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Regulation of NADPH Oxidase Mediated Oxidative Stress in Macrophages
Author(s) -
Sarna Lindsei Kathryn,
Wu Nan,
Siow Yaw L.,
Pierce Grant,
Karmin O
Publication year - 2010
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.24.1_supplement.1001.9
Subject(s) - nadph oxidase , berberine , oxidative stress , oxidase test , chemistry , intracellular , lipopolysaccharide , transfection , viability assay , superoxide , microbiology and biotechnology , biochemistry , enzyme , cell , biology , immunology , gene
Oxidative stress contributes to the pathogenesis of human diseases including atherosclerosis. The NADPH oxidase is a key source of oxidative stress in atherogenesis. Our objectives were to determine the effect of berberine, herbal medicinal compound, on NADPH oxidase‐mediated production in monocyte‐derived macrophage (THP‐1). Cells were pretreated with berberine (10–50umol/L) followed by the addition of lipopolysaccharide (LPS) (10ug/L) to induce activation of NADPH oxidase. Treatment with LPS significantly increased intracellular O 2 − over a 24 h period. Pre‐incubation with berberine resulted in a concentration‐dependent inhibition of O 2 − generation in these cells. Real‐time PCR analysis revealed an increased expression of all NADPH oxidase subunits in LPS‐treated cells. However, pretreatment with berberine (25umol/L) display a significant reduction in gp91 phox mRNA expression. Transfection with gp91 phox siRNA abolished LPS‐induced O 2 − generation in macrophages, confirming the essentiality of this subunit to oxidase activation. Cell viability tests confirmed that berberine, at concentrations sufficient for inhibiting NADPH oxidase‐mediated O 2 − generation in macrophages, did not affect cell viability. In conclusion, berberine can effectively regulate NADPH oxidase activity in stimulated macrophages by selectively inhibiting the expression of the catalytic subunit gp91 phox .