Garlic attenuates nitric oxide production in rat cardiac myocytes through inhibition of inducible nitric oxide synthase and the arginine transporter CAT-2 (cationic amino acid transporter-2).
It is now accepted that allicin, the main biologically active compound in garlic, exhibits antioxidant activity. The present study was designed to test the hypothesis that the antioxidant activity of garlic can be partially attributed to the inhibition of nitric oxide (NO) production by cytokine-induced NO synthase (iNOS). Cardiac myocytes cultured from neonatal Wistar rats were stimulated by lipopolysaccharide (LPS) and incubated for 24 h with various concentrations of allicin. This resulted in marked inhibition of nitrite production. Interestingly, a low concentration of allicin (10 microM) was significantly more potent in abrogating the effect of LPS on nitrite production than a higher concentration (40 microM). Allicin decreased steady-state iNOS mRNA levels, and this effect was maximal when a lower concentration was used (10 microM compared with 40 microM). In order to explore additional effects of allicin on NO generation that might counteract the effect on iNOS, we assessed the effects of higher allicin concentrations on arginine transport. Allicin inhibited the uptake of 1 mM extracellular arginine in a concentration-dependent manner. The expression of the two arginine transporters that are expressed in cardiac myocytes [CAT-1 (cationic amino acid transporter-1) and CAT-2] was studied using reverse transcription-PCR. A concentration of 200 microM allicin abolished the expression of CAT-2 mRNA, 100 microM significantly attenuated it, whereas 50 microM had no effect. Allicin had no effect on steady-state CAT-1 mRNA levels. Our results suggest that allicin inhibits iNOS activity through two different mechanisms: at lower concentrations it decreases iNOS mRNA levels, whereas at higher concentrations it inhibits arginine transport through down-regulation of CAT-2 mRNA.
Schwartz
Idit F
IF
Department of Nephrology, Tel Aviv Sourasky Medical Center, 6 Weizman St, Tel Aviv 64239, Israel. nmdri@netvision.net.il
Hershkovitz
Rami
R
Iaina
Adrian
A
Gnessin
Ehud
E
Wollman
Yoram
Y
Chernichowski
Tamara
T
Blum
Miriam
M
Levo
Yoram
Y
Schwartz
Doron
D
D000818:Animals; D001120:Arginine / Q000378:metabolism; D027201:Cationic Amino Acid Transporter 1 / Q000235:genetics; D027202:Cationic Amino Acid Transporter 2 / Q000037:antagonists & inhibitors* / Q000235:genetics; D002478:Cells, Cultured; D004220:Disulfides; D004305:Dose-Response Relationship, Drug; D005737:Garlic; D005786:Gene Expression Regulation / Q000187:drug effects; D008070:Lipopolysaccharides / Q000494:pharmacology; D009206:Myocardium / Q000166:cytology / Q000378:metabolism*; D009569:Nitric Oxide / Q000096:biosynthesis*; D019001:Nitric Oxide Synthase / Q000037:antagonists & inhibitors*; D052247:Nitric Oxide Synthase Type II; D012333:RNA, Messenger / Q000235:genetics; D051381:Rats; D017208:Rats, Wistar; D013441:Sulfinic Acids / Q000494:pharmacology*
D027201:Cationic Amino Acid Transporter 1; D027202:Cationic Amino Acid Transporter 2; D004220:Disulfides; D008070:Lipopolysaccharides; D012333:RNA, Messenger; D013441:Sulfinic Acids; D009569:Nitric Oxide; C006452:allicin; D001120:Arginine; D019001:Nitric Oxide Synthase; D052247:Nitric Oxide Synthase Type II; C496318:Nos2 protein, rat