| Heart failure(HF)is a group of complex clinical syndromes caused by abnormal changes in cardiac structure and/or function due to a variety of causes,resulting in dysfunction of ventricular contraction and/or diastolic function.Myocardial remodeling is the main pathophysiological process of HF occurrence and development,as well as an important factor of disease development.In recent years the incidence of HF has a rising trend in our country,the case fatality rate and hospitalization rate is still in the high level again,rising as the population ages,coronary heart disease,diabetes,high blood pressure,obesity and other risk factors of heart failure disease incidence is rising,HF disease control and prevention and treatment in our country still faces huge challenges,and a serious threat to the survival of patients with HF and life quality.Although there is clear evidence that drugs are effective in the treatment of systolic heart failure,the effect of drugs on delaying,stopping or even reversing myocardial remodeling has not been conclusively demonstrated,and the five-year survival rate for heart failure patients is still less than 50%.Therefore,it is of great significance to explore the intervention effect of TCM on the occurrence and development of heart failure.Qishentaohong granules,as a clinical protocol prescription,were confirmed by prospective randomized controlled studies to improve TCM syndrome score,Ejection fraction(EF),n-terminal b-type natriuretic peptide(nt-probnp),6-minute walking distance(6MWD),quality of life,and 6-month re-hospitalization rate in heart failure patients.Before the onset of heart failure,myocardial remodeling generally occurs first,including changes in heart structure,shape and function caused by pathophysiological stimulation.As time goes on,chronic stress factors or disease factors will cause ventricular dilatation,decreased ventricular contractility and eventually develop into HF.It can be said that myocardial remodeling is the most important pathophysiological process in the occurrence and development of HF.Pilot study confirmed that stilbene and peach particles mainly composed of astragalus,codonopsis,salvia miltiorrhiza can decrease pressure load leads to heart failure animal model of left ventricular chamber wall thickness,left ventricular mass index,improve the E/A ratio and left ventricular ejection fraction,lower left ventricular end-diastolic pressure,can inhibit myocardial remodeling,and at the same time improve the ventricular diastolic and systolic function.Therefore,correction of myocardial remodeling may be one of the mechanisms of qishentaohong granules to intervene heart failure.PurposeBy intervening in the mouse model of heart failure caused by constricting thoracic aorta,and based on mTOR autophagy related signaling pathway,the effect of qishantaohong granules on myocardial remodeling in the mouse model of heart failure was investigated.MethodsThe mouse heart failure model was established by narrowing the thoracic aorta,and the experimental animals were randomly divided into model group,sham operation group,Chinese medicine group(qishengtaohong granules),western medicine group(enalapril maleate),Chinese medicine+rapamycin group,and Chinese medicine+3-methyl adenine group.Two dynamic observations were made at the time points of modeling,2 weeks and 4 weeks after intervention.At the overall level,the general conditions of mice,cardiac function monitoring by small animal ultrasound,degree of myocardial hypertrophy,and heart weight and lung weight(standardized by body weight)after death were observed.At the tissue level,a complete cross section of the heart was taken for routine HE and Masson staining to observe the hypertrophy of cardiomyocytes and the proliferation of collagen fibers in extracellular matrix.At the cellular level:autophagosome formation was observed under transmission electron microscopy,and apoptosis level was detected by TUNEL method.At the gene molecular level:left ventricular myocardial tissue of mice in the sham operation group,model group and traditional Chinese medicine group were selected for smallRNA sequencing.According to the sequencing results,the key smallRNA of autophagy was selected,and the expression level and difference of target smallRNA in the myocardial tissue of animals in each group were analyzed by QPCR to evaluate whether the expression trend was consistent with the sequencing results.Finally,the effects of qishantaohong granules on myocardial remodeling in mice with heart failure were discussed.Results:1.General conditions and survival conditions:mice in the model group and the intervention group showed obvious symptoms of heart failure after surgery,while mice in the sham operation group showed no obvious abnormalities after recovery.Mice died in all groups except the sham group.2.Body weight and standardized cardiopulmonary weight:in terms of body weight,2 weeks after the operation,compared with the sham operation group,the traditional Chinese medicine group increased,showing a statistical difference(P<0.05).Compared with the model group,there was no statistical difference in each group.4 weeks after surgery,compared with the control group and model group,traditional Chinese medicine,Chinese medicine joint rapamycin group,traditional Chinese medicine combined 3 methyl adenine and group were increased,with statistical difference(P<0.05),by the same group before and after comparison,found that enalaprilat group 4 weeks after surgery after two weeks of a higher weight,statistically significant(P<0.05).In terms of standard cardiac weight,compared with the sham group,the model group and enalapril group significantly increased 2 weeks after surgery(P<0.05),and there was no statistical difference between the model group and the model group.Four weeks after surgery,compared with the sham group,the model group significantly increased(P<0.05),compared with the model group,the traditional Chinese medicine group,the traditional Chinese medicine combined with rapamycin group,and the enalapril group significantly decreased(P<0.05).In terms of standardized lung weight,compared with the sham group,the model group and enalapril group significantly increased(P<0.05)2 weeks after surgery,but there