The rabbit isolated aortic strip assay can be used to observe the effects of drugs on the diastolic and contractile functions of vascular smooth muscle. Observation of the effects of drugs on aortic strips treated by hyperkalemic depolarization and norepinephrine allows a preliminary analysis of the site of action of the drug on vascular smooth muscle cells: whether it is through voltage-dependent calcium channels or receptor-manipulated calcium channels.
Operation method
specimen preparation method
Principle
The rabbit isolated aortic strip assay can be used to observe the effects of drugs on the diastolic and contractile functions of vascular smooth muscle. Observation of the effects of drugs on aortic strips treated by hyperkalemic depolarization and norepinephrine allows a preliminary analysis of the site of action of the drug on vascular smooth muscle cells: whether it is through voltage-dependent calcium channels or receptor-manipulated calcium channels.
Materials and Instruments
Rabbit Move 1. Preparation of rabbit aortic strips. A rabbit, hit the head to stun, quickly dissect the chest, separate the thoracic aorta, in the proximal and distal end of the diaphragm cut, quickly placed in oxygenated Kirschner's solution, remove the extravascular connective tissue and fat, wash off the clot, gently set in the same thickness as the glass rod, and then with ophthalmic scissors will be aorta sheared into a width of 3 ~ 4 mm, the length of the spiral strip of 2 ~ 3 cm. Both ends were threaded and ligated, and placed in the Mackenzie bath, one end of which was connected to the L-shaped tube in the water bath, and the other end was connected to the tension transducer, with the preload of the tension transducer being 2 g. The temperature of the water bath was adjusted to 37 ± 1 °C. The water bath was filled with 30% of the volume of the aorta, and the temperature was adjusted to 30% of the volume of the aorta. The temperature of the water bath was adjusted to 37±1 °C. The Mackenzie bath was filled with 30 ml of Kirschner's solution and a gas mixture of 95% O2 and 5% CO2 was passed through the tube. Connect the recorder to the tension transducer and adjust the zero point. Caveat 1. the preparation of aortic strip specimens should be gentle, do not pull hard, so as not to damage the smooth muscle tissue of the aorta. 2. The Kirschner's solution must be freshly prepared on a temporary basis. 3. When introducing the gas mixture into the bath tube, pay attention to the ventilation volume, usually 40-60 bubbles/minute. For more product details, please visit Aladdin Scientific website.
Potassium Chloride Ketamine Solution Prazosin Norepinephrine
Powerlab Tension Transducer Rabbit Table Surgical Instruments Gauze Cotton Balls Thermostatic Bath Bath Tubs Syringe 4-Gauge Needle 5-Gauge Needle
2. After the aortic strip has stabilized for approximately 1 hour, the normal tension curve is traced and the drugs are administered in the following order:
(1) Add 3×10-3 mol/L noradrenaline 0.1 ml and, after a maximal response, flush the specimen.
(2) After recovery of the aortic strip, add 10-2 mol/L prazosine 0.2 ml and repeat (1) after 15 minutes. Allow the effect to stabilize and rinse the specimen.
(3) Add 4 mol/L potassium chloride 0.1 ml to the tube and rinse after the effect is evident.
(4) After the curve is stabilized, add 10-2 mol/L prazosine 0.2 ml and repeat (3) after 15 minutes.