Male Sprague-Dawley rats (6-8 weeks old; weighed from 210 to 230 g) were purchased from HFK Bioscience Co. Ltd. Rats were randomly assigned to four groups (n = 6 per group). The first was the control group, which were treated daily with 0.5 ml of 0.9% saline by intraperitoneal injection for 14 days, and there were three DOX model groups, which were treated three times weekly with 2.5 mg/kg of DOX by intraperitoneal injection for 14 weeks. At day 14, mice in the DOX model groups were infected through an intramyocardial injection of either control shNC or shMettl14 (1 x 10⁹ titer) at three distinct locations in the left ventricular free wall three times a week for 2 weeks, and they were treated daily with 30 mg/kg of ferroptosis inducer erastin (MedChemExpress, USA) through intragastric administration or vehicle control (Saline) for 2 weeks.

Male Sprague-Dawley rats (6-8 weeks old; weighed from 210 to 230 g) were purchased from HFK Bioscience Co. Ltd. Rats were randomly assigned to four groups (n = 6 per group). The first was the control group, which were treated daily with 0.5 ml of 0.9% saline by intraperitoneal injection for 14 days, and there were three DOX model groups, which were treated three times weekly with 2.5 mg/kg of DOX by intraperitoneal injection for 14 weeks. At day 14, mice in the DOX model groups were infected through an intramyocardial injection of either control shNC or shMettl14 (1 x 10⁹ titer) at three distinct locations in the left ventricular free wall three times a week for 2 weeks, and they were treated daily with 30 mg/kg of ferroptosis inducer erastin (MedChemExpress, USA) through intragastric administration or vehicle control (Saline) for 2 weeks.

Male Sprague-Dawley rats (6-8 weeks old; weighed from 210 to 230 g) were purchased from HFK Bioscience Co. Ltd. Rats were randomly assigned to four groups (n = 6 per group). The first was the control group, which were treated daily with 0.5 ml of 0.9% saline by intraperitoneal injection for 14 days, and there were three DOX model groups, which were treated three times weekly with 2.5 mg/kg of DOX by intraperitoneal injection for 14 weeks. At day 14, mice in the DOX model groups were infected through an intramyocardial injection of either control shNC or shMettl14 (1 x 10⁹ titer) at three distinct locations in the left ventricular free wall three times a week for 2 weeks, and they were treated daily with 30 mg/kg of ferroptosis inducer erastin (MedChemExpress, USA) through intragastric administration or vehicle control (Saline) for 2 weeks.

Male C57BL/6J mice aged 6-8 weeks with weights of 18-20 g were obtained from the Slack Laboratory Animal Co., Ltd. (Shanghai, China). Mice were allowed to acclimatize in the laboratory environment for 1 week before the beginning of the experiment. DCM model establishment: The mice were given a single intraperitoneal injection of 150 mg/kg 1% streptozotocin (STZ, V900890, Sigma, USA, dissolved in 0.1 mol/L sodium citrate buffer, pH = 4.4 - 4.6). Mouse blood from the tail vein was collected in each group of the model mice and tested by glucose meter (Accu-Chek Performa test strips, Roche, Accu-Chek Performa Combo, Roche, USA) on day 3, 5 and 7 after injection.

Male Sprague-Dawley rats (6-8 weeks old; weighed from 210 to 230 g) were purchased from HFK Bioscience Co. Ltd. Rats were randomly assigned to four groups (n = 6 per group). The first was the control group, which were treated daily with 0.5 ml of 0.9% saline by intraperitoneal injection for 14 days, and there were three DOX model groups, which were treated three times weekly with 2.5 mg/kg of DOX by intraperitoneal injection for 14 weeks. At day 14, mice in the DOX model groups were infected through an intramyocardial injection of either control shNC or shMettl14 (1 x 10⁹ titer) at three distinct locations in the left ventricular free wall three times a week for 2 weeks, and they were treated daily with 30 mg/kg of ferroptosis inducer erastin (MedChemExpress, USA) through intragastric administration or vehicle control (Saline) for 2 weeks.

