Study On The Expression Of KCa2.3/3.1 In Patients With Valvular Atrial Fibrillation And Thrombosis

Objectives:The purpose of this study was creating MS and MR porcine model to clarify the differences in expression and distribution of calcium-activated potassium channel KCa2.3/3,1 in different parts of the animal model,and to guide the selection of the most suitable part of human myocardial tissue specimens in further experiments;To comparing the pathological differences of myocardial tissue and the expression of KCa2.3/3.1 channel protein and related genes in myocardial tissue of patients with isolated mitral valve disease,valvular atrial fibrillation,valvular atrial fibrillation and thrombosis,and control group.To investigate the relationship between KCa2.3/3.1 channel and valvular atrial fibrillation and thrombosisMethods:1.The animal model of mitral valve disease in small southern pigs was established by using the method of suturing at the mitral valve anterior and posterior valve junctions and the method of mitral valve detachment before and after the mitral valve insufficiency model.5 cases in each case,4 cases in success,and 5 cases in control group(13 cases in all).After being bred for 10 months,they were sacrificed.Different myocardial tissue specimens(left atrium near pulmonary vein opening,left atrial appendage,atrial septum,right atrium)were taken.Papillary muscle,a total of 5 sites,13 cases of animals,a total of 65 specimens),after Western Bloting detection using SPSS 24.0 software for statistical analysis of the results,clear KCa2.3/3.1 two-channel protein in different parts of the expression of myocardial The differences in distribution guide the choice of the optimal material for the human myocardial tissue specimens in further experiments.2.Select 50 patients with rheumatic mitral valve disease admitted to the First Affiliated Hospital of Kunming Medical University and Fuwai Cardiovascular Hospital of Yunnan Province from May 2016 to October 2017,including simple mitral stenosis(MS),Simple mitral insufficiency(MR),mitral stenosis with atrial fibrillation(MS+AF),mitral insufficiency with atrial fibrillation(MR+AF),mitral stenosis with atrial fibrillation and thrombosis(MS+AF+ Thrombosis)10 patients each.A total of 10 patients with atrial septal defect,ventricular septal defect,and heart-occupying disease who had milder disease and did not involve hemodynamic changes related to mitral valve disease were selected as the control group.Left atrial appendage or atrial septal tissue specimens were taken as subjects prior to CPB(according to the results of the first part of the study,"KCa2.3/3.1 two channels showed no statistical difference in the left atrial appendage and atrial septum tissue").It is not suitable for patients with left atrial appendage to be replaced with atrial septum.)Paraffin sections were taken from specimens taken from human tissues,HE,Masson staining,and KCa2.3/3.1 channel-specific immunofluorescence staining.Qualitative comparisons were made between simple mitral valve disease(MS and MR)and valvular atrial fibrillation(MS+AF).Differences in myocardial tissue and differential expression of KCa2.3/3.1 channel protein between MR+AF),thromboembolic atrial fibrillation and thrombosis(MS+AF+thrombosis)patients and controls,and simultaneous detection by Western Blotiong and RT-PCR The expression of KCa2.3/3.1 channel protein and related gene KCNN3/4 in each sample was analyzed.SPSS 24.0 software was used for statistical analysis to quantitatively compare the expression differences between them.Results:1.Successfully established MS and MR animal models in 4 cases,a total of 8 cases(including 1 case in the MR group died in the modeling process,1 case died in the MS group during the postoperative observation),and the total module rate was 80%.2.Western Bloting results showed that the expression of KCa2.3/3.1 channel protein in the left atrium near the pulmonary vein was the highest in the myocardial specimens,which was significantly different from the other four sites(P<0.01);followed by the left atrial appendage and There was no significant difference between the two groups in the atrial septum(P>0.05),but they were significantly different from those in the right atrium and papillary muscle(P<0.01).The expression of KCa2.3/3.1 channel protein was lower in the right atrium and papillary muscle,MS,The right atrium was significantly higher in the MR group than in the papillary muscle(P<0.01).There was no significant difference between the right papillary muscle and the right atrium in the control group(P = 