Research The Expression Of HCN2Gene MRNA In Atrial Fibrillation Patients With Mitral Stenosis Of Rheumatic Heare Disease

Objective: To analysis the expressions of HCN2mRNA in atrialfibrillation patients and sinus rhythm patients in mitral stenosis of rheumaticheart disease,and explore the possible adjustment mechanism of atrial fibrillationwith mitral valve stenosis of rheumatic heart disease,then aim to lay thetheoretical foundation that HCN2play an important role in atrial fibrillation ofRHD through the mRNA level.Methods:1. There were43patients those mitral stenosis of rheumaticheart disease but without other disease,and their NYHA level were betweenⅡtoⅢwhose come from the department of cardiothoracic surgery in the Firstafiliated hospital of Guangxi medical university.We divided the patients into twogroups thruogh the results of the colour sonography and the cardiogram:theconrtol group were26atrial fibrillation patients with mitral stenosis ofrheumatic heart disease(AF group),and the matched group were17sinusrhythm patients with mitral stenosis of rheumatic heart disease(SR group).2. RT-PCR method weas used to analyze the difference expressions of HCN2 in the atrial fibrillation patients with mitral stenosis of rheumatic heart diseaseand in the sinus rhythm patients with mitral stenosis of rheumatic heart diseaseby the ratio of HCN2gene expression and the β-actin mRNA expression forevaluating the mRNA expression level of HCN2gene.3. Through the application of fluorescent dye SYBR Premix Ex TaqTMⅡ，themethod of real time fluores-cence quantitative PCR(FQ-PCR) was set up todetected the difference expressions of HCN2in the atrial fibrillation patientswith mitral stenosis of rheumatic heart disease and in the sinus rhythm patientswith mitral stenosis of rheumatic heart disease by the standard of2-△△CT.4.All the date were expressed by x±s and the inspection level was0.05,the measurement date was analysis by T test through the SPSS16.0.Results:1.By the analysis of the RT-PCR method, The levels ofexpressions of HCN2mRNA in the atrial fibrillation patients in mitral stenosisof rheumatic heart disease and in the sinus rhythm patients with mitral stenosisof rheumatic heart disease were0.99±0.18and0.28±0.06, the differenceexpressions in the two groups was significant(t=10.39，P＜0.01),and the resultaim that theHCN2mRNA was high expression in the atrial fibrillation patientswith mitral stenosis of rheumatic heart disease.2.The standard curve of HCN2and β-actin were set up, then the resultindicates the amplification efficiency and the correlation coefficent of HCN2and β-actin were E%=113.75, r2=0.997, E%=111.89and r2=0.995. That pointout HCN2and β-actin have the same amplification efficiency, then explain thatthis FQ-PCRmethod was dependable.3.The FQ-PCR result reveal that the levels expressions of HCN2mRNA in the atrial fibrillation patients with mitral stenosis of rheumatic heart diseaseand in the sinus rhythm patients with mitral stenosis of rheumatic heart diseasewere0.70±0.14and0.39±0.08, the result aim that not only the differenceexpressions in the two groups was significant(t=3.31，P＜0.01), but also theHCN2mRNA was high expression in the atrial fibrillation patients with mitralstenosis of rheumatic heart disease.Conclusion:1. The HCN2mRNA was high expression in the atrialfibrillation patients with mitral stenosis of rheumatic heart disease,this may bethe cause that it add up the hyperpolarization-activated cation current(If)， thenit resulting in atrial fibrillation.2. The levels expressions of HCN2mRNA in the atrial fibrillationpatients with mitral stenosis of rheumatic heart disease were higher than thesinus rhythm patients with mitral stenosis of rheumatic heart disease through thePCR method.This result reveal that the highe-levels expressions of HCN2mRNA were play a important role in the process of the atrial fibrillation,and itmay be the possible adjustment mechanism of atrial fibrillation in rheumaticheart disease.