Lic syndrome. We have evidence that their combination produced synergistic improved vascular insulin resistance And metabolic diseases in this model of metabolic syndrome. Methods Animals. All operations were 5-alpha-reductase carried out in accordance with approved institutional guidelines for animal experimentation by the Committee on Animal Care and Use of Kumamoto University t. M Nnliche Wistar Kyoto rats, spontaneously hypertensive rats, and rats, SHRcp a rat model of metabolic syndrome, were purchased from Japan SLC. All rats have an animal with a 12 h light and darkness again U standard chow and water ad libitum, housed. Drugs. Candesartan, an ARB, was kindly provided by Takeda Pharmaceuticals available. Amlodipine, a CCB, was purchased from Wako Pure Chemical. N Ӭ Nigrol arginine methyl ester was purchased from Dojindo. The experimental protocol. Eleven weeks old SHRcp were divided into four groups and were orally administered vehicle, candesartan, candesartan or amlodipine and amlodipine for 4 weeks. Preliminary experiments showed that the dose of candesartan and amlodipine on Exerted similar hypotensive effect in SHRcp. Age-matched WKY and SHR rats were used as the controlled There were orally administered the vehicle and for 4 weeks. The K Body weight was measured periodically. The blood pressure and heart rate were measured by plethysmography before tail cuff and weeks 2, 3 and 4 after the start of the drug Measured sen treatment. After 4-w Weeks of treatment, all rats were bet with ether Exerted, and the heart, aorta, carotid artery, and subcutaneous and visceral adipose tissue were quickly excised for biochemical analysis and histology as described below in detail. Vascular ring preparation E and organ chamber experiments. Isometric tension studies were performed as previously briefly described.12 were cut the jugular vein of rats into 5 mm rings with special care to preserve the endothelium, and placed in Organb Of filled with modified Tyrode’s buffer with 95% O2 and 5% CO2 vented at 37.
The Pr Preparations were attached to a force transducer, and isometric tension was recorded on a polygraph. A resting tension of 1 g was f Maintained during the experiment. Vessel rings were primed with KCL and then precontracted with phenylephrine. After the plateau was reached, the rings increasing concentrations of acetylcholine, sodium nitroprusside or insulin to the cumulative concentration-response curves to obtain suspended. Measurement of vascular Ren superoxide. Thoracic aortas, removed from rats immediately were frozen in Tissue Tek embedding medium Dihydroethidium October was used to evaluate tissue levels of superoxide in situ as previously described.13 Briefly, fluorescence microscopy was Dihydroethidium visualized by fluorescence using an excitation length of 520 540 nm and an emission rhodamine filter length. Dihydroethidium tissue fluorescence was recorded with the same exposure time, and he was quantified in terms of values, the treated tissues TCR Pathway obtained from vehicle group. Western blot analysis of aortic tissue Eiwei and fat. The detailed method was previously described.14 shortly after aortic extracts proteins or fat Were subjected to gel electrophoresis on sodium dodecyl sulfate-polyacrylamide and transmission performance is subjected to polyvinylidene difluoride membrane.