LncRNA PVT1 Knockdown Ameliorates Myocardial Ischemia Reperfusion Damage via Suppressing Gasdermin D-Mediated Pyroptosis in Cardiomyocytes

Abstract

Objective: Myocardial ischemia reperfusion (I/R) damage is a life-threatening vascular emergency after myocardial infarction. Here, we observed the cardioprotective effect of long non-coding RNA (lncRNA) PVT1 knockdown against myocardial I/R damage.Methods: This study constructed a myocardial I/R-induced mouse model and a hypoxia/reoxygenation (H/R)-treated H9C2 cells. PVT1 expression was examined via RT-qPCR. After silencing PVT1 via shRNA against PVT1, H&E, and Masson staining was performed to observe myocardial I/R damage. Indicators of myocardial injury including cTnI, LDH, BNP, and CK-MB were examined by ELISA. Inflammatory factors (TNF-α, IL-1β, and IL-6), Gasdermin D (GSDMD), and Caspase1 were detected via RT-qPCR, western blot, immunohistochemistry, or immunofluorescence. Furthermore, CCK-8 and flow cytometry were presented for detecting cell viability and apoptosis.Results: LncRNA PVT1 was markedly up-regulated in myocardial I/R tissue specimens as well as H/R-induced H9C2 cells. Silencing PVT1 significantly lowered serum levels of cTnI, LDH, BNP, and CK-MB in myocardial I/R mice. H&E and Masson staining showed that silencing PVT1 alleviated myocardial I/R injury. PVT1 knockdown significantly lowered the production and release of inflammatory factors as well as inhibited the expression of GSDMD-N and Caspase1 in myocardial I/R tissue specimens as well as H/R-induced H9C2 cells. Moreover, silencing PVT1 facilitated cell viability and induced apoptosis of H/R-treated H9C2 cells.Conclusion: Our findings demonstrated that silencing PVT1 could alleviate myocardial I/R damage through suppressing GSDMD-mediated pyroptosis in vivo and in vitro. Thus, PVT1 knockdown may offer an alternative therapeutic strategy against myocardial I/R damage.

Identification of the metabolic remodeling profile in the early-stage of myocardial ischemia and the contributory role of mitochondrion

“…The blood was centrifuged at 4000 rpm for 20 min at room temperature, and serum was remained for assay. ELISA kits of D-LDH (Ruixinbio, RX203251M, Quanzhou, China) and CK-MB (Ruixinbio, RX201007M, Quanzhou, China) were used for detecting the enzymes as previously described [ 34 , 35 ]. The absorbance was measured at 450 nm.…”

Gasdermin D-mediated pyroptosis in myocardial ischemia and reperfusion injury: Cumulative evidence for future cardioprotective strategies

“…Pyroptosis is also shown to have a role in the hypoxia/reoxygenation (H/R) model, which involves upregulation of the pro-inflammatory signaling pathway ( Table 3 12 , 19 , 21 , 45 , 46 , 47 , 48 , 49 , 50 , 51 ). H/R injury activated thioredoxin-interacting protein (TXNIP), which triggered the TLR4/myeloid differentiation factor 88 (MYD88)/NF- κ B signaling pathway, and caused NLRP3 inflammasome/caspase-1-mediated pyroptosis 45 , 46 .…”

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

“…Serum GSDMD levels were also significantly higher after percutaneous coronary intervention in patients with ST-segment–elevation myocardial infarction than in age-matched stable coronary artery disease patients ( 89 ). Moreover, trimetazidine and silencing PVT1 could alleviate myocardial I/R damage through suppressing GSDMD-mediated pyroptosis in vivo and in vitro, involved TLR4/MyD88/NF-κB/NLRP3 inflammasome pathway, improved cardiac fibrosis, inflammatory cytokines, and cardiac function ( 94 , 96 ). 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 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 ). miR-135b can alleviate the fibrosis of cardiac fibroblasts, as well as cardiac fibroblast pyroptosis, which can be inhibited via miR-135b that is directly bound to Caspase-1 ( 71 ).…”