Journal: Antioxidants

Article Title: Limitations of Ferroptosis Inhibitors on the Doxorubicin-Induced Cardiotoxicity

doi: 10.3390/antiox15010027

Figure Lengend Snippet: Ferroptosis inhibitors fail to rescue cardiac dysfunction. ( A ) Brief schedule of in vivo experiment. C57BL/6J mice were treated with ferrostatin-1 (Fer-1) or liproxstatin-1 (Lip-1) 24 h before doxorubicin injection and were dosed every 24 h. A single dose of doxorubicin was administered intraperitoneally once on day 1. Experimental groups: Vehicle, DOX (20 mg/kg, i.p.), DOX + Fer-1 (2 mg/kg, i.p.), and DOX + Lip-1 (10 mg/kg, i.p.) ( B ) The survival graph of mice from different groups was shown by Kaplan–Meier survival analysis. ( C ) Three days after intraperitoneal injection of doxorubicin, the mouse cardiac function was analyzed with echocardiography. The echocardiography data showed decreased fractional shortening and ejection fraction in mice treated with doxorubicin and Fer-1 or Lip-1. Survival rate, n = 5 per group; ECG, n = 5 per group. ( D ) Bar graph representation of the heart weight to body weight ratio of mice. ( E ) 3,3’-Diaminobenzidine immunohistochemical staining for 4-hydroxynonenal (4-HNE) in the mouse heart tissue; scale bars, 200 µm (left panel). Quantification of 4-HNE-specific 3,3’-Diaminobenzidine staining (right panel). ( F ) Bar graph representation of the N-terminal pro B-type natriuretic peptide (NT-proBNP) of mice. ( G ) Masson’s trichrome and Sirius red staining of fibrosis in the mouse heart (left panel) and quantification (right panel). ( H ) Western blot of ferroptosis cell death markers in mouse heart tissues treated with doxorubicin and Fer-1 or Lip-1. heart weight to body weight, n = 3 per group; 4-HNE, n = 3 per group; NT-proBNP, n = 7 per group; Masson’s trichrome and Sirius red, n = 3 per group; Western blots, n = 3 per group. Data are presented as mean ± S.D., where n represents the number of individual mice per group. Statistical significance was determined by one-way ANOVA followed by Sidak’s multiple comparison test for panels ( C , F ), and the Kruskal–Wallis test was used for panels ( D , E , G ), ns, no significance, * p < 0.05, ** p < 0.01, *** p < 0.005, **** p < 0.001.

Article Snippet: ECG recordings were conducted for 22 days, with each mouse housed individually in a cage equipped with a receiver (RPC-1, Data Sciences International).

Techniques: In Vivo, Injection, Immunohistochemical staining, Staining, Western Blot, Comparison

Journal: Antioxidants

Article Title: Limitations of Ferroptosis Inhibitors on the Doxorubicin-Induced Cardiotoxicity

doi: 10.3390/antiox15010027

Figure Lengend Snippet: Ferroptosis inhibitors do not alleviate doxorubicin-induced arrhythmic changes in mice. ( A ) Electrocardiogram (ECG) traces recorded on day 1 after DOX administration with or without ferroptosis inhibitors, showing various arrhythmic events, including tachyarrhythmia, frequent premature ventricular contractions, irregular rhythm, and cardiac arrest. Red arrows indicate representative arrhythmic events, including premature ventricular contractions and irregular R−R intervals. ( B ) ECG traces recorded on day 21, demonstrating the progression of arrhythmic changes, including atrioventricular (AV) block and bradycardia with AV block, indicated by red arrows. n = 4 per group. Data are presented as mean ± S.D., where n represents the number of individual mice per group. Statistical significance was determined by one-way ANOVA followed by Sidak’s multiple comparison test.

Article Snippet: ECG recordings were conducted for 22 days, with each mouse housed individually in a cage equipped with a receiver (RPC-1, Data Sciences International).

Techniques: Blocking Assay, Comparison