Molecular network, pathway, and functional analysis of time-dependent gene changes related to cathepsin G exposure in neonatal rat cardiomyocytes

Abstract

The molecular pathways activated in response to acute cathepsin G (CG) exposure, as well as the mechanisms involved in activation of signaling pathways that culminate in myocyte detachment and apoptosis remain unclear. This study aimed to determine the changes in gene expression patterns associated with time dependent CG exposure to neonatal rat cardiomyocytes (NRCMs). Microarray analysis revealed a total of 451, 572 and 1127 differentially expressed genes after CG exposure at 1, 4 and 8 h respectively. A total of 54 overlapped genes at each time point were mapped by Ingenuity Pathway Analysis (IPA). The top up-regulated genes included Hamp, SMAD6, NR4A1, FOSL2, ID3 and SLAMF7, and down-regulated genes included CYR61, GDF6, Olr640, Vom2r36, DUSP6 and MMP20. Our data suggest that there are multiple deregulated pathways associated with cardiomyocyte death after CG exposure, including JAK/Stat signaling, IL-9 signaling and Nur77 signaling. In addition, we also generated the molecular network of expressed gene and found most of the molecules were connected to ERK1/2, caspase, BCR (complex) and Cyclins. Our study reveals the ability to assess time-dependent changes in gene expression patterns in NRCMs associated with CG exposure. The global gene expression profiles may provide insight into the cellular mechanism that regulates CG dependent myocyte apoptosis. In future, the pathways important in CG response, as well as the genes found to be differentially expressed might represent the therapeutic targets for myocyte survival in heart failure.

Slit2 Protects Hearts Against Ischemia-Reperfusion Injury by Inhibiting Inflammatory Responses and Maintaining Myofilament Contractile Properties

“…It has previously been shown that cleaved Slamf7 promotes the growth of multiple myeloma (Kikuchi et al, 2020) and that inhibition of SLAMF7 blocks adhesion of multiple myeloma cells to bone marrow stromal cells (Bouchon et al, 2001;Hsi et al, 2008). Slamf7 has been found to regulate PLCγ and PI 3 K in human NK cells (Tassi and Colonna, 2005) and to be upregulated in cultured neonatal cardiomyocytes following acute cathepsin G exposure (Shukla et al, 2018). These findings suggest that Slamf7 can be present in cardiomyocytes and that it responds to various stresses.…”

Potential cardiotoxicity induced by Euodiae Fructus: In vivo and in vitro experiments and untargeted metabolomics research

“…Given the limitations of H9c2 cardiomyocytes, neonatal rat cardiomyocytes (NRCMs), considered common models for studying the morphological, biochemical, and electrophysiological characteristics of the heart ( Chlopcikova et al, 2001 ), were obtained from 2-3 day-old Sprague–Dawley (SD) rats (Beijing Si Pei Fu Biotechnology, Certificate SCXK-2019-0010) after strict sterilization by the methodology used for isolation and cultivation in previous research, with some modifications ( Sabri et al, 2003 ; Rafiq et al, 2006 ; Shukla et al, 2018 ). The apex of isolated heart tissue was digested repeatedly in the short term in a mixture of collagenase II (Biorigin Inc., China) and 0.25% trypsin (Gibco Life Technologies, China) with a magnetic stirrer at 37°C.…”