Effects Of Shipi Decoction On Cardiac Function And Cardiomyocyte Apoptosis In Rats With Chronic Heart Failure

ObjectiveTo investigate the effects of Shi Pi Yin on cardiac function,NT-proBNP,Caspase-3 and myocardial cell apoptosis in rats with chronic heart failure(CHF)induced by Isoproterenol(ISO).MethodsHealthy male SD rats(n=80)weighing 180-200g were randomly divided into normal control group(n=10)and model group(n=70)after adaptive feeding for 7 days.The model group was injected with ISO[5mg/(kg·d)]intraperitoneally for 14 days.Will building successful CHF rats were randomly divided into model control group(n=40)[n=10,saline 7.5ml/(kg·d)],Shi Pi Yin low dose group(n=10,Shi Pi Yin mixed suspension lavage 3.75 g(student)/(kg·d)],Shi Pi Yin medium dose group(n=10,Shi Pi Yin mixed suspension lavage 7.5 g(student)/(kg·d)],Shi Pi Yin high dose group(n=10,Shi Pi Yin mixed suspension lavage 15g(student)/(kg·d)],after 14d continuous intervention,The LVIDs,LVIDd,LVEF and FS were measured by color doppler ultrasound.Serum NT-proBNP content was determined by ELISA;The expression of Caspase-3 protein in each group was detected by immunohistochemistry and its expression level was analyzed.TUNEL staining was used to detect the apoptosis of rat myocardial tissue and to calculate the apoptosis index.All data were processed by SPSS 22.0 statistical software,and the measurement data were expressed as mean±standard deviation((?)±s),and P<0.05 was considered statistically significant.Results1.General observation of rats in each group:Rats in the normal control group ate and drank normally during the experiment,their fur was shiny,and their mental status and activity level were good.Twenty-one rats died during modeling,with a mortality rate of 30%.The rats in the model group gradually began to huddle together,and the amount of food,water and activity was less than that of the control group before and after the model building,and the coat gloss was less than that before,and the scrotum was edema.During the intervention period,the spirit of the rats in each dose group gradually improved,the amount of drinking and drinking water and activity increased,and the symptoms of scrotal edema improved.2.Echocardiographic indexes of rats in each group:Compared with the normal control group,LVIDs in the model control group and in the low,medium and high dose groups were significantly increased,and LVEF and FS were significantly decreased(P<0.05 or P<0.01).LVIDd increased in both the model control group and the low,medium and high dose groups,but the difference was not statistically significant(P>0.05).Compared with the model control group,LVEF and FS were significantly increased in the low,medium and high dose groups(P<0.05 or P<0.01).LVIDs and LVIDd decreased in the low,medium and high dose groups compared with the model control group,but the difference was not statistically significant(P>0.05).Comparison between the two groups showed that the increase of LVEF and FS was the most obvious in the medium dose group(P<0.01).There was no significant difference in LVEF and FS levels between the two groups(P>0.05).3.Serum NT-proBNP content of rats in each group:Compared with the normal control group,the serum NT-proBNP level in the model control group and the low,medium and high dose groups of Shi Pi Yin increased significantly(P<0.01).Compared with the model control group,the serum level of NT-proBNP was significantly reduced in the low,medium and high dose groups(P<0.01).In the comparison between the two groups,the level of NT-proBNP in the medium dose group decreased most significantly(P<0.05).There was no significant difference in serum NT-proBNP content between the two groups(P>0.05).4.Expression of Caspase-3 protein in rat cardiomyocytes in each group:A small amount of brown particles were observed in the cytoplasm of rat cardiomyocytes in the normal control group,while large amounts of brown particles were observed in the model control group,and brown particles of different degrees were observed in the low,medium and high groups.The analysis of the expression level of Caspase-3 immunopositive products showed that the average optical density of Caspase-3 in the model control group and the low,medium and high dose groups of Shi Pi Yin all increased compared with the normal control group(P<0.01).Compared with the model control group,the average optical density of Caspase-3 in the low,medium and high dose groups decreased(P<0.01).The average optical density of Caspase-3 decreased most significantly in the medium dose group(P<0.05 or P<0.01).There was no significant difference between the two groups(P>0.05).5.Apoptosis rate of rat cardiomyocytes in each group:The nuclei of rat cardiomyocytes in the normal control group were mostly blue,and a small amount of nuclei were brown;the nuclei of model control group were significantly increased,and different degrees of brown nuclei were observed in each group.The calculated apoptosis rate of cardiomyocytes showed that compared with the normal control group,the apoptosis rate of cardiomyocytes in the model control group and the low-dose,medium-dose and high-dose groups were significantly increased(P<0.01).Compared with the model control group,the apoptosis rate of myocardium cells in the low,medium and high dose groups decreased(P<0.01).The apoptosis rate of myocardial cells in the medium dose group was the most significantly decreased(P<0.01).There was no significant difference between the low-dose and high-dose groups(P>0.05).ConclusionShi Pi Yin can improve cardiac function in CHF rats to a certain extent,reduce serum NT-proBNP,inhibit myocardial cell apoptosis,and thus delay the development of heart failure.The mechanism of action may be related to the down-regulation of Caspase-3 protein expression.