PMC:509282 / 12343-15337 JSONTXT

{"target":"https://pubannotation.org/docs/sourcedb/PMC/sourceid/509282","sourcedb":"PMC","sourceid":"509282","source_url":"https://www.ncbi.nlm.nih.gov/pmc/509282","text":"Tissue Preparation\nMale Sprague-Dawley rats (central animal facility, National Institute of Pharmaceutical Education and Research (NIPER), India), 160–200 g, were kept in controlled environmental conditions with room temperature 22 ± 2°C, humidity 55 ± 5% and 12-h light/dark cycles. All the animals had free access to food and water. The rats were divided into two dietary groups and fed with standard rat normal pellet diet (NPD) (3.8 kcal/g, carbohydrate 67%, protein 21%, fat 12% kcal) and HFD (5.3 kcal/g, carbohydrate 17%, protein 25%, fat 58% kcal). Composition of HFD is described in Table 1.\nTable 1 Composition of HFD\n(g)\nPowdered pellet diet 364\nLard 310\nCasein 250\nCholesterol 10\nDL-Methionine 3\nYee-sac powder 1\nVitamin and mineral mix powder 60\nSodium chloride 2\n\nBiochemical measurements\nBlood samples from the retro orbital plexus of anaesthetized rats (Pentobarbitone 45 mg/kg, i.p.,) were collected into the heparinized tubes and immediately centrifuged at 5000 rpm for the separation of plasma. Plasma was stored at -20°C until assayed. The plasma was used for the estimation of glucose (Qualigens, Mumbai, India), triglycerides, and cholesterol (Chema diagnostica, Jesi, Italy) by commercial kits. Plasma insulin was determined by radioimmuno assay using rat insulin as standard (Linco research, St. Charles, MO, USA)\n\nIntra peritoneal glucose tolerance test (IPGTT)\nGlucose tolerance tests were carried out after four weeks of feeding of both NPD and HFD. After an overnight fast, blood samples were collected from the retro orbital plexus. Glucose levels were measured at time zero (0 min) and glucose was injected into the rats (2 g/Kg/4 ml, i.p.,). Additional blood samples were taken at 15, 30, 60 and 120 min. following the glucose load. Plasma glucose levels were measured by the glucose oxidase reaction (GOD/POD) using commercial kit (Qualigens, Mumbai, India). Area under the curve (AUC) was calculated for both NPD and HFD fed rats.\n\nVascular Studies\nAfter 4 weeks of feeding, rats were sacrificed by cervical dislocation. The section of the aorta from between the aortic arch and the diaphragm was removed from the euthanized rats and placed in oxygenated, modified Krebs-Henseleit solution (KHS; in mM: 118 NaCl, 4.7 KCl, 25 NaHCO3, 2.6 CaCl2 2H2O 1.2 NaH2PO4, 1.2 MgCl2 6H2O, 5.5 glucose). With the help of a steel rod, aortic endothelium was deliberately denuded. The aorta was cut into helical strips 3 mm wide, 20 mm long and then placed in a well-oxygenated (95% O2-5% CO2) bath of 10-ml KHS with one end connected to a tissue holder and the other to an isotonic transducer (Bio Devices, Ambala, India). The tissue was equilibrated for 60 min under a resting tension of 1.0 g. At the beginning of each experiment, aortic strips were primed with depolarizing concentration (90 mM) potassium chloride (KCl). After the equilibration period, contractile responses to various concentrations of 5-HT (10 nM-30 μM) and Ang II (1 nM-300 nM) were recorded.","divisions":[{"label":"title","span":{"begin":0,"end":18}},{"label":"p","span":{"begin":19,"end":600}},{"label":"table-wrap","span":{"begin":601,"end":785}},{"label":"label","span":{"begin":601,"end":608}},{"label":"caption","span":{"begin":610,"end":628}},{"label":"p","span":{"begin":610,"end":628}},{"label":"table","span":{"begin":629,"end":785}},{"label":"tr","span":{"begin":629,"end":632}},{"label":"td","span":{"begin":629,"end":632}},{"label":"tr","span":{"begin":633,"end":658}},{"label":"td","span":{"begin":633,"end":653}},{"label":"td","span":{"begin":655,"end":658}},{"label":"tr","span":{"begin":659,"end":668}},{"label":"td","span":{"begin":659,"end":663}},{"label":"td","span":{"begin":665,"end":668}},{"label":"tr","span":{"begin":669,"end":680}},{"label":"td","span":{"begin":669,"end":675}},{"label":"td","span":{"begin":677,"end":680}},{"label":"tr","span":{"begin":681,"end":696}},{"label":"td","span":{"begin":681,"end":692}},{"label":"td","span":{"begin":694,"end":696}},{"label":"tr","span":{"begin":697,"end":713}},{"label":"td","span":{"begin":697,"end":710}},{"label":"td","span":{"begin":712,"end":713}},{"label":"tr","span":{"begin":714,"end":731}},{"label":"td","span":{"begin":714,"end":728}},{"label":"td","span":{"begin":730,"end":731}},{"label":"tr","span":{"begin":732,"end":766}},{"label":"td","span":{"begin":732,"end":762}},{"label":"td","span":{"begin":764,"end":766}},{"label":"tr","span":{"begin":767,"end":785}},{"label":"td","span":{"begin":767,"end":782}},{"label":"td","span":{"begin":784,"end":785}},{"label":"sec","span":{"begin":787,"end":1346}},{"label":"title","span":{"begin":787,"end":811}},{"label":"p","span":{"begin":812,"end":1346}},{"label":"sec","span":{"begin":1348,"end":1972}},{"label":"title","span":{"begin":1348,"end":1395}},{"label":"p","span":{"begin":1396,"end":1972}},{"label":"title","span":{"begin":1974,"end":1990}}],"tracks":[]}