Study On Mechanism Of Cardiac Developmental Regulatory Proteins Interacting With Influenza Virus H1N1

Congenital heart disease(CHD)that has being threat to human healthy seriously has become one of the highest mortality of human birth defects disease.Wherein 8-10%of familial congenital heart disease and the rests displayed distributed congenital heart disease.The genetic and environmental factors,in which environmental factors include intrauterine infections,drugs,and environmental pollution,and the interactions between two all of them are the important reason for the occurrence of CHD.Influenza virus as one of the important factors of intrauterine infection has been concerned hotly in recent years.It has been reported that influenza virus infection could significantly increase the risk of cardiac malformations occur.Thus,studies on the pathogenesis of congenital heart disease have being made great progress.However,the reason how influenza virus and other environmental factors cause disease incidence of CHD molecular mechanism is unclear on researches at home and abroad.It has been reported that H1N1 influenza virus proteins can interact with a variety of physiological proteins of the host cell.These interactions bring about passive effect on the expression of the host cell proteins and viral proteins themselves in the level of processing and translation after transcription.To study the relationship between the H1N1 flu virus protein and heart development regulatory protein,the study based on our previous digital gene repression profiling sequencing filter and the bioinformatics screening analysis,screened to two important candidate genes,the Dvl2 and the Gata6.And virus protein fragments were recombinant,researched the interaction relationship between the H1N1 flu virus protein and heart development regulatoryprotein,we hypothesis the H1N1 flu virus can bind directly to their promoter.This article,transiently transfected the influenza virus recombinant expression plasmid that with flag-tagged into H9C2 cell,the expression showed that the H1N1 influenza virus can enter the host cell nucleus;further using the chromatin immunoprecipitation(CHIP)to study the relations between the influenza virus and the regulatory protein promoter,and the results showed that the H1N1 influenza virus can be successfully incorporated into the promoter regions of the Dv12 and the Gata6 of transcription factor;Finally,tested for the influenza virus proteins and two different fragments candidate binding protein promoter was provided,showed that the promoter of the Dv12 and the Gata6 mainly concentrated on non-structural proteins NS1 of H1N1 influenza virus.The Gata6 that is a key regulator of factor of cardiac early development can not only regulate myocardial cells occurs,differentiation,and cardiac precursor cells to form linear heart tube,but also bring about human cardiac malformations due to mutation.Therefore,the latter parts of this experiment focused on exploring the transcriptional regulation on the NS1 protein exert effect on the different lengths of the Gata6 promoters.The experiment results shows thar in 48 hours of transfection NS 1 was significant activation of its upstream 945bp fragments,but inhibitor for its transcription was founded in the sixth day.In this research,the influenza virus recombinant plasmids were constructed,the Dvl2 and the Gata6,two important genes,related to heart development were selected,we explored the mechanism of interaction between influenza virus and these two genes at the cellular level.We research for further study how the H1N1 influenza virus causes and in turn mediated molecular mechanisms of congenital heart disease laying the foundation,for the study of the relationship between congenital heart disease and other environmental factors provides a useful reference.