Male Sprague-Dawley rats (6-8 weeks old; weighed from 210 to 230 g) were purchased from HFK Bioscience Co. Ltd. Rats were randomly assigned to four groups (n = 6 per group). The first was the control group, which were treated daily with 0.5 ml of 0.9% saline by intraperitoneal injection for 14 days, and there were three DOX model groups, which were treated three times weekly with 2.5 mg/kg of DOX by intraperitoneal injection for 14 weeks. At day 14, mice in the DOX model groups were infected through an intramyocardial injection of either control shNC or shMettl14 (1 x 10⁹ titer) at three distinct locations in the left ventricular free wall three times a week for 2 weeks, and they were treated daily with 30 mg/kg of ferroptosis inducer erastin (MedChemExpress, USA) through intragastric administration or vehicle control (Saline) for 2 weeks.

Adult male C57BL/6J mice weighing 20 ± 2 g were purchased from Chongqing Tengxin Biotechnology. Mice were housed at 22 with a 12 h light/dark cycle with free access to food and water. The cardiotoxicity mice model was induced by intraperitoneal injection of 5-FU (30 mg/kg) for 7 days. The cardiotoxicity mice were randomly divided into five groups: model group (normal saline), Res low, medium, high dose group (1, 2, 4 mg/kg) and Fer-1 positive control group (2.5 mg/kg). These mice were given Res or Fer-1 once a day for 3 weeks, with the body weight being recorded. Then, the mice were euthanized, blood samples and heart tissue were collected.

All animal experiment protocols were implemented in accordance with the National Institutes of Health (NIH) guidelines, and the procedures were approved by the Animal Ethics Committee of Southwest University. C57BL/6J male mice, 8-10 weeks old, weighing 20 ± 2 g, were used in this study. Mice were housed under standard conditions at 22-24 with a 12 h light/12 h darkness cycle and free access to food and tap water. Thirty-six mice were randomly divided into six groups: control (N = 8), IMA group (50 mg/kg) (N = 8), Low-Ber (20 mg/kg) + IMA group (N = 8), Medium-Ber (40 mg kg1) + IMA group (N = 8), High-Ber (80 mg/kg) + IMA group (N = 8), and Fer-1 (1 mg/kg) + IMA group (N = 8). IMA was given intraperitoneally for 14 days. Ber was given orally 2 h before IMA treatment and Fer-1 was given intraperitoneally 2 h before IMA treatment.

Male C57BL/6J mice aged 6-8 weeks with weights of 18-20 g were obtained from the Slack Laboratory Animal Co., Ltd. (Shanghai, China). Mice were allowed to acclimatize in the laboratory environment for 1 week before the beginning of the experiment. DCM model establishment: The mice were given a single intraperitoneal injection of 150 mg/kg 1% streptozotocin (STZ, V900890, Sigma, USA, dissolved in 0.1 mol/L sodium citrate buffer, pH = 4.4 - 4.6). Mouse blood from the tail vein was collected in each group of the model mice and tested by glucose meter (Accu-Chek Performa test strips, Roche, Accu-Chek Performa Combo, Roche, USA) on day 3, 5 and 7 after injection.

Two-month-old male New Zealand rabbits purchased from the Medical Experimental Animal Center of Bengbu Medical College were used as experimental subjects. Streptozotocin was dissolved in sterile saline and intraperitoneally injected into the rabbits at a dose of 80 mg/kg. The rabbits were allowed to eat freely after receiving the injection. The fasting blood glucose levels of the rabbits were monitored regularly. The diabetic rabbit model was considered successfully established when the fasting blood glucose level was measured as 11 mmol/L twice or 14 mmol/L once. Following successful modelling, grouping was performed as follows: blank control group (Con-Group), diabetic rabbit group (DM-Group), diabetic rabbit + every other day curcumin administration group (Qod-Group), and diabetic rabbit + daily administration group (Qd-Group).

