Gentamicin induces Jun-AP1 expression and JNK activation in renal glomeruli and cultured mesangial cells.
Reactive oxygen species (ROS) mediate MC contraction, proliferation and apoptosis induced by gentamicin (G) in vitro and in vivo. Sustained increases in cytosolic free calcium, increased iNOS expression and elevated nitric oxide (NO) production are associated with MC apoptosis in vitro. As NO strongly activated c-Jun N-terminal kinase (JNK) and increased AP1 expression, and these two factors are involved in MC proliferation in vitro, we have measured Jun-AP1 expression in rat glomeruli from G-treated rats, and the effect of G on Jun-AP1 expression and JNK activity in cultured MC. Moreover, we studied the expression of inducible (iNOS) and constitutive (cNOS) NO synthases in rat glomeruli. Glomeruli were obtained from rats treated with G (100 mg/kg body weight/day) along 6 days, and MC primary cultures were evaluated after 24, 48 and 72 h incubation with 10(-5) M G. G induced an increase in the expression of iNOS, cNOS and Jun-AP1 in rat glomeruli and in MC cultures. Moreover, G activated JNK; JNK activation was reduced by co-incubation with the calcium channel blocker verapamil and with the ROS scavengers superoxide dismutase and catalase. These results strongly suggest a role for reactive oxygen/nitrogen species produced by increased NOS activity in G-induced MC activation. These reactive oxygen molecules and increased intracellular free calcium may mediate the increase in Jun-AP1 expression and JNK activation induced by G treatment in MC.
Martínez-Salgado
Carlos
C
Unidad de Investigación, Hospital Universitario de Salamanca, Paseo de San Vicente 58-132, 37007 Salamanca, Spain. carlosms@usal.es
Rodríguez-Barbero
Alicia
A
Eleno
Nélida
N
López-Novoa
José M
JM
D000818:Animals; D015153:Blotting, Western; D002374:Catalase / Q000494:pharmacology; D002478:Cells, Cultured; D004789:Enzyme Activation / Q000187:drug effects; D005786:Gene Expression Regulation / Q000187:drug effects*; D005839:Gentamicins / Q000494:pharmacology*; D047468:Immunoprecipitation; D048031:JNK Mitogen-Activated Protein Kinases / Q000378:metabolism*; D007678:Kidney Glomerulus / Q000166:cytology / Q000378:metabolism*; D050527:Mesangial Cells / Q000378:metabolism*; D019001:Nitric Oxide Synthase / Q000378:metabolism; D051381:Rats; D017208:Rats, Wistar; D013482:Superoxide Dismutase / Q000494:pharmacology; D014157:Transcription Factors / Q000378:metabolism*; D014700:Verapamil / Q000494:pharmacology
D005839:Gentamicins; D014157:Transcription Factors; D014700:Verapamil; D002374:Catalase; D019001:Nitric Oxide Synthase; D013482:Superoxide Dismutase; D048031:JNK Mitogen-Activated Protein Kinases