Time course of gene expression in rat experimental autoimmune myocarditis

Abstract

Genetic responses that characterize experimental autoimmune myocarditis (EAM) have not yet been determined. To investigate gene expression in the myocardium of EAM, absolute copy numbers of 44 mRNA species [calcium-handling proteins, contractile proteins, natriuretic peptides (NPs), cytokines, chemokines, growth factors, renin-angiotensin-aldosterone (RAA) system, endothelins (ETs) and extracellular matrix] in synthesized cDNA from a fixed quantity of total heart RNA were assessed using real-time reverse-transcriptase PCR at days 0, 14, 21 and 28 after immunization. alpha-Cardiac myosin showed a 26.3-fold decrease and beta-cardiac myosin a 3.75-fold increase at day 14. Atrial NP and brain NP increased 47.7- and 6.35-fold at days 21 and 14 respectively. Angiotensin II type 1 receptor, angiotensin-converting enzyme and ET1 increased 22.3-fold at day 21, 6.30-fold at day 21 and 16.8-fold at day 14 respectively. Aldosterone receptor decreased 2.15-fold at day 14, but aldosterone synthetase was detected only at days 14 and 21. Interleukin (IL)-2, IL-10, interferon-gamma and monocyte chemo-attractant protein-1 increased 9.08-fold at day 14, 398-fold at day 21, 43.1-fold at day 14 and 142-fold at day 14 respectively. Collagen type 3, collagen type 1 and fibronectin increased 34.6-, 1.74- and 44.4-fold respectively at day 21. Interestingly, osteopontin showed a 4540-fold increase and it was the highest mRNA of all at day 14. An isoform of cardiac myosin and NP are dramatically changed in EAM. RAA system and ET expressions are changed differently during the EAM time course. Cytokine, chemokine and extracellular matrix greatly increase and, in particular, large numbers of osteopontin mRNA are expressed in early EAM.

Effect of Hydrodynamics-Based Gene Delivery of Plasmid DNA Encoding Interleukin-1 Receptor Antagonist-Ig for Treatment of Rat Autoimmune Myocarditis

“…In EAM, which is a T cell-mediated disease, 27 Th1 cytokines such as IL-2 and IFN-␥ produced by CD4 ϩ T cells are thought to play a central role. 4 Therefore, inhibition of Th1 cytokine production by IL-1RA-Ig may improve EAM.…”

“…The absolute copy numbers of particular transcripts were calculated by LightCycler software using a standard curve approach. 4 cDNA from cultivated cells was then subjected to quantitative RT-PCR analysis, with the level of ␥-actin mRNA acting as an internal control.…”

“…3 Various cytokines have been found in EAM-affected hearts. 4 Interleukin-1 (IL-1), formerly known as lymphocyte activating factor, is an important inflammatory cytokine produced by monocytes, macrophages, dendritic cells, B cells, or NK cells. 5,6 There are 2 structurally distinct forms of IL-1, IL-1␣ and IL-1␤, which are both potent stimulators of target cells.…”

The effect of hydrodynamics-based delivery of an IL-13-Ig fusion gene for experimental autoimmune myocarditis in rats and its possible mechanism

“…The absolute copy numbers of all samples were calculated by LightCycler software using this standard curve [3].…”

“…EAM has been shown to be a T cell-mediated autoimmune myocarditis [2], and Th1 and Th2 cytokine balance is thought to be important for controlling progression and repair in EAM. Th1 and Th2 cytokines are produced at different stages of EAM [3]. mRNA for the Th1 cytokines IL-2 and IFN-c are expressed in the acute phase of EAM in the heart, with the Th2 cytokine IL-10 increasing later.…”