Differentially expressed genes in L6 rat skeletal muscle myoblasts after incubation with inflammatory cytokines.
OBJECTIVE
The mechanism underlying exercise intolerance in chronic heart failure is still unclear. An increased concentration of inflammatory cytokines could be detected in the serum of patients with chronic heart failure (CHF) exhibiting a correlation with the severity of the disease. The variety of molecular alterations triggered by these cytokines in the skeletal muscle is almost unknown. The study was designed to analyze the differential gene expression in skeletal muscle myoblasts after stimulation with inflammatory cytokines.


METHODS
L6 rat skeletal muscle myoblasts were incubated for 24 h with a combination of IL-1beta/IFN-gamma and the differential gene expression profile was determined by a PCR-based subtractive hybridization method.


RESULTS
Out of 173 picked clones 141 different sequences could be identified. By comparison with Genebank, the identity of 73 genes (51.7%) could be confirmed, whereas the rest did not show a homology to any known gene. Some of the identified genes are known to be altered in patients with CHF.


CONCLUSIONS
In summary, the results of this study provide information about changes in gene expression after exposure of skeletal muscle cells to inflammatory cytokines. This information may yield a new gene pool, worthwhile to be analyzed in skeletal muscle of patients with chronic heart failure.
Adams
V
V
Department of Cardiology, Heart Center, University of Leipzig, Leipzig, Germany. adav@server3.medizin.uni-leipzig.de
Lenk
K
K
Schubert
A
A
Gielen
S
S
Schuler
G
G
Hambrecht
R
R
D000818:Animals; D017209:Apoptosis; D015152:Blotting, Northern; D015153:Blotting, Western; D002478:Cells, Cultured; D003001:Cloning, Molecular; D016207:Cytokines / Q000378:metabolism*; D018076:DNA, Complementary / Q000378:metabolism; D004268:DNA-Binding Proteins / Q000096:biosynthesis; D016208:Databases, Factual; D004587:Electrophoresis, Agar Gel; D005109:Extracellular Matrix / Q000378:metabolism; D006333:Heart Failure / Q000235:genetics / Q000378:metabolism; D007371:Interferon-gamma / Q000494:pharmacology; D007375:Interleukin-1 / Q000494:pharmacology; D020928:Mitogen-Activated Protein Kinases / Q000096:biosynthesis; D018482:Muscle, Skeletal / Q000378:metabolism; D009132:Muscles / Q000166:cytology*; D019001:Nitric Oxide Synthase / Q000096:biosynthesis; D052247:Nitric Oxide Synthase Type II; D009687:Nuclear Proteins; D009693:Nucleic Acid Hybridization; D010957:Plasmids / Q000378:metabolism; D012333:RNA, Messenger / Q000378:metabolism; D051381:Rats; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013997:Time Factors; D014157:Transcription Factors; D044767:Ubiquitin-Protein Ligases; D048051:p38 Mitogen-Activated Protein Kinases
D016207:Cytokines; D018076:DNA, Complementary; D004268:DNA-Binding Proteins; D007375:Interleukin-1; D009687:Nuclear Proteins; D012333:RNA, Messenger; C114070:RNF4 protein, human; D014157:Transcription Factors; D007371:Interferon-gamma; D019001:Nitric Oxide Synthase; D052247:Nitric Oxide Synthase Type II; C496318:Nos2 protein, rat; C486411:Rnf4 protein, rat; D044767:Ubiquitin-Protein Ligases; D020928:Mitogen-Activated Protein Kinases; D048051:p38 Mitogen-Activated Protein Kinases