Research On The Pathogenesis Of Atrial Fibrillation By Interference On Cx40 And Kv1.5 Potassium Channel Protein

OBJECTIVEExploring the methods for primary culture of atrial myocytes and the expression of connexin40(Cx40)and Kv1.5 channel proteins on patients with atrial fibrillation(AF),which can investigate the relationship between connexin 40 and Kv1.5.Methods1.The left atrial appendages of atrial fibrillation patients(mitral valve replacement and MAZE surgery)and sinus rhythm patients were treated by type I collagenase digestion to culture the atrial myocytes.Then the primary atrial myocytes were identified through immunohistochemistry by type II Collagen antibody and troponin antibody simultaneously.2.The expression of Kv1.5 mRNA and protein were detected by PCR and Western-blot respectively through RNA interference on Cx40 mRNA.RESULTS1.Under the microscope,a few circular cells attached the culture flask after 24hours' primary culture.72 hours later,most of them attached the culture flask.The cells will cover the bottom in the next three days.It is suggested that the primary cells which identified by immunohistochemistry were atrial myocytes.2.Compared with the control group,the m RNA and protein expression of Cx40 and Kv1.5 in atrial myocytes were significantly decreased in atrial fibrillation patients.After the RNA interference on Cx40,the expression of Cx40 was decreased,as well as the Kv1.5 mRNA and protein.CONCLUSION1.The atrial myocytes of patients with atrial fibrillation can be cultured by type I collagenase digestion.2.The interference on Cx40 of atrial fibrillation patients can significantly reduce the expression of Kv1.5 mRNA and protein.