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1Department of Biomedical Sciences, 2Dalton Cardiovascular Research Center, and 3Departments of Medical Pharmacology and Physiology and Internal Medicine, University of Missouri, Columbia, Missouri 65211; and 4Department of Kinesiology, University of Waterloo, Waterloo, Ontario, Canada N2L 3G1
Submitted 22 January 2003 ; accepted in final form 3 March 2003
Our purpose was to determine the effects of gender and exercise training on endothelial nitric oxide synthase (eNOS) and superoxide dismutase (SOD) protein content of porcine skeletal muscle arteries and to evaluate the role of 17
-estradiol (E2) in these effects. We measured eNOS and SOD content with immunoblots and immunohistochemistry in femoral and brachial arteries of trained and sedentary male and female pigs and measured estrogen receptor (ER) mRNA and 
-ER and -ER protein in aortas of male and female pigs. Results indicate that female arteries contain more eNOS than male arteries and that exercise training increases eNOS content independent of gender. Male and female pigs expressed similar levels of -ER mRNA and protein and similar amounts -ER protein in their arteries. E2
concentrations as measured by RIA were 180 ± 34 pg/ml in male sera and 
5 pg/ml in female sera, and neither was changed by training. However, bioassay indicated that biologically active estrogen equivalent to only 35 ± 5 pg/ml was present in male sera. E2 in female pigs, whether measured by RIA or bioassay, was 24 pg/ml at peak estrous and 2 pg/ml on day 5 diestrus. The free fraction of E2 in sera did not explain the low measurements, relative to RIA, of E2. We conclude that 1) gender has significant influence on eNOS and SOD content of porcine skeletal muscle arteries; 2) the effects of gender and exercise training vary among arteries of different anatomic origin; 3) male sera contains compounds that cause RIA to overestimate circulating estrogenic activity; and 4) relative to human men, the male pig is not biologically estrogenized by high levels of E2
reported by RIA, whereas in female pigs E2 levels are lower than in the blood of human women.
skeletal muscle blood flow; vascular smooth muscle; nitric oxide; estradiol-free fraction; estrogen; radioimmunoassay; biological activity; endothelial nitric oxide synthase
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