Male C57BL/6J mice were housed in a temperature- and humidity-controlled room, fed a commercial diet (CRF-1; Oriental Yeast Co. Ltd.), and given free access to water. GPx4 Tg mice and GPx4 hetKO mice were produced as previously described. In these gene-manipulated mice, GPx4 was systemically overexpressed or absent, respectively. These strains were backcrossed with C57BL/6J mice in our laboratory. The DIC model was reproduced as previously reported, with some modification. Briefly, DOX (6 mg/kg, body weight) was administered to mice via tail vein at days 0, 2, and 4.

DOX (D1515, Sigma-Aldrich, St Louis, MO, USA; 6 mg/kg, dissolved in distilled water) was administrated to C57BL/6J Male mice (8-10 weeks old, 21-24 g) via the tail vein on days 0, 2, and 4.Ethoxyquin(E0237, Tokyo Chemical Industry, Tokyo, Japan; 100 umol/kg, once a day) was orally administrated every day from days 0 to 14. Ethoxyquinwas dissolved in polyethylene glycol (PEG; 28214-05, Nacalai Tesque, Kyoto, Japan,ethoxyquin10 uL in 990 uL PEG), and the solution was then diluted in the same amount of normal saline (873311, Otsuka Pharmaceutical, Tokyo, Japan).

The set of animal experiments was designed to evaluate the effectiveness of liquiritin on doxorubicin-induced cardiotoxicity as were as ferroptosis were explored. Mice were randomly divided into 5 groups: (1) the control group; (2) doxorubicin group; (3) the doxorubicin plus liquiritin group (20 mg/kg); (4) the doxorubicin plus liquiritin group (40 mg/kg); (5) the doxorubicin plus liquiritin group (80 mg/kg) (Han et al.2022; Mou et al.2021). The control group and doxorubicin group were given equal volume of 0.5% sodium carboxymethylcellulose; the doxorubicin plus liquiritin groups were given different doses of liquiritin (0.5%) sodium carboxymethylcellulose co-suspension) by intragastric administration 7 days in advance once a daily. On day 8, groups (2), (3), (4), and (5) were given a single intraperitoneal injection of 15 mg/kg of DOX to establish a model of doxorubicin-induced cardiotoxicity; and group (1) was given an equal volume of saline intraperitoneally.

To simulate the animal model of diabetic cardiomyopathy, male db/+ mice and db/db mice (age, 7 weeks, weight, 24 g) were fed a normal diet for 4 weeks and kept at 24 under a 14-h light/8-h dark cycle. The animals were purchased from the Model Animal Research Center of Nanjing University (Nanjing, China). Diabetic mice were intracoronarily administered equal volumes (80 ul) of adenoviruses Ad-ZFAS1, Ad-sh-ZFAS1, Ad-CCND2, Ad-sh-CCND2 or Ad-NC.33 miR-150-5p mimics and mimic control (NC) were injected into the tail vein of mice (50 ug/kg) every 15 days for 12 weeks. Db/db mice were treated with or without ferrostatin-1 (Fer-1, ferroptosis inhibitor; Sigma-Aldrich, 5 mg/kg) for an additional 12 weeks.

To simulate the animal model of diabetic cardiomyopathy, male db/+ mice and db/db mice (age, 7 weeks, weight, 24 g) were fed a normal diet for 4 weeks and kept at 24 under a 14-h light/8-h dark cycle. The animals were purchased from the Model Animal Research Center of Nanjing University (Nanjing, China). Diabetic mice were intracoronarily administered equal volumes (80 ul) of adenoviruses Ad-ZFAS1, Ad-sh-ZFAS1, Ad-CCND2, Ad-sh-CCND2 or Ad-NC.33 miR-150-5p mimics and mimic control (NC) were injected into the tail vein of mice (50 ug/kg) every 15 days for 12 weeks. Db/db mice were treated with or without ferrostatin-1 (Fer-1, ferroptosis inhibitor; Sigma-Aldrich, 5 mg/kg) for an additional 12 weeks.

