Abstract
Doxorubicin (Dox), an effective therapy in different types of cancer, is known to exhibit cardiotoxic effects. Despite previous studies indicating the benefits of dapagliflozin (DAPA) in patients experiencing heart failure, it remains uncertain whether DAPA exerts a protective effect on Dox-induced cardiac dysfunction. Signal transducer and activator of transcription 3 (STAT3) participates in various mechanisms of cardioprotection. Herein, we aimed to investigate the effects of DAPA on Dox-induced cardiotoxicity and the role of STAT3. Sprague–Dawley rats were pretreated with oral DAPA for 6 weeks followed by Dox for 4 weeks. Sequential echocardiography was applied to assess cardiac function. For in vitro analysis, cardiomyocytes were treated with 10 μM DAPA and subsequently exposed to 1 μM Dox. The expression of reactive oxygen species- and apoptosis-related proteins was measured. Using STAT3 siRNA, we further examined the effects of STAT3 effect on DAPA-associated protection against Dox-induced apoptosis. In rats treated with Dox, DAPA significantly reduced cardiac fibrosis and improved cardiac function and hemodynamics. Additionally, DAPA effectively inhibited Dox-induced apoptosis and reactive oxygen species (ROS) in cardiomyocytes. Mechanistically, we showed that DAPA decreased cardiac expression of Bax and cleaved caspase 3 but increased Bcl-2 expression. DAPA also significantly rescued Dox-suppressed STAT3 expression. Conversely, knocking down STAT3 in cardiomyocytes reversed the DAPA-related protective effects on Dox-induced cell apoptosis and ROS. Collectively, our findings indicate that DAPA could be useful for preventing Dox-induced cardiotoxicity by restoring STAT3.
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The data are available upon the reasonable request to the corresponding author.
Abbreviations
- Dox:
-
Doxorubicin
- Dapa:
-
Dapagliflozin
- STZ:
-
Streptozotocin
- ER:
-
Endoplasmic reticulum
- ROS:
-
Reactive oxygen species
- SGLT2:
-
Sodium glucose cotransporter
- NHI:
-
National Health Insurance
- NHIRD:
-
National Health Insurance Research Database
- MACCEs:
-
Major adverse cardio- and cerebrovascular events
- CKD:
-
Chronic kidney disease
- ESRD:
-
End-stage renal disease
- COPD:
-
Chronic obstructive pulmonary disease
- IVSd:
-
Interventricular septum thickness in diastole
- LVIDd:
-
Left ventricular internal diameter; in diastole
- EF:
-
Ejection fraction
- FS:
-
Fractional shortening
- P-V:
-
Pressure–volume
- HE:
-
Hematoxylin-eosin
- Ves:
-
End-systolic
- Ved:
-
Diastolic volumes
- Pes:
-
End-systolic
- Ped:
-
Diastolic pressures
- +dP/dt and -dP/dt:
-
Maximal velocity of pressure rise and fall
- Ea:
-
Arterial elastance
- tau:
-
Time constant of isovolumic pressure decay
- ESPVR:
-
End-systolic pressure–volume relationship
- EDPVR:
-
End-diastolic pressure–volume relationship
- IVC:
-
Inferior vena cava
- MTT:
-
3-(4,5-Dimethyl-2-thiazolyl)-2,5-dimethyl-2H-tetrazolium bromide
- H2DCF‑DA:
-
Fluorescent 2′,7′‑dichlorofluorescindiacetate
- PI:
-
Propidium iodide
- TUNEL:
-
Terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling
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Acknowledgements
We especially thank the support from Professor Wei-Jan Chen in Linkou Chang Gung Memorial Hospital/Chang Gung University, Taiwan
Funding
This study was supported by Chi-Mei Medical Center, Ministry of Science and Technology (MOST105-2628-B-384-001-MY3; 108-2628-B-384), National Health Research Institute (NHRI-EX106-10618SC).
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All authors were involved in the conception and design of the study and data interpretation. WC and YL drafted the paper and performed data analysis. WC and YL were involved in the data analysis and interpretation. All authors critically revised the paper and approved it for submission.
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This study was approved by the Institutional Review Board of Chi-Mei Medical Center (CV code: 10406-E01). Given that the data is derived from the NHIRD databank, the consent to participate is not applicable.
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204_2022_3298_MOESM2_ESM.tif
Supplementary file2 Supplement Figure 2. The changes in (A) body weight (BW), (B) blood sugar (BS), and (C) heart rate (HR) in the control, dapagliflozin (DAPA), doxorubicin (Dox) and Dox + DAPA groups (N=6–8) (TIF 740 KB)
204_2022_3298_MOESM3_ESM.tif
Supplementary file3 Supplement Figure 3. The effects of dapagliflozin (DAPA) on survival, cardiac morphology and lung injury in rats treated with doxorubicin (Dox). (A) The survival curve, (B) harvested hearts, (C) heart-to-body weight ratio, (D) heart-to-tibial bone length ratio, and (E) wet-dry lung ratio in the control, DAPA, Dox, and Dox+DAPA groups. *P < 0.05, **P < 0.01, and ***P < 0.001 compared with the indicated groups (N=6–8) (TIF 2282 KB)
204_2022_3298_MOESM4_ESM.tif
Supplementary file4 Supplement Figure 4. The effects of dapagliflozin (DAPA) on STAT3 in cardiomyocytes of rat after doxorubicin (Dox) treatment. (A) Representative sections of hearts stained with STAT3 (green) and F-actin (blue); scale bars, 20 µm. (B) Quantification of STAT3 expression in the indicated groups of rats. *P < 0.05, and ***P < 0.001 compared with the indicated groups (N=5) (TIF 12685 KB)
204_2022_3298_MOESM5_ESM.tif
Supplementary file5 Supplement Figure 5. The efficiency of transfection in cardiomyocytes treated with STAT3 siRNA compared with cells transfected with control siRNA. Abbreviations: Dox: doxorubicin; Dapa: dapagliflozin; STZ: Streptozotocin; ER: endoplasmic reticulum; ROS: reactive oxygen species; SGLT2: sodium glucose cotransporter; NHI: National Health Insurance; NHIRD: National Health Insurance Research Database; MACCEs: major adverse cardio- and cerebrovascular events; CKD: chronic kidney disease; ESRD: end-stage renal disease; COPD: chronic obstructive pulmonary disease; IVSd: interventricular septum thickness in diastole; LVIDd: left ventricular internal diameter; in diastole; EF: ejection fraction; FS: fractional shortening; P-V: Pressure–volume; HE: hematoxylin–eosin; Ves: end-systolic; Ved: diastolic volumes; Pes: end-systolic; Ped: diastolic pressures; +dP/dt and -dP/dt: maximal velocity of pressure rise and fall; Ea: arterial elastance; tau: time constant of isovolumic pressure decay; ESPVR: end-systolic pressure-volume relationship; EDPVR: end-diastolic pressure–volume relationship; IVC: inferior vena cava; MTT:3-(4,5-dimethyl-2-thiazolyl)-2,5-dimethyl-2H-tetrazolium bromide; H2DCF‑DA: fluorescent 2',7'‑dichlorofluorescindiacetate; PI: propidium iodide; TUNEL: terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling (TIF 422 KB)
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Chang, WT., Shih, JY., Lin, YW. et al. Dapagliflozin protects against doxorubicin-induced cardiotoxicity by restoring STAT3. Arch Toxicol 96, 2021–2032 (2022). https://doi.org/10.1007/s00204-022-03298-y
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DOI: https://doi.org/10.1007/s00204-022-03298-y