Time course of gene expression in rat experimental autoimmune myocarditis

2002 ◽  
Vol 103 (6) ◽  
pp. 623-632 ◽  
Author(s):  
Haruo HANAWA ◽  
Satoru ABE ◽  
Manabu HAYASHI ◽  
Tsuyoshi YOSHIDA ◽  
Kaori YOSHIDA ◽  
...  
Keyword(s):  
Gene Expression ◽  
Extracellular Matrix ◽  
Time Course ◽  
Collagen Type ◽  
Fold Increase ◽  
Cardiac Myosin ◽  
Autoimmune Myocarditis ◽  
Experimental Autoimmune Myocarditis ◽  
Experimental Autoimmune

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. α-Cardiac myosin showed a 26.3-fold decrease and β-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-γ 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.

scholarly journals RIP1/RIP3/MLKL Mediates Myocardial Function Through Necroptosis in Experimental Autoimmune Myocarditis

2021 ◽  
Vol 8 ◽  
Author(s):  
Yujing Wu ◽  
Zhenzhong Zheng ◽  
Xiantong Cao ◽  
Qing Yang ◽  
Vikram Norton ◽  
...  
Keyword(s):  
Cell Death ◽  
Myocardial Function ◽  
Viral Myocarditis ◽  
Cardiac Myosin ◽  
Autoimmune Myocarditis ◽  
Potential Therapeutic Target ◽  
Inflammatory Infiltration ◽  
Experimental Autoimmune Myocarditis ◽  
Experimental Autoimmune

Cardiomyopathy often leads to dilated cardiomyopathy (DCM) when caused by viral myocarditis. Apoptosis is long considered as the principal process of cell death in cardiomyocytes, but programmed necrosis or necroptosis is recently believed to play an important role in cardiomyocyte cell death. We investigated the role of necroptosis and its interdependency with other processes of cell death, autophagy, and apoptosis in a rat system of experimental autoimmune myocarditis (EAM). We successfully created a rat model system of EAM by injecting porcine cardiac myosin (PCM) and showed that in EAM, all three forms of cell death increase considerably, resulting in the deterioration of cardiac conditions with an increase in inflammatory infiltration in cardiomyocytes. To explore whether necroptosis occurs in EAM rats independent of autophagy, we treated EAM rats with a RIP1/RIP3/MLKL kinase-mediated necroptosis inhibitor, Necrostatin-1 (Nec-1). In Nec-1 treated rats, cell death proceeds through apoptosis but has no significant effect on autophagy. In contrast, autophagy inhibitor 3-Methyl Adenine (3-MA) increases necroptosis, implying that blockage of autophagy must be compensated through necroptosis. Caspase 8 inhibitor zVAD-fmk blocks apoptosis but increases both necroptosis and autophagy. However, all necroptosis, apoptosis, and autophagy inhibitors independently reduce inflammatory infiltration in cardiomyocytes and improve cardiac conditions. Since apoptosis or autophagy is involved in many important cellular aspects, instead of suppressing these two major cell death processes, Nec1 can be developed as a potential therapeutic target for inflammatory myocarditis.

scholarly journals Gene expression and extracellular matrix ultrastructure of a mineralizing chondrocyte cell culture system.

1991 ◽  
Vol 112 (3) ◽  
pp. 501-513 ◽  
Author(s):  
L C Gerstenfeld ◽  
W J Landis
Keyword(s):  
Extracellular Matrix ◽  
Culture System ◽  
Time Course ◽  
Collagen Synthesis ◽  
Collagen Type ◽  
Fold Increase ◽  
Proteoglycan Synthesis ◽  
Type Ii ◽  
Collagen Types ◽  
Matrix Mineralization