0.16>0.05).3.60 cases of patients with left atrial appendage(or atrial septum)were collected under the guidance of animal experimental results,including 10 cases of control,MS,MR,MS+AF,MR+AF,and MS+AF+thrombus groups.4.The results of HE and Masson staining of myocardial tissue in the patients showed that:compared with the control group,the mitral valve disease(MS and MR group)was thicker,looser,edema,and the nucleus was enlarged,there was vacuolization,and the connective tissue increased.Collagen fibers are rich and dense;in patients with valvular atrial fibrillation(especially with thrombosis),the above pathological manifestations are more prominent,with large and large vacuoles,and the nuclei are compressed and concentrated on the edges,and a large number of fibroplasias surround the myocardial cells,which are rich and dense..5.Immunohistochemical staining of myocardial tissue sections showed that the expression of KCa2.3/3.1 channel protein in myocardial tissue of patients with isolated mitral valve disease(MS and MR)was higher than that of control group;patients with VAF(MS+AF and In the MR+AF group,the expression of KCa2.3/3.1 channel protein in myocardial tissue was higher than that in patients with simple mitral valve disease(MS and MR),and even more complicated with thrombosis(MS+AF+thrombus).6.The results of Western Bloting and PCR showed that the expression of KCa2.3/3.1 channel protein and related genes in the myocardium of patients with mitral valve disease(MS and MR)was higher than that of the control group(P<0.05).;The expression of KCa2.3/3.1 channel protein and related genes in the myocardium of patients with valvular atrial fibrillation(MS+AF and MR+AF)was significantly higher than that of patients with simple mitral valve disease(MS and MR).(P<0.01),MS group was higher than MR group,MS + AF was higher than MR+AF group,the difference was statistically significant(P<0.05);Patients with thrombotic valvular atrial fibrillation(MS + AF + thrombus group)The expression of KCa2.3/3.1 channel protein and related genes in myocardium was significantly higher than that in valvular atrial fibrillation patients w:ithout thrombosis(MS+AF and MR+AF)(P<0.05).Conclusions:1.Establishing reliable and stable MS and MR animal models by using the method of suturing at the mitral valve anterior and posterior valve junctions and the mitral valve chordal detachment method to accurately simulate abnormal hemodynamics in MS and MR patients.Learn to change.2.The expression and distribution of KCa2.3/3'1 channels in different parts of the myocardium are different,roughly as follows:left atrial proximal pulmonary vein opening>left atrial appendage>atrial septum>right atrium>papillary muscles:most of the distribution of left atrial proximal pulmonary veins,The absolute advantage;followed by the left atrial appendage and atrial septum,there was no statistical difference between the two;the distribution of KCa2.3/3.1 channels in the right atrium and papillary muscle is less,but the right atrium is slightly more than the papillary muscle,the difference is Statistical significance.The differential expression of KCa2.3/3.1 channels in different parts of the myocardium is associated with mitral valve disease,but not with mitral valve disease(stenosis or insufficiency).3.Mitral valve disease caused by myocardial cell disorder,edema,steatosis(vacuole),irregular nuclear morphology,fibrous connective tissue proliferation and other myocardial injury and fibrosis characteristics.In patients with VAF,especially in patients with VAF and thrombosis,these pathological changes were further aggravated,with irregular cell morphology and arrangement,marked edema,large amounts of steatotic vacuoles,large proliferation of collagen fibers,and accumulation of nuclei on the edges of cells.The normal myocardial tissue structure disappears.4.KCa2.3/3.1 two-channel protein and its related gene KCNN3/4 in mitral valve disease in patients with elevated myocardial expression,associated with mitral valve disease,and related to the type of mitral valve disease,high mitral stenosis Mitral valve insufficiency.5,KCa2.3/3.1 two-channel protein and its related gene KCNN3/4 in patients with valvular atrial fibrillation increased expression,and atrial fibrillation-related,valvular atrial fibrillation than simple mitral valve disease.6,KCa2.3/3.1 two-channel protein and its related gene KCNN3/4 in patients with valvular atrial fibrillation and thrombosis in the expression of elevated myocardial,and thrombus-related,thrombotic valve atrial fibrillation than non-thrombotic valve Atrial fibrillation.