Nitric oxide modulates expression of extracellular matrix genes linked to fibrosis in kidney mesangial cells.
Mesangial cells are thought to be important mediators of glomerular inflammation and fibrosis. Studies have established a direct role for nitric oxide (NO) in the regulation of gene expression in mesangial cells. Representational difference analysis was used to investigate changes in gene expression elicited by the treatment of S-nitroso-L-glutathione in rat mesangial cells. Seven upregulated and 11 downregulated genes were identified. Four out of 11 downregulated genes (connective tissue growth factor, thrombospondin-1, collagen type I alpha1 and collagen type I alpha2) are known to be linked to inflammation and fibrosis. Results were verified across species in mesangial cells treated with a series of NO donors using Northern blot analysis, quantitative real-time PCR and protein analysis methods. Induction of endogenous NO production by cytokine stimulation also triggered regulation of the genes. One example gene, connective tissue growth factor, was studied at the promoter level. Promoter-reporter gene studies in mesangial cells demonstrated that NO acts at the transcriptional level to suppress gene expression. Our results reveal a complex role of NO in regulating gene expression in mesangial cells and suggest an antifibrotic potential for NO.
Wani
Javid
J
1Medical Policlinic, Ludwig Maximilians University, Schillerstrasse 42, Munich, Germany.
Carl
Marina
M
Henger
Anna
A
Nelson
Peter J
PJ
Rupprecht
Harald
H
D000818:Animals; D058578:Biglycan; D001708:Biopterins / Q000031:analogs & derivatives / Q000494:pharmacology; D002478:Cells, Cultured; D003094:Collagen / Q000235:genetics; D055513:Connective Tissue Growth Factor; D015536:Down-Regulation; D004789:Enzyme Activation / Q000187:drug effects; D016326:Extracellular Matrix Proteins / Q000235:genetics* / Q000378:metabolism*; D005355:Fibrosis / Q000235:genetics / Q000378:metabolism / Q000473:pathology; D017874:Immediate-Early Proteins / Q000378:metabolism; D036341:Intercellular Signaling Peptides and Proteins / Q000378:metabolism; D007371:Interferon-gamma / Q000494:pharmacology; D050527:Mesangial Cells / Q000187:drug effects / Q000378:metabolism* / Q000473:pathology*; D009569:Nitric Oxide / Q000378:metabolism*; D052247:Nitric Oxide Synthase Type II / Q000037:antagonists & inhibitors / Q000378:metabolism; D011401:Promoter Regions, Genetic / Q000235:genetics; D011509:Proteoglycans / Q000235:genetics; D020871:RNA Stability; D012333:RNA, Messenger / Q000235:genetics; D051381:Rats; D019700:Thrombospondin 1 / Q000378:metabolism
C546886:Bgn protein, rat; D058578:Biglycan; C526316:CCN2 protein, rat; D016326:Extracellular Matrix Proteins; D017874:Immediate-Early Proteins; D036341:Intercellular Signaling Peptides and Proteins; D011509:Proteoglycans; D012333:RNA, Messenger; D019700:Thrombospondin 1; D055513:Connective Tissue Growth Factor; D001708:Biopterins; D009569:Nitric Oxide; D007371:Interferon-gamma; D003094:Collagen; D052247:Nitric Oxide Synthase Type II; C003402:sapropterin