动物造模,巨细胞病毒性心肌炎模型 z-bio 2023-09-27 830

　　Human cytomegalovirus herpesvirus β The subfamily is a linear double stranded DNA virus with latent activation biological characteristics. HCMV infection is quite common in China, and it is mostly asymptomatic and latent in the immune normal population. However, in some special populations (including fetuses, infants, and immunosuppressants), HCMV infection can lead to serious diseases. Giant cell myocarditis is the deadliest form of myocarditis, characterized by rapid deterioration of the condition and poorer prognosis compared to other forms of myocarditis. Establishing a mouse model of cytomegalovirus myocarditis can provide a good research platform for the treatment, drug screening, disease pathogenesis, and disease prognosis evaluation of cytomegalovirus myocarditis.

　　1. Pathogens were taken from healthy pure BALB/c mice (female, 6-8 weeks old, weighing 20-22g) and injected intramuscularly with methylprednisolone injection at a dose of 2mg per mouse, once every 4 days. On the second day after the first intramuscular injection of methylprednisolone, the mice were intraperitoneally inoculated with 10 000 pfu MCMV K181 virus suspension. On the 14th day after the virus injection, the mice were euthanized by enucleation, Sterile isolation of mouse salivary glands was carried out by placing them in serum-free Dubecco MEM Eagle culture medium (DMEM, Gibco company). Each salivary gland was homogenized with 1ml of serum-free DMEM culture medium, centrifuged at 3000r/min for 30 minutes, and the virus supernatant was collected and stored in a refrigerator at -80 ℃ for future use.

　　2. Animal health pure 5-week-old BALB/c mice.

　　Intraperitoneal inoculation of cytomegalovirus MCMV K181 virus suspension (10000 pfu per animal) through 3 infection routes.

　　4. Pathological manifestations: On the 3rd day after intraperitoneal inoculation of 10000 pfu virus suspension in mice, the myocardial tissue homogenate of the mice was taken, and 200 supernatants were centrifuged μ Inoculated with MEF, obvious cytopathic CPE (cell enlargement, rounding, and shedding) can be observed on the 7th day. On the 3rd day after MCMV infection, inflammatory lesions mainly infiltrating neutrophils and monocytes were found in the myocardial interstitium and vascular endothelium of mice, reaching a peak at 7-10 days. Inflammatory cell infiltration was observed in the myocardial interstitium, including myocardial structural dissolution, destruction, and vacuolar degeneration. Subsequently, inflammatory cell infiltration gradually decreased (Figure 32-3). After 3 to 4 months of virus infection, inflammatory cell infiltration lesions in the myocardial interstitium of mice can still be observed.

　　5. Advantages and disadvantages: Cytomegalovirus belongs to the herpesviridae family and has strict species specificity, so animal models of human cytomegalovirus (HCMV) infection cannot be established. By infecting mice with mouse cytomegalovirus, a suitable mouse model of cytomegalovirus induced myocarditis can be established for in vivo simulation research. The myocardial tissue damage caused by mouse cytomegalovirus and HCMV infection is also very similar. The target organs infected by mouse cytomegalovirus include liver, lung, kidney, salivary gland, and heart, which can cause corresponding tissue pathological damage and can detect mouse cytomegalovirus. The pathogenicity of cytomegalovirus is weak, and the K181 strain of mouse cytomegalovirus