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Bnip3 mediates doxorubicin-induced cardiomyocyte pyroptosis via caspase-3/GSDME
Xinbin Zheng1, Ting Zhong1, Yeshuo Ma1
1Xiangya School of Pharmaceutical Sciences, Central South University, Changsha, Hunan 41008, China.
Aims:
This study was aimed to investigate the role of GSDME-mediated pyroptosis in cardiac injury induced by Doxorubicin (DOX), and to evaluate the role of BH3-only protein Bcl-2/adenovirus E1B 19-kDa-interacting protein 3 (Bnip3) in regulation of DOX-induced pyroptosis.
Main Methods:
HL-1 cardiomyocytes and C57BL/6J mice were treated by DOX to establish DOX-induced cardiotoxicity in vitro and in vivo models, respectively. Cell transfection was applied to regulate the expression of caspase-3, GSDME and Bnip3. Western blot was used for measuring expression of protein level. LDH-cytotoxicity assay was used to detect the LDH release. The Flow cytometry analysis was used to detect the cell death. Echocardiography was used to determine the cardiac function. HE staining was used for observing pathological feature of heart tissues.
Key Findings:
Our results showed that GSDME-mediated pyroptosis was involved in DOX-induced cardiotoxicity in vivo. We showed that HL-1 cardiomyocytes exposed to DOX exhibited morphological features of pyroptosis in vitro. We also showed that DOX induced activation of caspase-3 and eventually triggered GSDME-dependent pyroptosis, which was reduced by the silence or inhibitor of caspase-3. We further showed that knockdown of GSDME inhibited DOX-induced cardiomyocyte pyroptosis in vitro. Finally, DOX increased the expression of Bnip3, whereas silencing of Bnip3 blunted cardiomyocyte pyroptosis induced by DOX, which was regulated through caspase-3 activation and GSDME cleavage.
Significance:
Our findings revealed a novel pathway that cardiomyocyte pyroptosis is regulated through Bnip3-caspase-3-GSDME pathway following DOX treatment, suggesting that Bnip3-dependent pyroptosis may offer a novel therapeutic strategy to reduce cardiotoxicity induced by DOX.
Insights
Doxorubicin (DOX) induces heart injury via GSDME-mediated pyroptosis. Silencing Bnip3 protein reduces this DOX-induced pyroptosis, offering a potential therapeutic strategy for cardiotoxicity.
Area of Science:
- Cardiovascular Research
- Cell Death Mechanisms
- Molecular Cardiology
Background:
- Doxorubicin (DOX) is a widely used chemotherapy agent with known cardiotoxicity.
- Pyroptosis, a programmed form of inflammatory cell death, has emerged as a significant contributor to various pathologies, including cardiac injury.
- The specific role of Gasdermin E (GSDME)-mediated pyroptosis in DOX-induced cardiotoxicity requires further elucidation.
Purpose of the Study:
- To investigate the role of GSDME-mediated pyroptosis in Doxorubicin (DOX)-induced cardiac injury.
- To evaluate the regulatory function of the BH3-only protein Bcl-2/adenovirus E1B 19-kDa-interacting protein 3 (Bnip3) in DOX-induced pyroptosis.
Main Methods:
- Established in vitro (HL-1 cardiomyocytes) and in vivo (C57BL/6J mice) models of DOX-induced cardiotoxicity.
- Utilized cell transfection to modulate the expression of caspase-3, GSDME, and Bnip3.
- Employed Western blot, LDH-cytotoxicity assay, flow cytometry, echocardiography, and HE staining to assess protein levels, cell death, cardiac function, and tissue pathology.
Main Results:
- GSDME-mediated pyroptosis was confirmed to be involved in DOX-induced cardiotoxicity in vivo and in vitro.
- DOX treatment led to caspase-3 activation and subsequent GSDME-dependent pyroptosis, which was mitigated by caspase-3 inhibition.
- Knockdown of GSDME significantly reduced DOX-induced cardiomyocyte pyroptosis.
- DOX increased Bnip3 expression; silencing Bnip3 attenuated DOX-induced pyroptosis via caspase-3 activation and GSDME cleavage.
Conclusions:
- A novel Bnip3-caspase-3-GSDME pathway regulating cardiomyocyte pyroptosis following DOX treatment was identified.
- Bnip3-dependent pyroptosis presents a potential therapeutic target for mitigating Doxorubicin-induced cardiotoxicity.
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