To simulate the animal model of diabetic cardiomyopathy, male db/+ mice and db/db mice (age, 7 weeks, weight, 24 g) were fed a normal diet for 4 weeks and kept at 24 under a 14-h light/8-h dark cycle. The animals were purchased from the Model Animal Research Center of Nanjing University (Nanjing, China). Diabetic mice were intracoronarily administered equal volumes (80 ul) of adenoviruses Ad-ZFAS1, Ad-sh-ZFAS1, Ad-CCND2, Ad-sh-CCND2 or Ad-NC.33 miR-150-5p mimics and mimic control (NC) were injected into the tail vein of mice (50 ug/kg) every 15 days for 12 weeks. Db/db mice were treated with or without ferrostatin-1 (Fer-1, ferroptosis inhibitor; Sigma-Aldrich, 5 mg/kg) for an additional 12 weeks.

In all experiments using KO mice (male, 8-12 weeks), we used age-matched, tamoxifen-treated MerCreMer single-genotype animals as controls. For inducing cardiac-specific knockout of MITOL, an ethanol-corn oil emulsion of tamoxifen (TAX; H5648, Sigma-Aldrich) was injected intraperitoneally per day of 30 mg tamoxifen/kg body weight (150 mg/kg total) for 5 days.

A total of 24 SD male rats weighing 200-210 g from the Vital River Laboratory Animal Technology Co., Ltd. were divided randomly into control, ADR, ADR+AsIV, and AsIV group (n = 6). AsIV was administered by gavage at a dose of 10 mg/kg/day over a period of five weeks. ADR was administered intraperitoneally once a week (30 mg/kg/week) for five weeks. Controls were administered saline intraperitoneally (i.p.) at the same dose as ADR and intragastrically at the same dose as AsIV.

All animal experiment protocols were implemented in accordance with the National Institutes of Health (NIH) guidelines, and the procedures were approved by the Animal Ethics Committee of Southwest University. C57BL/6J male mice, 8-10 weeks old, weighing 20 ± 2 g, were used in this study. Mice were housed under standard conditions at 22-24 with a 12 h light/12 h darkness cycle and free access to food and tap water. Thirty-six mice were randomly divided into six groups: control (N = 8), IMA group (50 mg/kg) (N = 8), Low-Ber (20 mg/kg) + IMA group (N = 8), Medium-Ber (40 mg kg1) + IMA group (N = 8), High-Ber (80 mg/kg) + IMA group (N = 8), and Fer-1 (1 mg/kg) + IMA group (N = 8). IMA was given intraperitoneally for 14 days. Ber was given orally 2 h before IMA treatment and Fer-1 was given intraperitoneally 2 h before IMA treatment.

Two-month-old male New Zealand rabbits purchased from the Medical Experimental Animal Center of Bengbu Medical College were used as experimental subjects. Streptozotocin was dissolved in sterile saline and intraperitoneally injected into the rabbits at a dose of 80 mg/kg. The rabbits were allowed to eat freely after receiving the injection. The fasting blood glucose levels of the rabbits were monitored regularly. The diabetic rabbit model was considered successfully established when the fasting blood glucose level was measured as 11 mmol/L twice or 14 mmol/L once. Following successful modelling, grouping was performed as follows: blank control group (Con-Group), diabetic rabbit group (DM-Group), diabetic rabbit + every other day curcumin administration group (Qod-Group), and diabetic rabbit + daily administration group (Qd-Group).

Male C57BL/6J mice aged 6-8 weeks with weights of 18-20 g were obtained from the Slack Laboratory Animal Co., Ltd. (Shanghai, China). Mice were allowed to acclimatize in the laboratory environment for 1 week before the beginning of the experiment. DCM model establishment: The mice were given a single intraperitoneal injection of 150 mg/kg 1% streptozotocin (STZ, V900890, Sigma, USA, dissolved in 0.1 mol/L sodium citrate buffer, pH = 4.4 - 4.6). Mouse blood from the tail vein was collected in each group of the model mice and tested by glucose meter (Accu-Chek Performa test strips, Roche, Accu-Chek Performa Combo, Roche, USA) on day 3, 5 and 7 after injection.