Conditions were defined for promoting cell growth, hypertrophy, and extracellular matrix mineralization of a culture system derived from embryonic chick vertebral chondrocytes. Ascorbic acid supplementation by itself led to the hypertrophic phenotype as assessed by respective 10- and 15-fold increases in alkaline phosphatase enzyme activity and type X synthesis. Maximal extracellular matrix mineralization was obtained, however, when cultures were grown in a nutrient-enriched medium supplemented with both ascorbic acid and 20 mM beta-glycerophosphate. Temporal studies over a 3-wk period showed a 3-4-fold increase in DNA accompanied by a nearly constant DNA to protein ratio. In this period, total collagen increased from 3 to 20% of the cell layer protein; total calcium and phosphorus contents increased 15-20-fold. Proteoglycan synthesis was maximal until day 12 but thereafter showed a fourfold decrease. In contrast, total collagen synthesis showed a greater than 10-fold increase until day 18, a result suggesting that collagen synthesis was replacing proteoglycan synthesis during cellular hypertrophy. Separate analysis of individual collagen types demonstrated a low level of type I collagen synthesis throughout the 21-d time course. Collagen types II and X synthesis increased during the first 2 wk of culture; thereafter, collagen type II synthesis decreased while collagen type X synthesis continued to rise. Type IX synthesis remained at undetectable levels throughout the time course. The levels of collagen types I, II, IX, and X mRNA and the large proteoglycan core protein mRNA paralleled their levels of synthesis, data indicating pretranslational control of synthesis. Ultrastructural examination revealed cellular and extracellular morphology similar to that for a developing hypertrophic phenotype in vivo. Chondrocytes in lacunae were surrounded by a well-formed extracellular matrix of randomly distributed collagen type II fibrils (approximately 20-nm diam) and extensive proteoglycan. Numerous vesicular structures could be detected. Cultures mineralized reproducibly and crystals were located in extracellular matrices, principally associated with collagen fibrils. There was no clear evidence of mineral association with extracellular vesicles. The mineral was composed of calcium and phosphorus on electron probe microanalysis and was identified as a very poorly crystalline hydroxyapatite on electron diffraction. In summary, these data suggest that this culture system consists of chondrocytes which undergo differentiation in vitro as assessed by their elevated levels of alkaline phosphatase and type X collagen and their ultrastructural appearance.(ABSTRACT TRUNCATED AT 400 WORDS)

scholarly journals Treatment with N-acetyl-seryl-aspartyl-lysyl-proline prevents experimental autoimmune myocarditis in rats

2012 ◽  
Vol 303 (9) ◽  
pp. H1114-H1127 ◽  
Author(s):  
Pablo Nakagawa ◽  
Yunhe Liu ◽  
Tang-Dong Liao ◽  
Xiaojuan Chen ◽  
Germán E. González ◽  
...  
Keyword(s):  
T Cell ◽  
Immune Responses ◽  
Adhesion Molecules ◽  
Cardiac Dysfunction ◽  
Myocardial Injury ◽  
Cardiac Myosin ◽  
Autoimmune Myocarditis ◽  
Cell Responses ◽  
Experimental Autoimmune Myocarditis ◽  
Experimental Autoimmune

Myocarditis is commonly associated with cardiotropic infections and has been linked to development of autoimmunity. N-acetyl-seryl-aspartyl-lysyl-proline (Ac-SDKP) is a naturally occurring tetrapeptide that prevents inflammation and fibrosis in hypertension and other cardiovascular diseases; however, its effect on autoimmune-mediated cardiac diseases remains unknown. We studied the effects of Ac-SDKP in experimental autoimmune myocarditis (EAM), a model of T cell-mediated autoimmune disease. This study was conducted to test the hypothesis that Ac-SDKP prevents autoimmune myocardial injury by modulating the immune responses. Lewis rats were immunized with porcine cardiac myosin and treated with Ac-SDKP or vehicle. In EAM, Ac-SDKP prevented both systolic and diastolic cardiac dysfunction, remodeling as shown by hypertrophy and fibrosis, and cell-mediated immune responses without affecting myosin-specific autoantibodies or antigen-specific T cell responses. In addition, Ac-SDKP reduced cardiac infiltration by macrophages, dendritic cells, and T cells, pro-inflammatory cytokines [interleukin (IL)-1α, tumor necrosis factor-α, IL-2, IL-17] and chemokines (cytokine-induced neutrophil chemoattractant-1, interferon-γ-induced protein 10), cell adhesion molecules (intercellular adhesion molecule-1, L-selectin), and matrix metalloproteinases (MMP). Ac-SDKP prevents autoimmune cardiac dysfunction and remodeling without reducing the production of autoantibodies or T cell responses to cardiac myosin. The protective effects of Ac-SDKP in autoimmune myocardial injury are most likely mediated by inhibition of 1) innate and adaptive immune cell infiltration and 2) expression of proinflammatory mediators such as cytokines, chemokines, adhesion molecules, and MMPs.

