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Role of Cu,Zn-SOD in the synthesis of endogenous vasodilator hydrogen peroxide during reactive hyperemia in mouse mesenteric microcirculation in vivo

¹Department of Medical Engineering and Systems Cardiology, Kawasaki Medical School, Kurashiki, Japan; ²Department of Cardiovascular Medicine, Tohoku University Graduate School of Medicine, Sendai, Japan; ³Department of Anesthesiology, Kyushu University Graduate School of Medical Sciences, Fukuoka, Japan; ⁴Department of Physiology, Tokai University School of Medicine, Isehara, Japan; and ⁵Department of Cardiac Physiology, National Cardiovascular Center Research Institute, Suita, Japan

Submitted 4 September 2007 ; accepted in final form 12 November 2007

We have recently demonstrated that endothelium-derived hydrogen peroxide (H₂O₂) is an endothelium-derived hyperpolarizing factor and that endothelial Cu/Zn-superoxide dismutase (SOD) plays an important role in the synthesis of endogenous H₂O₂ in both animals and humans. We examined whether SOD plays a role in the synthesis of endogenous H₂O₂ during in vivo reactive hyperemia (RH), an important regulatory mechanism. Mesenteric arterioles from wild-type and Cu,Zn-SOD^–/– mice were continuously observed by a pencil-type charge-coupled device (CCD) intravital microscope during RH (reperfusion after 20 and 60 s of mesenteric artery occlusion) in the cyclooxygenase blockade under the following four conditions: control, catalase alone, N^G-monomethyl-L-arginine (L-NMMA) alone, and L-NMMA + catalase. Vasodilatation during RH was significantly decreased by catalase or L-NMMA alone and was almost completely inhibited by L-NMMA + catalase in wild-type mice, whereas it was inhibited by L-NMMA and L-NMMA + catalase in the Cu,Zn-SOD^–/– mice. RH-induced increase in blood flow after L-NMMA was significantly increased in the wild-type mice, whereas it was significantly reduced in the Cu,Zn-SOD^–/– mice. In mesenteric arterioles of the Cu,Zn-SOD^–/– mice, Tempol, an SOD mimetic, significantly increased the ACh-induced vasodilatation, and the enhancing effect of Tempol was decreased by catalase. Vascular H₂O₂ production by fluorescent microscopy in mesenteric arterioles after RH was significantly increased in response to ACh in wild-type mice but markedly impaired in Cu,Zn-SOD^–/– mice. Endothelial Cu,Zn-SOD plays an important role in the synthesis of endogenous H₂O₂ that contributes to RH in mouse mesenteric smaller arterioles.

nitric oxide; endothelium-derived hyperpolarizing factor; arteriole; vasodilatation

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