The set of animal experiments was designed to evaluate the effectiveness of liquiritin on doxorubicin-induced cardiotoxicity as were as ferroptosis were explored. Mice were randomly divided into 5 groups: (1) the control group; (2) doxorubicin group; (3) the doxorubicin plus liquiritin group (20 mg/kg); (4) the doxorubicin plus liquiritin group (40 mg/kg); (5) the doxorubicin plus liquiritin group (80 mg/kg) (Han et al.2022; Mou et al.2021). The control group and doxorubicin group were given equal volume of 0.5% sodium carboxymethylcellulose; the doxorubicin plus liquiritin groups were given different doses of liquiritin (0.5%) sodium carboxymethylcellulose co-suspension) by intragastric administration 7 days in advance once a daily. On day 8, groups (2), (3), (4), and (5) were given a single intraperitoneal injection of 15 mg/kg of DOX to establish a model of doxorubicin-induced cardiotoxicity; and group (1) was given an equal volume of saline intraperitoneally.

All animal protocols were approved by the Animal Care and Use Committee of TaizhouHospital, affiliated to Zhejiang University (Taizhou, China). Twenty-four 2-month-old male Wistar rats weighing 190-220 g were purchased from the Experimental Animal Center of Basi Medicine, Zhejiang Chinese Medical University. The animals were reared under a 12 h light/12 h dark cycle at a relative humidity of 55 ± 5% and temperature of 23 ± 2 , with unrestricted access to food and water. All animals were acclimatized to laboratory conditions for 1 week before the experiments and were randomly divided into four groups: control group (CG, n = 6); LCZ696 group (LCZ, n = 6); DOX group (DOX, n = 6); and DOX + LCZ696 group (DOX + LCZ, n = 6). The CG received saline solution by gavage for 6 weeks (2 mL/day), while the treatment groups received DOX (Cat. HY-15142A, MedChemExpress, USA), LCZ696 (Cat. HY-18204A, MedChemExpress, USA), or DOX + LCZ696. DOX was administered at a dose of 2.5 mg/kg once a week for 6 weeks via tailvein injection. LCZ696 (60 mg/kg/day) was administered by gavage for 6 weeks. Body weight was measured weekly.

All animal protocols were approved by the Animal Care and Use Committee of TaizhouHospital, affiliated to Zhejiang University (Taizhou, China). Twenty-four 2-month-old male Wistar rats weighing 190-220 g were purchased from the Experimental Animal Center of Basi Medicine, Zhejiang Chinese Medical University. The animals were reared under a 12 h light/12 h dark cycle at a relative humidity of 55 ± 5% and temperature of 23 ± 2 , with unrestricted access to food and water. All animals were acclimatized to laboratory conditions for 1 week before the experiments and were randomly divided into four groups: control group (CG, n = 6); LCZ696 group (LCZ, n = 6); DOX group (DOX, n = 6); and DOX + LCZ696 group (DOX + LCZ, n = 6). The CG received saline solution by gavage for 6 weeks (2 mL/day), while the treatment groups received DOX (Cat. HY-15142A, MedChemExpress, USA), LCZ696 (Cat. HY-18204A, MedChemExpress, USA), or DOX + LCZ696. DOX was administered at a dose of 2.5 mg/kg once a week for 6 weeks via tailvein injection. LCZ696 (60 mg/kg/day) was administered by gavage for 6 weeks. Body weight was measured weekly.

Male Sprague-Dawley (SD) rats (160-180 g) were purchased from the Experimental Animal Center of Guangzhou University of Chinese Medicine. After 1 week of acclimatization, the rats were randomly divided into two groups: the control group (standard diet,n = 6) and the high-fat diet (HFD) group. The control group was given a standard diet for 8 weeks, and the HFD group was given a high-fat diet (feed item No. D12451, Guangdong Medical Laboratory Animal Center) for 8 weeks. Subsequently, intraperitoneal injection of streptozotocin (STZ, 40 mg/kg) was performed in the HFD group for inducing diabetic symptoms, and an equal volume of saline was given to the control group. A diabetes model was successfully established under the condition that the fasting blood glucose levels were > 16.7 mmol/L over three consecutive days. Diabetic rats in the HFD group were divided into five groups: a DCM group, three AS-IV treatment groups (20, 40, and 80 mg/kg/day by gavage,n = 6), and an atorvastatin treatment group (ALE group, used as a positive control drug, 10 mg/kg, by gavage,n = 6). Rats were given the standard diet during drug gavage. After 12 weeks of treatment, all animals were anesthetized and then euthanized by intraperitoneal injection with an overdose of pentobarbital sodium. The animal experiments in this study were randomized and single-blind.