Myocarditis triggers inflammatory response in cardiac fibroblasts and profibrotic activation of myeloid and endothelial cells in mouse model of experimental autoimmune myocarditis

2020 ◽  
Vol 41 (Supplement_2) ◽  
Author(s):  
K Tkacz ◽  
E Dzialo ◽  
K Weglarczyk ◽  
M Czepiel ◽  
M Siedlar ◽  
...  
Keyword(s):  
Endothelial Cells ◽  
Collagen Type ◽  
Cardiac Fibroblasts ◽  
Myeloid Cells ◽  
Collagen Type I ◽  
Funding Source ◽  
Type I ◽  
Autoimmune Myocarditis ◽  
Experimental Autoimmune Myocarditis ◽  
Experimental Autoimmune

Abstract Background/Introduction Myocarditis, a heart-specific inflammation, is a common cause of pathological tissue remodeling and cardiac fibrosis resulting in stiffening of ventricles, functional impairment and heart failure. Immunization of susceptible mice with alpha myosin heavy chain (αMyHC) and complete Freund's adjuvant (CFA) induces CD4+ T cell-mediated experimental autoimmune myocarditis (EAM). In EAM model, resolution of acute cardiac inflammation is followed by a progressive dilated cardiomyopathy and systolic dysfunction. Purpose The aim of our study was to identify the role of resident cardiac fibroblasts, cardiac endothelial as well as inflammatory myeloid cells during the course of EAM. Methods EAM was induced by immunization with αMyHC/CFA in reporter BALB/c mice expressing EGFP under collagen type I promoter (Coll-EGFP) and RFP under a control of α-smooth muscle actin (αSMA) promoter (αSMA-RFP). Using flow cytometry analysis, cardiac cells were phenotyped and quantified at inflammatory (d19–21) and fibrotic (d40) stage of EAM. Sorted EGFP-positive cardiac fibroblasts obtained from healthy and myocarditis-positive mice (day 21 of EAM) were comparatively analyzed for the whole genome transcriptomics using the Next Generation Sequencing with read length 2x150bp and 20–30 million reads per sample. Results A massive infiltration of inflammatory CD45+CD11b+ myeloid cells (mainly CD11b+CD36+ macrophages, CD11b+CD36–Ly6GhiLy6chi neutrophils, CD11b+CD36–Ly6G–Ly6c– monocytes, CD11b+CD36–Ly6G–Ly6chi inflammatory monocytes) was observed at day 21 of EAM. Myeloid cells as well as endothelial cells showed increased production of type I collagen at day 21, which was further reduced at day 40 of EAM. At day 21, collagen-producing endothelial cells showed particularly elevated levels of adhesion molecules ICAM and VCAM. On the other hand, the total number of EGFP-positive cardiac fibroblasts remained unchanged during the course of EAM, as well as the percentage of cardiac fibroblasts positive for αSMA (myofibroblasts). Gene ontology analysis of transcripts differentially regulated in cardiac fibroblasts during acute myocarditis pointed mainly to activation of immune processes, response to stress, cytoskeletal and extracellular matrix organization. Specifically, in EAM at day 21 cardiac fibroblasts increased transcription of chemokines (Ccl6, Ccl9, Cxcl2, Cxcl3, Cxcl5, Cxcl9, Cxcl13), collagens (Col6a4, Col6a5, Col9a1, Col9a3, Col11a2, Col12a1, Col24a1, Col28a1), and genes involved in ECM biology (Bmp7, Kng2, Lgals3, Cthrc1, Cela1, Spn). Conclusions In EAM model, inflammatory myeloid and cardiac endothelial cells seem to contribute to excessive collagen type I production, whereas cardiac fibroblasts actively participate in inflammatory and profibrotic responses. Funding Acknowledgement Type of funding source: Public grant(s) – National budget only. Main funding source(s): The National Science Centre (Poland)

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