Male Sprague Dawley rats will be housed in animal conditions by maintaining standard conditions of 25 degree Celsius room temperature, 12hour light/day cycle and 40-50% humidity.The rats will be provided with free access to food and water ad Libitum.The rats will be made unconscious using intra peritoneal injections of 80 mg/kg Ketamine and 10 mg/kg Xylazine. The anaesthesia will be sustained using supplementary injections.The procedure of tracheotomy will be performed followed by endotracheal intubation using a 16 gauge catheter. The entire procedure will be carried out on a heating pad to maintain the body temperature at 37 degree Celsius .
A rectal thermometer will be used to monitor the body temperature.Mechanical ventilation will be supplied to the rats by room air supplemented with oxygen such that a stroke volume of 1-1.5 ml/100g of rat weight and 52 strokes per minute. This will enable in sustaining the normal range of arterial blood pressure and blood pH. A catheter which is filled with 10IU/ml heparinised saline will be introduced into the right carotid artery.This will assist in collecting blood samples and measuring blood pressure at different times if follow up. ECG waves and arterial blood pressure will be assessed through data acquisition system.A cannula will be inserted into the tail vein to dispense Evans blue dye.
The heart of the rat is exposed by performing left thoracotomy between third and fourth inter coastal spaces.The incision made will be expanded using blunt ended retractors.The pericardial sac around the heart will be slit in order to identify the left descending coronary artery. After the determination of the position of left coronary descending artery, a 6-0 silk ligature will be passed around the artery a little distant from the origin site. The two ends if the ligature will be tied together.Partial closure of the chest will be executed.
The animals will be given a period of 30 minutes to recover from the effects during which heart relate and blood pressure will also become stabilised.After blood pressure and heart rate will become stable hemodynamic parameters will be measured for a time period of 15 minutes.The experimental protocol will then be initiated.