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

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

  • Eur J Pharmacol. 2022 Apr 5;920:174830. doi: 10.1016/j.ejphar.2022.174830.
Shu-Hui Wang 1 Liu-Gen Cui 2 Xue-Ling Su 2 Sumra Komal 2 Rui-Cong Ni 2 Ming-Xi Zang 3 Li-Rong Zhang 4 Sheng-Na Han 5
Affiliations

Affiliations

  • 1 Department of Pharmacology, School of Basic Medical Sciences, Zhengzhou University, Zhengzhou, 450001, China; Department of Ultrasound, Affiliated Tumor Hospital of ZhengZhou University, Zhengzhou, 450008, China.
  • 2 Department of Pharmacology, School of Basic Medical Sciences, Zhengzhou University, Zhengzhou, 450001, China.
  • 3 Department of Biochemistry & Molecular Biology, School of Basic Medical Sciences, Zhengzhou University, Zhengzhou, 450001, China.
  • 4 Department of Pharmacology, School of Basic Medical Sciences, Zhengzhou University, Zhengzhou, 450001, China. Electronic address: lrzhang@zzu.edu.cn.
  • 5 Department of Pharmacology, School of Basic Medical Sciences, Zhengzhou University, Zhengzhou, 450001, China. Electronic address: hsn@zzu.edu.cn.
Abstract

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.

Keywords

Apoptosis; GSK-3β; Myocardial infarction; NLRP3 inflammasome; Pyroptosis.

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