A total of 18 Wistar rats (250~300 g) were purchased from Hunan slake Jingda experimental animal Co., Ltd. The rats were randomly divided into the Sham group, I/R group, and I/R + ATV group (n = 6/group).They received standard diet and water before myocardial I/R. Rats in the I/R + ATV group were orally treated with ATV (10 mg/kg/d) for 2 weeks before myocardial I/R (9).The Sham and I/R model rats were constructed as follows: The rats were anesthetized with sodium pentobarbital (50 mg/kg, intraperitoneal injection), ligation of the left anterior descending branch with 4-0 silk thread for 30 min, and then reperfusion for 180 min. In the sham control group, the entire procedure was performed with silk thread passing below the coronary artery, but the LAD coronary artery was not ligated. At the end of reperfusion, the rats were given excessive isoflurane for 10 min and sacrificed by bloodletting. Then the rat myocardial tissues were isolated for subsequent detection.

All animal experiment protocols were implemented in accordance with the National Institutes of Health (NIH) guidelines, and the procedures were approved by the Animal Ethics Committee of Southwest University. C57BL/6J male mice, 8-10 weeks old, weighing 20 ± 2 g, were used in this study. Mice were housed under standard conditions at 22-24 with a 12 h light/12 h darkness cycle and free access to food and tap water. Thirty-six mice were randomly divided into six groups: control (N = 8), IMA group (50 mg/kg) (N = 8), Low-Ber (20 mg/kg) + IMA group (N = 8), Medium-Ber (40 mg kg1) + IMA group (N = 8), High-Ber (80 mg/kg) + IMA group (N = 8), and Fer-1 (1 mg/kg) + IMA group (N = 8). IMA was given intraperitoneally for 14 days. Ber was given orally 2 h before IMA treatment and Fer-1 was given intraperitoneally 2 h before IMA treatment.

Male Wistar rats (250-300 g,n = 230) were purchased from Vitalriver. Ten rats were recruited in each group in the experiment. The rats in the control, DOX, FER-1 + DOX, NEC-1 + DOX, 3-MA + DOX, and Emricasan + DOX groups were used for the overall survival analysis, and those from control and DOX groups were used for detection of the expression of PTGS2 in the indicated organs of rats.

C57BL/6 male mice (20-25 g) at 7 weeks were purchased from the Vital River Laboratory Animal Technology Co., Ltd. Two doses of DOX was administrated by intraperitoneal injection, 15 mg/kg at Day 1, and 10 mg/kg at Day 8. Mice were then killed at Day 15 after transthoracic echocardiography examination.

Male Sprague-Dawley rats (6-8 weeks old; weighed from 210 to 230 g) were purchased from HFK Bioscience Co. Ltd. Rats were randomly assigned to four groups (n = 6 per group). The first was the control group, which were treated daily with 0.5 ml of 0.9% saline by intraperitoneal injection for 14 days, and there were three DOX model groups, which were treated three times weekly with 2.5 mg/kg of DOX by intraperitoneal injection for 14 weeks. At day 14, mice in the DOX model groups were infected through an intramyocardial injection of either control shNC or shMettl14 (1 x 10⁹ titer) at three distinct locations in the left ventricular free wall three times a week for 2 weeks, and they were treated daily with 30 mg/kg of ferroptosis inducer erastin (MedChemExpress, USA) through intragastric administration or vehicle control (Saline) for 2 weeks.

