动物造模,心气虚细胞模型 z-bio 2023-10-16 816

　　(1) Reproduction method: Myocardial cell culture was performed on NIH mouse suckling mice 2-4 days after birth. The hearts of the suckling mice were taken and washed in Hank's solution. The atria were cut off and repeatedly rinsed to remove residual blood accumulation. The ventricles of the suckling mice were placed on the sidewall of a centrifuge tube, with each ventricle cut twice vertically and horizontally. 10ml of 0.1% trypsin was added and digested by vibration in a 37 ℃ water bath for 10 minutes. After natural precipitation, the supernatant was discarded and 10ml of trypsin was added, Place it in a 37 ℃ water bath for vibration digestion for 10 minutes, blow the tissue with a straw for 1 minute, and allow it to settle naturally. Transfer the supernatant to another sterile centrifuge tube, add 2ml of culture medium to terminate digestion, and continue to digest the sediment with trypsin according to the previous method. Collect the supernatant according to the previous method, repeat this process 6 times. Centrifuge the supernatant with a centrifuge at 500r/min for 5 minutes continuously. After discarding the supernatant, add the culture medium and repeat centrifugation once. Place the cell suspension in a culture bottle and let it stand for 1.5 hours in a 37 ℃ constant temperature incubator. Suck out the cell suspension, count it, and inoculate it onto a 24 well culture plate. Inoculate 2.5 myocardial cells per well × 100000 pieces, add culture medium (80ml of 1640 culture medium and 20ml of calf serum per 100ml) to 1ml culture plate per well, seal with transparent glue for 4 weeks, and culture in a 37 ℃ constant temperature incubator. After 2 days, change the culture medium once. After 3 days of cultivation, a monolayer of myocardial cells was taken for testing. The culture plate was placed in a container and filled with 99.99% high-purity nitrogen gas for 1 minute before sealing the container. The container was opened after 1, 2, and 3 hours of hypoxia, and oxygen (95% O2, 5% CO2) was filled in for 1 minute. After standing at 37 ℃ for 30 minutes, the cell supernatant was collected for biochemical testing, including lactate dehydrogenase (LDH), creatine kinase (CK), malondialdehyde (MDA), superoxide dismutase (SOD), etc.

　　(2) The characteristics of the model include a significant increase in LDH, CK, and MDA in the supernatant of cultured cardiomyocytes after 1, 2, and 3 hours of hypoxia, while a decrease in SOD activity. The most significant changes were observed when the hypoxia time was 3 hours.

　　(3) Patients with heart qi deficiency syndrome in comparative medicine often have clinical manifestations of heart dysfunction, such as palpitations, shortness of breath, and particularly severe movement. Myocardial ischemia can often lead to damage to heart function, leading to symptoms similar to heart qi deficiency syndrome such as palpitations, shortness of breath, and particularly severe movement. The damage to cardiac function caused by myocardial ischemia is ultimately caused by myocardial hypoxia caused by ischemia. From a theoretical analysis, the damage to heart function caused by myocardial ischemia and hypoxia should be considered as an important reason for the occurrence of heart qi deficiency syndrome. Using hypoxia as a simulated cause of heart qi deficiency syndrome, a heart qi deficiency cell model was established by culturing myocardial cells with hypoxia followed by oxygen injury. A new exploration was made in the modeling of heart qi deficiency syndrome. The author believes that. The formulation of the cell model of heart qi deficiency is worth discussing. According to the concept of traditional Chinese medicine syndromes, we can only say the corresponding changes in the expression of heart qi deficiency syndrome at the cellular level, so it seems inappropriate to determine it as the cell model of heart qi deficiency.