5907R, RRID: AB_11050926), GPX4 (1:500, Abcam, cat#ab125066, RRID: AB_10973901), HADHA (1:2000, Proteintech, cat#10758-1-AP, RRID: AB_2115593), and SIRT1 (IP, 1:500, Proteintech, cat#60303-1-Ig, RRID: AB_2881417; IB, 1:500, Abcam, cat#ab189494, RRID: AB_2864311), GATA4 (1:500, Proteintech, cat#19530-1-AP, RRID: AB_10642003), and GAPDH (1:10000, Abcam, cat#ab128915, RRID: AB_2747414) at 4°C overnight.

Techniques: Transfection, Negative Control, Western Blot, In Silico, Expressing, Real-time Polymerase Chain Reaction, Binding Assay, Immunoprecipitation, Construct, Generated, Luciferase, Activity Assay, Over Expression, Control, Staining

Journal: Cell Death & Disease

Article Title: SIRT1 deficiency promotes age-related heart failure through enhancing ferroptosis via GATA4-HADHA-GPX4 axis

doi: 10.1038/s41419-026-08634-z

Figure Lengend Snippet: Aging is associated with reduced cardiac SIRT1 expression. SIRT1 deficiency attenuates GATA4 binding and transcriptional activation of the HADHA promoter, resulting in reduced HADHA expression. Downregulation of HADHA impairs mitochondrial function and increases reactive oxygen species (ROS) accumulation. Excessive ROS results in glutathione depletion and suppression of GPX4, thereby promoting lipid peroxidation and ferroptotic cell death in cardiomyocytes. Ferroptosis-driven myocardial injury ultimately contributes to the development of age-related heart failure.

Article Snippet: The membranes were then incubated with primary antibodies against TF (1:500, Proteintech, cat#17435-1-AP, RRID: AB_2035023), FTH1 (1:500, Bioss, cat#bs-5907R, RRID: AB_11050926), GPX4 (1:500, Abcam, cat#ab125066, RRID: AB_10973901), HADHA (1:2000, Proteintech, cat#10758-1-AP, RRID: AB_2115593), and SIRT1 (IP, 1:500, Proteintech, cat#60303-1-Ig, RRID: AB_2881417; IB, 1:500, Abcam, cat#ab189494, RRID: AB_2864311), GATA4 (1:500, Proteintech, cat#19530-1-AP, RRID: AB_10642003), and GAPDH (1:10000, Abcam, cat#ab128915, RRID: AB_2747414) at 4°C overnight.

Techniques: Expressing, Binding Assay, Activation Assay