Unique transcriptional profile of sustained ligand-activated preconditioning in pre- and post-ischemic myocardium

Kevin J Ashton; Amanda Tupicoff; Grant Williams-Pritchard; Can J Kiessling; Louise E See Hoe; John P Headrick; Jason N Peart

doi:10.1371/journal.pone.0072278

Unique transcriptional profile of sustained ligand-activated preconditioning in pre- and post-ischemic myocardium

PLoS One. 2013 Aug 21;8(8):e72278. doi: 10.1371/journal.pone.0072278. eCollection 2013.

Authors

Kevin J Ashton¹, Amanda Tupicoff, Grant Williams-Pritchard, Can J Kiessling, Louise E See Hoe, John P Headrick, Jason N Peart

Affiliation

¹ Faculty of Health Sciences and Medicine, Bond University, Queensland, Australia. keashton@bond.edu.au

Abstract

Background: Opioidergic SLP (sustained ligand-activated preconditioning) induced by 3-5 days of opioid receptor (OR) agonism induces persistent protection against ischemia-reperfusion (I-R) injury in young and aged hearts, and is mechanistically distinct from conventional preconditioning responses. We thus applied unbiased gene-array interrogation to identify molecular effects of SLP in pre- and post-ischemic myocardium.

Methodology/principal findings: Male C57Bl/6 mice were implanted with 75 mg morphine or placebo pellets for 5 days. Resultant SLP did not modify cardiac function, and markedly reduced dysfunction and injury in perfused hearts subjected to 25 min ischemia/45 min reperfusion. Microarray analysis identified 14 up- and 86 down-regulated genes in normoxic hearts from SLP mice (≥1.3-fold change, FDR≤5%). Induced genes encoded sarcomeric/contractile proteins (Myh7, Mybpc3,Myom2,Des), natriuretic peptides (Nppa,Nppb) and stress-signaling elements (Csda,Ptgds). Highly repressed genes primarily encoded chemokines (Ccl2,Ccl4,Ccl7,Ccl9,Ccl13,Ccl3l3,Cxcl3), cytokines (Il1b,Il6,Tnf) and other proteins involved in inflammation/immunity (C3,Cd74,Cd83, Cd86,Hla-dbq1,Hla-drb1,Saa1,Selp,Serpina3), together with endoplasmic stress proteins (known: Dnajb1,Herpud1,Socs3; putative: Il6, Gadd45g,Rcan1) and transcriptional controllers (Egr2,Egr3, Fos,Hmox1,Nfkbid). Biological themes modified thus related to inflammation/immunity, together with cellular/cardiovascular movement and development. SLP also modified the transcriptional response to I-R (46 genes uniquely altered post-ischemia), which may influence later infarction/remodeling. This included up-regulated determinants of cellular resistance to oxidant (Mgst3,Gstm1,Gstm2) and other forms of stress (Xirp1,Ankrd1,Clu), and repression of stress-response genes (Hspa1a,Hspd1,Hsp90aa,Hsph1,Serpinh1) and Txnip.

Conclusions: Protection via SLP is associated with transcriptional repression of inflammation/immunity, up-regulation of sarcomeric elements and natriuretic peptides, and modulation of cell stress, growth and development, while conventional protective molecules are unaltered.

Publication types

Comparative Study
Research Support, Non-U.S. Gov't

MeSH terms

Animals
Gene Expression Profiling*
Ischemic Preconditioning*
Ligands
Male
Mice
Mice, Inbred BALB C
Myocardial Ischemia / genetics*
Myocardial Ischemia / physiopathology
Polymerase Chain Reaction
Transcription, Genetic*

Substances

Ligands

Grants and funding

This research was supported by grants from the National Heart Foundation of Australia and the National Health and Medical Research Council of Australia. JNP was the recipient of a Future Fellowship from the Australian Research Council. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.