Nitric oxide inhibition of transforming growth factor-beta and collagen synthesis in mesangial cells.
Culture of mesangial cells (MCs) in 5.6 vs. 30.0 mmol/l glucose for 3 weeks induced a sustained increase in protein kinase C (PKC) activity, transforming growth factor (TGF)-beta1 mRNA, bioactive TGF-beta, and collagen synthesis. Nitric oxide (NO), generated exogenously by the NO donor S-nitroso-N-acetyl, D,L-penicillamine (SNAP) or endogenously after the exposure of MC to interleukin-1beta (IL-1beta), suppressed bioactive TGF-beta in MCs cultured in 5.6 or 30.0 mmol/l glucose and suppressed or abolished increases in TGF-beta1 mRNA and collagen synthesis induced by high concentrations of glucose or phorbol 12,13-dibutyrate without altering values obtained with normal glucose concentrations. SNAP had a transient suppressive effect on PKC activity, which may explain at least in part some of the actions of SNAP. The selective inhibitor of PKC, bisindolylmaleimide (GFX), mimicked NO action. The ability of SNAP and IL-1beta to suppress TGF-beta and collagen synthesis was not mediated by cGMP, since the cGMP analog, 8-Br-PET-cGMP, did not mimic NO action and an antagonist of cGMP-dependent protein kinase, Rp-8-pCPT-cGMPs, did not prevent the inhibitory actions of SNAP. N-omega-L-arginine methyl ester (NMMA) increased TGF-beta in glomerular capillary endothelial cells (GCECs) and stimulated collagen synthesis by MC in a co-culture with GCECs. Captopril inhibited TGF-beta and collagen synthesis and increased cGMP in co-cultures of GCECs and MCs. These effects of captopril were abolished by NMMA, implying mediation by NO. Thus, endogenous NO produced by GCECs may modulate TGF-beta production by both GCECs and MCs and act to suppress matrix protein synthesis by MCs.
Craven
P A
PA
Department of Medicine, University of Pittsburgh, Pennsylvania, USA. pcraven@unixs.cis.pitt.edu
Studer
R K
RK
Felder
J
J
Phillips
S
S
DeRubertis
F R
FR
D000818:Animals; D002216:Captopril / Q000494:pharmacology; D002417:Cattle; D002478:Cells, Cultured; D018920:Coculture Techniques; D003094:Collagen / Q000096:biosynthesis* / Q000187:drug effects; D006152:Cyclic GMP / Q000494:pharmacology; D004305:Dose-Response Relationship, Drug; D004730:Endothelium, Vascular / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D004791:Enzyme Inhibitors / Q000494:pharmacology; D005920:Glomerular Mesangium / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D005947:Glucose / Q000737:chemistry / Q000494:pharmacology; D007211:Indoles / Q000494:pharmacology; D008301:Maleimides / Q000494:pharmacology; D009569:Nitric Oxide / Q000378:metabolism*; D009994:Osmolar Concentration; D010396:Penicillamine / Q000031:analogs & derivatives* / Q000378:metabolism / Q000494:pharmacology; D011392:Proline / Q000032:analysis / Q000378:metabolism; D011493:Protein Kinase C / Q000037:antagonists & inhibitors / Q000187:drug effects / Q000502:physiology*; D012333:RNA, Messenger / Q000096:biosynthesis; D026423:S-Nitroso-N-Acetylpenicillamine; D013997:Time Factors; D016212:Transforming Growth Factor beta / Q000096:biosynthesis* / Q000187:drug effects / Q000235:genetics / Q000378:metabolism; D014316:Tritium; D019323:omega-N-Methylarginine / Q000494:pharmacology
D004791:Enzyme Inhibitors; D007211:Indoles; D008301:Maleimides; D012333:RNA, Messenger; D016212:Transforming Growth Factor beta; D014316:Tritium; D019323:omega-N-Methylarginine; D009569:Nitric Oxide; D026423:S-Nitroso-N-Acetylpenicillamine; D003094:Collagen; D011392:Proline; D002216:Captopril; D011493:Protein Kinase C; D010396:Penicillamine; D006152:Cyclic GMP; D005947:Glucose; C088060:bisindolylmaleimide