Mice were reseparated into 5 groups: control, Dox, Dox + Fer-1, Dox + EGCG, and Dox + EGCG+compound C groups. The Dox group mice were administered 6 intraperitoneal (ip) injections of 2.5 mg/kg Dox over 3 weeks for a cumulative dose of 15 mg/kg. Mice in the Dox + Fer-1 group were ip injected with 1 mg/kg/d Fer-1 for 2 weeks as in the Dox group. Mice in the Dox + EGCG group were intragastrically (ig) injected 20 mg/kg/d EGCG (dissolved in normal saline) for six consecutive weeks; Dox was administered 1 h prior to this as in the Dox group. Mice in the Dox + EGCG + compound C group were treated using the same method as in the Dox + EGCG group for four consecutive weeks, followed by ip injections of 10 mg/kg/d compound C for 2 weeks. Mice in the control groups were administered an equal volume of normal saline via gavage for 6 weeks.

Male C57/BL mice (aged between 6 and 8 weeks and weighing 20 ± 2 g) were randomly divided into 6 groups (n = 6 mice per group) with equal number of mice in each group. The groups included saline control group (control group): 200 ulxd-1 saline intraperitoneally administered to the mice for 10 days; DOX model group (DOX group): 200 ulxd-1 saline intraperitoneally administered to the mice for 10 days and a single intraperitoneal administration of 10 mgxkg-1 DOX (HY-15,142, MCE, China) to the mice on the seventh day.

All animal procedures were approved by the Animal Care and Use Committees at the Second Affiliated Hospital of Nanchang University (Nanchang, China) and Zhongshan Hospital Fudan University (Shanghai, China). In brief, adult male C57BL/6J mice were intraperitoneally delivered TZM at a dose of 10 mg/kg once per week for 6 weeks. A cohort of mice received Empagliflozin at a dose of 10 mg/kg twice per week for 6 weeks. All mice were maintained on a 12/12-light/dark cycle with free access to tap water and lab chow until experimentation. Blood glucose and serum triglyceride levels were obtained using a commercial glucometer and ELISA commercial kits, respectively. Serum levels oflactic dehydrogenase(LDH) and troponin I were measured using chemiluminescent immunoassays. To discern the involvement of ferroptosis in TZM-induced cardiotoxicity, a cohort of TZM challenged C57BL/6J mice (10 mg/kg once per week for 4 weeks) also received the ferroptosis inhibitor liproxtatin-1 (LIP-1, 10 mg/kg, i.p., every other day) during the entire duration of TZM challenge.

A total of 18 Wistar rats (250~300 g) were purchased from Hunan slake Jingda experimental animal Co., Ltd. The rats were randomly divided into the Sham group, I/R group, and I/R + ATV group (n = 6/group).They received standard diet and water before myocardial I/R. Rats in the I/R + ATV group were orally treated with ATV (10 mg/kg/d) for 2 weeks before myocardial I/R (9).The Sham and I/R model rats were constructed as follows: The rats were anesthetized with sodium pentobarbital (50 mg/kg, intraperitoneal injection), ligation of the left anterior descending branch with 4-0 silk thread for 30 min, and then reperfusion for 180 min. In the sham control group, the entire procedure was performed with silk thread passing below the coronary artery, but the LAD coronary artery was not ligated. At the end of reperfusion, the rats were given excessive isoflurane for 10 min and sacrificed by bloodletting. Then the rat myocardial tissues were isolated for subsequent detection.

All animal experiment protocols were implemented in accordance with the National Institutes of Health (NIH) guidelines, and the procedures were approved by the Animal Ethics Committee of Southwest University. C57BL/6J male mice, 8-10 weeks old, weighing 20 ± 2 g, were used in this study. Mice were housed under standard conditions at 22-24 with a 12 h light/12 h darkness cycle and free access to food and tap water. Thirty-six mice were randomly divided into six groups: control (N = 8), IMA group (50 mg/kg) (N = 8), Low-Ber (20 mg/kg) + IMA group (N = 8), Medium-Ber (40 mg kg1) + IMA group (N = 8), High-Ber (80 mg/kg) + IMA group (N = 8), and Fer-1 (1 mg/kg) + IMA group (N = 8). IMA was given intraperitoneally for 14 days. Ber was given orally 2 h before IMA treatment and Fer-1 was given intraperitoneally 2 h before IMA treatment.

