Beclin1 controls caspase-4 inflammsome activation and pyroptosis in mouse myocardial reperfusion-induced microvascular injury.

Abstract

BACKGROUND:

Myocardial reperfusion injury is often accompanied by cell death and inflammatory reactions. Recently, pyroptosis is gradually recognized as pivotal role in cardiovascular disease. However, little is known about the regulatory role of beclin1 in the control of caspase-4 activation and pyroptosis. The present study confirmed whether beclin1 regulates caspase-4 mediated pyroptosis and thereby protects Human Cardiac microvascular endothelial cells (HCMECs) against injury.

METHODS:

TTC and Evan's blue dye, western blot, immunofluorescence and immunohistochemistry staining were performed in wild mice and transgenic mice with overexpression of beclin 1(BECN1-Tg). CMECs were transfected with a beclin1 lentivirus. The cell cytotoxicity was analyzed by LDH-Cytotoxicity Assay Kit. The protein levels of autophagy protein (Beclin1, p62 and LC3II/LC3I) and caspase-4/GSDMD pathway were determined by western blot. Autophagic vacuoles in cells were monitored with RFP-GFP-LC3 using fluorescence microscope.

RESULTS:

I/R caused caspase-4 activity and gasdermin D expression increase in vivo and in vitro. Overexpression of beclin1 in heart tissue and CMECs suppressed the caspase-4 activity and decreased the levels of gasdermin D; meanwhile beclin1 overexpression also reduced IL-1β levels, promoted autophagy (p62 expression was inhibited while LC3II expression was increased) in the heart and CMECs. Interestingly, beclin1 overexpression increased animal survival and attenuated myocardial infarct size (45 ± 6.13 vs 22 ± 4.37), no-reflow area (39 ± 5.22 vs 16 ± 2.54) post-myocardial ischemia reperfusion.

CONCLUSIONS:

Induction of beclin-1 signaling can be a potential therapeutic target in myocardial reperfusion-induced microvascular injury. Video Abstract.

• Authors: Wenjing Sun, Hongquan Lu, Shujuan Dong, Rui Li, Yingjie Chu, Nan Wang, Yu Zhao, Yabin Zhang, Limeiting Wang, Lin Sun, Di Lu
• Journal: Cell communication and signaling : CCS (Cell Commun Signal) Vol. 19 Issue 1 Pg. 107 (11 03 2021) ISSN: 1478-811X [Electronic] England
• PMID: 34732218 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)
• Copyright: © 2021. The Author(s).

Chemical References

• Beclin-1
• Inflammasomes
• Map1lc3b protein, mouse
• Microtubule-Associated Proteins
• P62 protein, human
• RNA-Binding Proteins
• CASP4 protein, human
• Casp4 protein, mouse
• Caspases, Initiator

Topics

• Animals
• Autophagy (genetics)
• Beclin-1 (genetics)
• Caspases, Initiator (genetics)
• Disease Models, Animal
• Endothelial Cells (metabolism, pathology)
• Gene Expression Regulation (genetics)
• Humans
• Inflammasomes (genetics, metabolism)
• Mice
• Mice, Transgenic
• Microtubule-Associated Proteins (genetics)
• Microvessels (injuries, metabolism, pathology)
• Myocardial Infarction (genetics, pathology)
• Myocardial Reperfusion Injury (genetics, pathology)
• Myocytes, Cardiac (metabolism, pathology)
• Pyroptosis (genetics)
• RNA-Binding Proteins (genetics)