GSK-3β-mediated activation of NLRP3 inflammasome leads to pyroptosis and apoptosis of rat cardiomyocytes and fibroblasts

• Published in: European journal of pharmacology Vol. 920; p. 174830
• Main Authors: Wang, Shu-Hui, Cui, Liu-Gen, Su, Xue-Ling, Komal, Sumra, Ni, Rui-Cong, Zang, Ming-Xi, Zhang, Li-Rong, Han, Sheng-Na
• Format: Journal Article
• Language: English
• Published: Netherlands Elsevier B.V 05-04-2022
• Subjects: Animals; Apoptosis; Fibroblasts - metabolism; Glycogen Synthase Kinase 3 beta - metabolism; GSK-3β; Inflammasomes - metabolism; Interleukin-1beta - metabolism; Myocardial infarction; Myocytes, Cardiac; NLR Family, Pyrin Domain-Containing 3 Protein - metabolism; NLRP3 inflammasome; Pyroptosis; Rats; Rats, Sprague-Dawley; Myocardial infarction; GSK-3β; Pyroptosis; NLRP3 inflammasome; Apoptosis
• ISSN: 0014-2999; 1879-0712
• DOI: 10.1016/j.ejphar.2022.174830

We previously demonstrated that GSK-3β mediates NLRP3 inflammasome activation and IL-1β production in cardiac fibroblasts (CFs) after myocardial infarction (MI). In this study, we show how GSK-3β-mediated activation of the NLRP3 inflammasome/caspase-1/IL-1β pathway leads to apoptosis and pyroptosis of cardiomyocytes (CMs) and CFs. Administration of lipopolysaccharide (LPS)/ATP to primary newborn rat cardiac fibroblasts (RCFs) led to increase in proteins of NLRP3, apoptosis-associated speck-like protein containing a caspase recruitment domain (ASC), caspase-1, IL-1β, and IL-18. Additionally, the expression of caspase-3 and N-terminal fragments of gasdermin D (N-GSDMD) and the Bax/Bcl-2 ratio increased. Administration of the GSK-3β inhibitor SB216763 reduced the levels of apoptosis- and pyroptosis-related proteins regulated by NLRP3 inflammasome activation in RCFs. Next, we transferred the culture supernatant of LPS/ATP-treated RCFs to in vitro primary newborn rat cardiomyocytes (RCMs). The results showed that SB216763 attenuate the upregulation of the ratios of Bax/Bcl-2 and the expression of caspase-3 and N-GSDMD in RCMs. Direct stimulation of RCMs and H9c2 cells with recombinant rat IL-1β increased the p-GSK-3β/GSK-3β and Bax/Bcl-2 ratios and the expression of caspase-3 and N-GSDMD, while both SB216763 and TLR1 (an IL-1β receptor inhibitor) markedly reduced these effects, as assessed using propidium iodide positive staining and the lactate dehydrogenase release assay. The caspase-11 inhibitor wedelolactone decreased the expression level of N-GSDMD but did not alter the p-GSK-3β/GSK-3β ratio. Lastly, we established a Sprague–Dawley rat MI model to confirm that SB216763 diminished the increase in caspase-3 and N-GSDMD expression and the Bax/Bcl-2 ratio in the ischemic area. These data demonstrate that GSK-3β regulates apoptosis and pyroptosis of RCMs and RCFs due to NLRP3 inflammasome activation in RCFs.