Two-month-old male New Zealand rabbits purchased from the Medical Experimental Animal Center of Bengbu Medical College were used as experimental subjects. Streptozotocin was dissolved in sterile saline and intraperitoneally injected into the rabbits at a dose of 80 mg/kg. The rabbits were allowed to eat freely after receiving the injection. The fasting blood glucose levels of the rabbits were monitored regularly. The diabetic rabbit model was considered successfully established when the fasting blood glucose level was measured as 11 mmol/L twice or 14 mmol/L once. Following successful modelling, grouping was performed as follows: blank control group (Con-Group), diabetic rabbit group (DM-Group), diabetic rabbit + every other day curcumin administration group (Qod-Group), and diabetic rabbit + daily administration group (Qd-Group).

All animal experiment protocols were implemented in accordance with the National Institutes of Health (NIH) guidelines, and the procedures were approved by the Animal Ethics Committee of Southwest University. C57BL/6J male mice, 8-10 weeks old, weighing 20 ± 2 g, were used in this study. Mice were housed under standard conditions at 22-24 with a 12 h light/12 h darkness cycle and free access to food and tap water. Thirty-six mice were randomly divided into six groups: control (N = 8), IMA group (50 mg/kg) (N = 8), Low-Ber (20 mg/kg) + IMA group (N = 8), Medium-Ber (40 mg kg1) + IMA group (N = 8), High-Ber (80 mg/kg) + IMA group (N = 8), and Fer-1 (1 mg/kg) + IMA group (N = 8). IMA was given intraperitoneally for 14 days. Ber was given orally 2 h before IMA treatment and Fer-1 was given intraperitoneally 2 h before IMA treatment.

To simulate the animal model of diabetic cardiomyopathy, male db/+ mice and db/db mice (age, 7 weeks, weight, 24 g) were fed a normal diet for 4 weeks and kept at 24 under a 14-h light/8-h dark cycle. The animals were purchased from the Model Animal Research Center of Nanjing University (Nanjing, China). Diabetic mice were intracoronarily administered equal volumes (80 ul) of adenoviruses Ad-ZFAS1, Ad-sh-ZFAS1, Ad-CCND2, Ad-sh-CCND2 or Ad-NC.33 miR-150-5p mimics and mimic control (NC) were injected into the tail vein of mice (50 ug/kg) every 15 days for 12 weeks. Db/db mice were treated with or without ferrostatin-1 (Fer-1, ferroptosis inhibitor; Sigma-Aldrich, 5 mg/kg) for an additional 12 weeks.

To simulate the animal model of diabetic cardiomyopathy, male db/+ mice and db/db mice (age, 7 weeks, weight, 24 g) were fed a normal diet for 4 weeks and kept at 24 under a 14-h light/8-h dark cycle. The animals were purchased from the Model Animal Research Center of Nanjing University (Nanjing, China). Diabetic mice were intracoronarily administered equal volumes (80 ul) of adenoviruses Ad-ZFAS1, Ad-sh-ZFAS1, Ad-CCND2, Ad-sh-CCND2 or Ad-NC.33 miR-150-5p mimics and mimic control (NC) were injected into the tail vein of mice (50 ug/kg) every 15 days for 12 weeks. Db/db mice were treated with or without ferrostatin-1 (Fer-1, ferroptosis inhibitor; Sigma-Aldrich, 5 mg/kg) for an additional 12 weeks.

To simulate the animal model of diabetic cardiomyopathy, male db/+ mice and db/db mice (age, 7 weeks, weight, 24 g) were fed a normal diet for 4 weeks and kept at 24 under a 14-h light/8-h dark cycle. The animals were purchased from the Model Animal Research Center of Nanjing University (Nanjing, China). Diabetic mice were intracoronarily administered equal volumes (80 ul) of adenoviruses Ad-ZFAS1, Ad-sh-ZFAS1, Ad-CCND2, Ad-sh-CCND2 or Ad-NC.33 miR-150-5p mimics and mimic control (NC) were injected into the tail vein of mice (50 ug/kg) every 15 days for 12 weeks. Db/db mice were treated with or without ferrostatin-1 (Fer-1, ferroptosis inhibitor; Sigma-Aldrich, 5 mg/kg) for an additional 12 weeks.