Anti-inflammatory effect of heme oxygenase 1: Glycosylation and nitric oxide inhibition in macrophages
Article first published online: 12 AUG 2004
Copyright © 2004 Wiley-Liss, Inc.
Journal of Cellular Physiology
Volume 202, Issue 2, pages 579–590, February 2005
How to Cite
Lin, H.-Y., Shen, S.-C. and Chen, Y.-C. (2005), Anti-inflammatory effect of heme oxygenase 1: Glycosylation and nitric oxide inhibition in macrophages. J. Cell. Physiol., 202: 579–590. doi: 10.1002/jcp.20160
- Issue published online: 23 NOV 2004
- Article first published online: 12 AUG 2004
- Manuscript Accepted: 19 MAY 2004
- Manuscript Received: 4 MAY 2004
- National Science Council (NSC). Grant Numbers: 91-2320-B-038-040, 92-2320-B-038-021
Flavonoids including the aglycones, hesperetin (HT; 5,7,3′-trihydroxy-4′-methoxy-flavanone), and naringenin (NE; 5,7,4′-trihydroxy flavanone) and glycones, hesperidin (HD; 5,7,3′-trihydroxy-4′-methoxy-flavanone 7-rhamnoglucoside) and naringin (NI; 5,7,4′-trihydroxy flavanone 7-rhamno glucoside), were used to examine the importance of rutinose at C7 on the inhibitory effects of flavonoids on lipopolysaccharide (LPS)-induced nitric oxide production in macrophages. Both HT and NE, but not their respective glycosides HD and NI, induced heme oxygenase 1 (HO-1) protein expression in the presence or absence of LPS and showed time and dose-dependent inhibition of LPS-induced nitric oxide (NO) production and inducible nitric oxide synthase (iNOS) expression in RAW264.7, J774A.1, and thioglycolate-elicited peritoneal macrophages. Additive inhibitory effect of an HO-1 inducer hemin and NE or NI on LPS-induced NO production and iNOS expression was identified, and HO enzyme inhibitor tin protoporphyrin (SnPP) attenuated the inhibitory effects of HT, NE, and hemin on LPS-induced NO production. Both NE and HT showed no effect on iNOS mRNA and protein stability in RAW264.7 cells. Removal of rutinose at C7 of HD and NI by enzymatic digestion using hesperidinase (HDase) and naringinase (NIase) produce inhibitory activity on LPS-induced NO production, according to the production of the aglycones, HT and NE, by high-performance liquid chromatography (HPLC) analysis. Furthermore, the amount of NO produced by LPS or lipoteichoic acid (LTA) was significantly reduced in HO-1-overexpressing cells (HO-1/RAW264.7) compared to that in parental cells (RAW264.7). Results of the present study provide scientific evidence to suggest that rutinose at C7 is a negative moiety in flavonoid inhibition of LPS-induced NO production, and that HO-1 is involved in the inhibitory mechanism of flavonoids on LPS-induced iNOS and NO production. © 2004 Wiley-Liss, Inc.