Use of flavonoid compound originated from Dracocephalum moldavica in protecting myocardial ischemia reperfusion injury
An anti-myocardial ischemia and reperfusion injury technology, applied in the anti-myocardial ischemia-reperfusion injury effect of luteolin, kaempferol, luteolin field
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Embodiment 1
[0018] Protecting isolated rat myocardium from ischemia-reperfusion injury
[0019] 1. Langendoff device assembly Assemble the Langendoff device, set the pressure height to 80cm; start the super constant temperature water bath, connect the heat preservation circulating water, and keep the constant temperature at 37°C; fill in the Lorraine's solution, open the oxygen valve, fill in an appropriate amount of oxygen, and pass The liquid medicine pump pumps the Lorraine's solution into the Langendoff device, checks the connection, adjusts and maintains a stable flow rate, and exhausts the perfusion air bubbles. Connect the tee, connect the balloon at one end and the transducer at the other end, connect to the computer through the converter, open the BL-420F biological function experiment system, click input signal → channel 1 → left ventricular pressure, and click the Excel mark to record data.
[0020] 2. The isolated heart ischemia / reperfusion model was replicated in rats and w...
Embodiment 2
[0039]Protection against myocardial ischemia-reperfusion injury in rats
[0040] 1. Ischemia-reperfusion injury model preparation 280-320g male SD rats were lightly anesthetized with 20% urethane in the abdominal cavity, intubated through the oral cavity, and ventilated by a ventilator with positive pressure (respiratory frequency 80 times / min, tidal volume 20mL, respiratory ratio 1: 2, air pressure 0). Insert the needle electrode into the skin of the three limbs, connect to channel 1 or 2 of the BL-420s biological function experiment system, open the system software and the corresponding channel, and measure the ECG. The hair in the middle of the left chest was shaved and disinfected. The skin was cut longitudinally 1 cm at the place where the heart beat, and the muscle layer was bluntly separated between the 4th and 5th intercostals, and the intercostal muscles and pleura were stretched to enter the chest cavity. The chest cavity was pulled open, the pericardium was torn, ...
Embodiment 3
[0059] Protect cardiomyocytes from hypoxia / reoxygenation injury
[0060] 1. Primary cardiomyocyte culture Take suckling mice, separate the heart under sterile conditions, wash with cold PBS, and cut into 1mm pieces 3 The left and right pieces were digested in batches with 0.0625 g / L trypsin at 37°C under constant temperature and low-speed stirring, 8 min each time, 4 times in total. After each supernatant was collected, the digestion was terminated with an equal volume of DMEM medium containing 15% FBS, and then centrifuged (1000rpm×10min), the cardiomyocytes were collected and washed once with the medium, and then cultured for 90min with differential attachment . Then the cell suspension was at a density of 5×10 5 cells / mL were inoculated in 96-well culture plate, at 37°C, 5% CO 2 Under the condition of culture, the culture medium was changed every 2 days.
[0061] 2. Primary cardiomyocytes were grouped for drug pretreatment and hypoxia / reoxygenation injury model prepar...
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