Enzyme And Virus Titers Changes In The Mouse Myocardial Cells Infected With After Coxsackie Virus In Vitro

Object: coxsackie virus B group (CVB) is the first pathogen which causeviral myocarditis in human, especially CVB3. Viral myocarditis is acommonly infectious disease in children, and in adults is sporadic, whichcauses fever, sudden, acute heart failure and dilated cardiomyopathy. Thepathogenesis mechanisms have not been completely understood and now itis believed mostly that the direct injury of virus and the immune,inflammatory response caused by virus infection may be responseful for thedisease. There is no effective preventive measures and specific drug therapyavailable in clinical at present. It is very imperative to explore the process andthe changes of the body after infecting CVB3. So it is needed to study on thechanges in myocardial cells after infecting CVB3to providing the basis for thestudy of the pathogenesis of viral myocarditis and development process.Methods：1Culture of murine cardiomyocytes: the heart of mice were tokensterilely from C57BL/6mice and shredded. The cells were collected aftertrypsin and collagenase digestion and seeded to6-well plates and cultured inthe CO2incubator at37C. anti-B-actin，anti-vimertin antibodies were used tocardiomyocyte identification.2Infection of myocardial cells with Coxsackie virus: the myocardial cellswere infected with coxsachie virus at80%confluent. The CPE was observeddaily and the supernatant were collected at1day,2days,3days,4days,6days,8days,10days post infection and stored at-30C.3Enzyme detection: the supernatants collected after the virus infectionwere measured for the activities of troponin I (cTnI), high-sensitivityC-reactive protein (CRP), creatine kinase (CK), lactate dehydrogenase (LDH),creatine kinase (CK-MB), α hydroxybutyrate dehydro-genase (α-HBDH), myoglobin (MYO), homocysteine (HCY).4virus titration: the virus teters of the supernatants were measured inHela cells.Results:1Cultured myocardial cells in mice grow well, immuohistochemistrystaining showed anti-B-actin positive and anti-vimertin negative, which wasthe cherecteristc of the myocardial cells2Enzymes of supernatants at different time points after the infection ofmice cardiomyocytes with coxsackievirus were measured, the results showedon the third day the enzymes reached its peak.3The virus production of the supernatants at different time points weretitered in Hela cells. The results showed that: at the first three days there werevirus produced and then the coxsackievirus titer declined, which indicated thecells may be not the sensitive cells for the virus.Conclusion: CVB3is a major pathogen of viral myocarditis. Myocardialcells infected with the CVB3will destroy the skeleton structure, releasing avariety of enzymes in the early stages and cause directly induce apoptosis, butCVB3could not proliferate very well in myocardial cell. Myocarditis inducedby CVB3may be immune damage.