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.

Endothelial Autophagy in Coronary Microvascular Dysfunction and Cardiovascular Disease

“…Inhibition of the FUNDC1-mediated mitophagy in CMECs by the nuclear receptor subfamily 4 group A member 1 (NR4A1) [116] or receptor-interacting protein kinase 3 (Ripk3) [117] exhibited the disturbed mitochondrial homeostasis, upregulated the expression of EC-derived proinflammatory and adhesive factors, enhanced endothelial apoptosis, and provoked CMD in cardiac I/R injuries. In addition, enhancing autophagy through Beclin1 overexpression in CMECs exhibited suppressed NACHT, LRR, and PYD domain-containing protein 3 (NLRP3) inflammasome activation by promoting tumor necrosis factor-alpha-induced protein 3 (TNFAIP3) [118] and inhibited caspase-4 inflammasome activation [119]. Thus, it resulted in a reduced IL-1β level and an increased animal survival upon myocardial I/R injuries.…”

“…The following supporting information can be downloaded at: https: //www.mdpi.com/article/10.3390/cells11132081/s1, Table S1: Mechanistic studies implicating endothelial autophagy or mitophagy in cardiovascular diseases [93,105,106,[108][109][110][111][112][113][114][115][116][117][118][119][120][121][122][123][124][125][126][130][131][132][133][149][150][151][152][153][154][155][156][157][158][159][160][166][167][168][169][170]181,[184][185][186][187]…”

Cardiac Remodeling in Heart Failure: Role of Pyroptosis and Its Therapeutic Implications

“…NLRP3 markedly promotes pyroptosis in the progression of AMI, knockdown of NLRP3 attenuated cardiomyocyte pyroptosis and significantly decreased the infarct size, as evidenced by decreased expression levels of ASC, pro-Caspase-1, Caspase-1-p10, GSDMD, cleaved GSDMD, and IL-18 ( 45 ). Becn1-driven autophagy is a protective response in the heart during I/R, Becn1 overexpression suppressed Caspase-4 inflammasome activation and pyroptosis, alleviated microvascular damage, reduced infarct size, and mitigated cardiac inflammation and cell death ( 97 ). The recent research showed that activation of SIRT1 by inhibition of miR-29a inhibited oxidative stress, pyroptosis and protect I/R injury ( 98 , 99 ).…”

“…Becn1 overexpression increased Ischemia-reperfusion mouse survival and decreased the levels of serum LDH and CK. BECN1 attenuated F4/80+ macrophages and CD11b+ neutrophils infiltration in the heart ( 97 ). Meanwhile, myocardial fibrosis was markedly ameliorated, the collagen was decreased through suppressing GSDMD-mediated pyroptosis when PVT1 knockdown ( 96 ).…”

Pyroptosis in Myocardial Ischemia/Reperfusion Injury: Role of Endoplasmic Reticulum Stress and STING-IRF3 Pathway

“…16 Currently, gasdermins consist of six members: gasdermins A, B, C, D, E, and DFNB59. 17 Most GSDMs members have conservative N-and C-terminal structures and there are differences in the linker region. Activated caspase-1 or caspase-11/4/5 are cleaved in the linker in GSDMD to release the N-terminus with pore-forming domain (PFD) activity.…”