Expression of functionally phagocyte-type NAD(P)H oxidase in pericytes: effect of angiotensin II and high glucose.
BACKGROUND
A growing body of evidence demonstrates the involvement of the oxidative stress in the development of vascular complications associated with diabetes, such as hypertension, retinopathy, nephropathy, neuropathy and atherosclerosis. However, the molecular mechanisms accountable for the increased production of reactive oxygen species (ROS) remain uncertain. Among others, the NAD(P)H oxidase is one of the most important sources of superoxide anion (O2-) that induce dysfunction of vascular cells. Pericytes (PCs) have an essential role in the capillary dysfunction in retinopathy and other vascular complications in diabetes. We questioned whether PCs express a functional phagocyte-type NAD(P)H oxidase, and examined the role of angiotensin II and high glucose on the activity of the oxidase complex and expression of the essential subunit p22(phox).


RESULTS
The mRNA expression of p22(phox), p47(phox), p67(phox) and NOX 1 subunits, and the lack of gp91(phox) component, were detected in PCs by reverse transcriptase PCR. Western-blotting analysis demonstrated the protein expression of p22(phox), p47(phox) and p67(phox) subunits. As compared with the normal condition, stimulation of PCs with angiotensin II or high glucose induced: (i) an increase in ROS production and NAD(P)H oxidase activity, and (ii) an up-regulation of p22(phox) mRNA and protein expression.


CONCLUSIONS
Taken together, the present study provides the first evidence that PCs express a functional phagocyte-type NAD(P)H oxidase, which is up-regulated by both angiotensin II and high glucose. Given the importance of ROS in vascular physiology and pathology, the NAD(P)H oxidase complex could be an important therapeutic target in the treatment of microvascular disorders.
Manea
Adrian
A
Nicolae Simionescu Institute of Cellular Biology and Pathology, 8, B.P. Hasdeu Street, Bucharest, Romania.
Raicu
Monica
M
Simionescu
Maya
M
D000804:Angiotensin II / Q000378:metabolism*; D000818:Animals; D002118:Calcium / Q000378:metabolism; D049109:Cell Proliferation; D005947:Glucose / Q000378:metabolism*; D006801:Humans; D008969:Molecular Sequence Data; D009243:NAD / Q000378:metabolism; D009247:NADH, NADPH Oxidoreductases / Q000235:genetics / Q000378:metabolism*; D009249:NADP / Q000378:metabolism; D019255:NADPH Oxidases; D020286:Pericytes / Q000201:enzymology*; D010586:Phagocytes / Q000201:enzymology*; D021122:Protein Subunits / Q000235:genetics / Q000378:metabolism*; D012333:RNA, Messenger / Q000378:metabolism; D051381:Rats; D017208:Rats, Wistar; D017382:Reactive Oxygen Species / Q000378:metabolism; D013481:Superoxides / Q000378:metabolism
D021122:Protein Subunits; D012333:RNA, Messenger; D017382:Reactive Oxygen Species; D009243:NAD; D013481:Superoxides; D000804:Angiotensin II; D009249:NADP; D009247:NADH, NADPH Oxidoreductases; D019255:NADPH Oxidases; D005947:Glucose; D002118:Calcium