Cytokine gene expression in Adriamycin nephropathy: effects of antioxidant nuclear factor kappaB inhibitors in established disease.
OBJECTIVE
Inhibition of nuclear factor kappaB with the antioxidant pyrrolidine dithiocarbamate (PDTC) reduced tubulointerstitial injury in Adriamycin nephropathy (AN), whereas N-acetylcysteine (NAC) was ineffective. Here we hypothesize that PDTC reduces the renal cortical expression of nuclear factor kappaB dependent cytokines in AN.


METHODS
Male Wistar rats received a single intravenous injection of doxorubicin hydrochloride (7.5 mg/kg). NAC (150 mg/kg twice daily i.p.), PDTC (50 mg/kg twice daily i.p.), or vehicle were commenced on day 14 and continued until day 30.


RESULTS
On day 30, mRNAs of selected cytokines were increased in AN (TNF-alpha 3.4-fold, MCP-1 5.1-fold, IL-10 2.7-fold, TGF-beta1 3.5-fold, all p < 0.05) as determined by RT-PCR. PDTC reduced IL-10 and TGF-beta1 mRNAs (p < 0.05), whereas the upregulation of MCP-1 and TNF-alpha mRNAs was not affected. In contrast, NAC increased TNF-alpha and IL-10 mRNAs (p < 0.05). Nuclear protein levels of activator protein-1 were increased in AN (4.4-fold, p < 0.01) and not significantly altered by PDTC (3.0-fold, p = 0.13) or NAC (5. 2-fold, p = 0.18).


CONCLUSIONS
The protective effects of PDTC in AN are not associated with a local reduction in TNF-alpha and MCP-1 gene expression. The latter may be due to continued transactivation by activator protein-1. These data also suggest that IL-10 and TGF-beta1 mRNA expressions are PDTC dependent and have a role in mediating tubulointerstitial injury.
Rangan
G K
GK
Department of Renal Medicine, University of Sydney at Westmead Hospital, Sydney, Australia. gkr@u.washington.edu
Wang
Y
Y
Tay
Y C
YC
Harris
D C
DC
D000111:Acetylcysteine / Q000494:pharmacology; D000818:Animals; D000903:Antibiotics, Antineoplastic / Q000633:toxicity*; D000975:Antioxidants / Q000494:pharmacology*; D016207:Cytokines / Q000096:biosynthesis* / Q000235:genetics; D004317:Doxorubicin / Q000633:toxicity*; D016166:Free Radical Scavengers / Q000494:pharmacology; D007674:Kidney Diseases / Q000139:chemically induced / Q000378:metabolism*; D008297:Male; D016328:NF-kappa B / Q000037:antagonists & inhibitors*; D009687:Nuclear Proteins / Q000096:biosynthesis / Q000302:isolation & purification; D011759:Pyrrolidines / Q000494:pharmacology; D012333:RNA, Messenger / Q000096:biosynthesis / Q000235:genetics; D051381:Rats; D017208:Rats, Wistar; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013859:Thiocarbamates / Q000494:pharmacology
D000903:Antibiotics, Antineoplastic; D000975:Antioxidants; D016207:Cytokines; D016166:Free Radical Scavengers; D016328:NF-kappa B; D009687:Nuclear Proteins; D011759:Pyrrolidines; D012333:RNA, Messenger; D013859:Thiocarbamates; C020972:pyrrolidine dithiocarbamic acid; D004317:Doxorubicin; D000111:Acetylcysteine