title: Vitamin E deficiency accelerates nitrate tolerance via a decrease in cardiac P450 expression and increased oxidative stress.
abstract: Organic nitrates, such as nitroglycerin (NTG), have been used to relieve the symptoms of angina pectoris. However, their biochemical mechanisms of action, particularly in relation to the development of tolerance, are incompletely defined. It has been reported that supplemental antioxidants such as vitamin E attenuate the development of nitrate tolerance. Therefore, we examined the role of vitamin E in the regulation of nitrate tolerance. Continuous NTG infusion induced nitrate tolerance in rats after 48 h, and vitamin E concentrations decreased in a time-dependent manner in tissues and plasma. Vitamin E supplementation (0.5 g/kg diet) maintained higher concentrations of vitamin E during NTG infusion. The onset and extent of the tolerance, estimated by the decrease in blood pressure following NTG bolus injection during the infusion of NTG, were accentuated in the vitamin E-deficient group. Vitamin E supplementation inhibited nitrate tolerance 48 h after NTG infusion. Cardiac P450 expression (CYP1A2) assessed by immunoblotting, markedly decreased 48 h after NTG administration in control rats. The supplementation of vitamin E significantly attenuated the decrease in P450. Treatment of NTG enhanced vascular superoxide production (L-012 chemiluminescence, DHE fluorescence). The peak of lipid peroxidation and free radical generation in the heart was reached before tolerance developed. In contrast, vitamin E-deficient hearts had lower P450 expression and higher free radical generation than control hearts. To evaluate other vitamin E-inhibitable mechanisms of nitrate tolerance, we studied the NO-cGMP pathway. NTG markedly reduced the vasodilator-stimulated phosphoprotein (VASP) serine 239 phosphorylation (specific substrate of cGMP-activated protein kinase I; cGK-I) in tolerant hearts. Vitamin E inhibited the depletion of pVASP. In conclusion, because continuous NTG infusion causes vitamin E depletion as well as nitrate tolerance, vitamin E deficiency may further accelerate nitrate tolerance via an increase in oxidative stress, the reduced bioconversion because of decreased P450 expression, and impairment of the NO/cGMP pathway in tolerant heart tissues.
authors.affiliation: Department of Food and Health Science, Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama University, Shikata-cho, Japan. yukiko-m@md.okayama-u.ac.jp
authors.forename: Yukiko
authors.initials: Y
authors.lastname: Minamiyama
authors.forename: Shigekazu
authors.initials: S
authors.lastname: Takemura
authors.forename: Seikan
authors.initials: S
authors.lastname: Hai
authors.forename: Shigefumi
authors.initials: S
authors.lastname: Suehiro
authors.forename: Shigeru
authors.initials: S
authors.lastname: Okada
mesh_terms: D000818:Animals; D001011:Aorta / Q000737:chemistry / Q000378:metabolism; D015815:Cell Adhesion Molecules / Q000378:metabolism; D006152:Cyclic GMP / Q000378:metabolism; D019388:Cytochrome P-450 CYP1A2 / Q000378:metabolism*; D015536:Down-Regulation; D004361:Drug Tolerance; D015227:Lipid Peroxidation / Q000187:drug effects; D008297:Male; D008840:Microfilament Proteins / Q000378:metabolism; D009206:Myocardium / Q000737:chemistry / Q000166:cytology / Q000201:enzymology*; D009566:Nitrates / Q000008:administration & dosage; D009569:Nitric Oxide / Q000378:metabolism / Q000652:urine; D005996:Nitroglycerin / Q000008:administration & dosage*; D018384:Oxidative Stress; D010750:Phosphoproteins / Q000378:metabolism; D010766:Phosphorylation; D051381:Rats; D017382:Reactive Oxygen Species / Q000378:metabolism; D012694:Serine / Q000378:metabolism; D014665:Vasodilator Agents / Q000008:administration & dosage*; D014810:Vitamin E / Q000032:analysis / Q000097:blood; D014811:Vitamin E Deficiency / Q000378:metabolism*; D024502:alpha-Tocopherol / Q000008:administration & dosage; D000098662:Vasodilator-Stimulated Phosphoprotein
chemical_list: D015815:Cell Adhesion Molecules; D008840:Microfilament Proteins; D009566:Nitrates; D010750:Phosphoproteins; D017382:Reactive Oxygen Species; D014665:Vasodilator Agents; D000098662:Vasodilator-Stimulated Phosphoprotein; D014810:Vitamin E; D009569:Nitric Oxide; D012694:Serine; D019388:Cytochrome P-450 CYP1A2; D005996:Nitroglycerin; D006152:Cyclic GMP; D024502:alpha-Tocopherol