Establishment Of The Animal Model Of Pulmonary Hypertension In Congenital Heart Disease And The Expression Of MKL1

ExperimentⅠ Establishment of animal model of pulmonary hypertension in congenital heart disease at high altitudeObjective: The animal model of pulmonary hypertension induced by congenital heart disease was established by dividing the abdominal aorta and inferior vena cava fistula of SD rats.The animals were housed in hypoxic chamber to simulate the plateau hypobaric hypoxia.Methods: Male SD rats were in accordance with the inclusion criteria of a total of 40 cases,divided into four groups,Three groups were randomly selected underwent peritoneal shunt surgery,According to the altitude is divided into low altitude group(1000 meters),in the elevation group(3000 meters),high altitude group(5000 meters),another group as naive control group.Each group recruit 10 rats.Three operation groups recieved shunt surgery of abdominal aorta and inferior vena cava under high power microscope.Arteriovenous anastomotic incision for 2 to 3 mm.The rats underwent longitudinal cutting abdominal aorta and inferior vena cava to suture incision and communicated,after the completion of the operation in accordance with the altitude of feeding around in normoxia and in a hypobaric chamber for four weeks.The naive control group was not operated and feed around in normoxia chamber for four weeks.We observed the changes of body weight and the formation of pulmonary hypertension in rats by right heart catheterization.The right ventricular pressure,right ventricular free wall(RV),Left ventricle and interventricular septum(LV + S)were assessed in all groups,and the Fulton index(RV / LV+S)weight ratio was calculated as well.Results: The weight gain of the rats in the high altitude group,the middle altitude group and the low altitude group was significantly lower than that in the normal control group.Four groups of animals were calculated right ventricular free wall(RV),left ventricle and septum(LV + S),Fulton index(RV / LV +S),Pulmonary artery pressure.The results were statistically significant difference(P < 0.05).Conclusions: The animal model of abdominal aorta and inferior vena cava through the SD rats to form fistula formation of shunt congenital pulmonary hypertension,and to simulate different altitudes in hypobaric chamber in animal hypobaric hypoxia state,can provide an animal model for the mechanism of pulmonary hypertension in congenital heart disease and the mechanism of the impact of different altitudes on its severity.ExperimentⅡ Expression of MKL-1 in rat lung tissueObjective: At present,studies have shown that MKL-1(megakaryocytic leukemia factor 1)is the formation of pulmonary hypertension in one of the influence factors,the process of the formation of pulmonary hypertension in hypoxia,can regulate smooth muscle cell proliferation and change the structure and remodeling of pulmonary vessels.In this study,MKL-1 m RNA and protein expression were detected in four groups of lung tissues,and the mechanism of plateau pulmonary hypertension was initially studied from the level of protein and gene.The effects of different altitude on the degree of pulmonary hypertension were compared.Methods: The four groups of rats were killed after,take out a small amount of lung tissue,protein extraction,electrophoresis of protein quantitative valuation method,then SDS-polyacrylamide gel electrophoresis,transfer,closed(block),incubated in anti,washed,incubated for two resistant,washing,color(HRP enzyme),and finally comparison of MKL1 results in the four groups,and then use the PCR denaturation,annealing and extension of MKL1 gene amplification,the results were compared between the four groups.Results: The protein expression and gene expression of MKL1 in the normal control group,low altitude group,the moderate elevation group and the high altitude group were increased in turn,Compared with the normal control group,the difference was statistically significant(P < 0.05).Conclusions: Different altitudes can significantly affect the severity of plateau of congenital heart disease with pulmonary hypertension,as sea level rises,pulmonary hypertension is rising,in which MKL1 is involved in the formation of pulmonary hypertension,and the severity of the reaction in a certain degree of pulmonary hypertension.