was no statistical difference between the model group and the model group.Four weeks after surgery,compared with the sham group,the model group significantly increased(P<0.05),compared with the model group,the traditional Chinese medicine group,the traditional Chinese medicine combined with rapamycin group,and the enalapril group significantly decreased(P<0.05).Two weeks after the operation,the model group was significantly higher than the sham group(P<0.05),and there was no statistical difference between the other groups and the sham group.Compared with the model group,the TCM group,the TCM combined with rapamycin group,the TCM combined with 3-methyl adenine group,and the enalapril group were all significantly lower(P<0.05).Four weeks after the operation,there was no statistical difference between the sham operation group and the model group.Compared with the sham operation group and the model group,the group of TCM combined with 3-methyl adenine decreased(P<0.05),while the group of TCM combined with rapamycin increased(P<0.05).3.Evaluation of cardiac function:Two weeks after surgery,(1)left ventricular ejection fraction(LVEF%),shortening fraction(FS%)and left ventricular end-diastolic dimension(left ventricular end-diastolic dimension)were decreased in the model group compared with the sham group(P<0.05).LVIDd)and diastolic left ventricular posterior wall thickness(LVPWd)increased during diastolic period(P<0.05).In the intervention group,FS,LVEF and LVIDd decreased to different degrees in the TCM group,the TCM combined with rapamycin group,the TCM combined with 3-methyl adenine group,the enalapril group(P<0.05),and the LVIDd increased(P<0.05).LVPWd increased in traditional Chinese medicine group and enalapril group(P<0.05).(2)compared with the model group,LVEF and FS increased in the TCM group(P<0.05),LVIDd increased in the TCM combined with 3-methyl adenine group(P<0.05),and LVPWd decreased in the TCM group,the TCM combined with rapamycin group,and the TCM combined with 3-methyl adenine group.There were statistically significant differences in LVEF values in the TCM group,the TCM combined with rapamycin group,the TCM combined with 3-methyl adenine group and the enalapril group.Four weeks after surgery,compared with the sham group,FS and LVEF values in the model group decreased(P<0.05)and LVIDd and LVPWd increased(P<0.05).In the intervention group,the FS and LVEF values decreased in the TCM combined with rapamycin group,the TCM combined with 3-methyl adenine group,the enalapril group,and the LVPWd values increased(P<0.05),and the LVIDd values increased in the TCM group,the TCM combined with rapamycin group,the TCM combined with 3-methyl adenine group,and the enalapril group(P<0.05).(2)compared with the model group,FS and LVEF values increased in the TCM group,the TCM combined with rapamycin group,the TCM combined with 3-methyl adenine group,and the enalapril group,while LVPWd values decreased(P<0.05).The LVIDd value of the TCM group,the TCM combined with rapamycin group and the enalapril group decreased(P<0.05).(3)comparison between groups and before and after the same group showed that FS and LVEF values in the traditional Chinese medicine combined with rapamycin group and enalapril group were higher than before,while FS and LVEF values in the model group were lower than before(P<0.05).Four weeks after surgery,the LVEF value of the TCM combined with rapamycin group was higher than that of the TCM combined with 3-methyl adenine group(P<0.05),and that of the enalapril group was higher than that of the TCM combined with 3-methyl adenine group(P<0.05).The LVIDd value of the traditional Chinese medicine combined with rapamycin group,enalapril group and model group was higher than that of the model group(P<0.05).(4)although there was no statistical difference between some groups,comparative analysis of the mean indicated that the traditional Chinese medicine group had more advantages than the other intervention groups in restoring the ability of heart contraction.4.Left ventricular myocardial histopathologic evaluation:1)the control:is the group mice heart cross section geometry,no obvious abnormal myocardial tissue morphology,orderly arrangement of myocardial fibers,horizontal grain clarity,endometrial cells and myocardial continuity is good,basic cells in the middle nucleus,patina uniform coloring,cell morphology,size,did not see cell edema and inflammatory cell infiltration,myocardial fibrosis cells did not see.2)model group:compared with the sham operation,the cross-section of the heart in this group showed cardiac hypertrophy,increased heart diameter,and changed geometric shape.Hypertrophy and necrosis of cardiomyocytes were observed,and thickening of blood vessel wall and infiltration of inflammatory cells were observed in the peripheral blood vessels.The fibrosis of aizen was seen in Masson's staining and was more severe 4 weeks after surgery than 2 weeks after surgery.3)Chinese medicine group:compared with model group,the group left ventricular myocardial hypertrophy,to a lesser degree,neat arrangement of myocardial cell and visible compensatory hypertrophy,cardiac geometric shape change,peripheral vascular wall thickening,inflammatory cells infiltration,Masson staining aizen part significantly lighter than model group,prompt myocardial fibrosis,to a lesser degree,and the degree of fibrosis,inflammatory cells infiltration situation at 4 weeks after surgery,is better than 2 weeks after surgery.4)TCM+3-methyl adenine group:compared with the TCM group,there was no obvious pathological improvement in this group,showing ventricular wall hypertrophy,heart geometry change,thick peripheral vascular wall,inflammatory cell infiltration,and 2 weeks after surgery,myocardial cell swelling and inflammatory cell infiltration were worse than 4 weeks after surgery.Masson staining showed that the degree of myocardial fibrosis in this group was lighter than that in the TCM+rapamycin group,and the degree of fibrosis at 4 weeks after surgery was heavier than that at 2 weeks after surgery.5)TCM+rapamycin group:compared with the TCM group,no significant pathological improvement was observed in this group,showing ventricular wall hypertrophy,heart geometry changes,peripheral vascular wall thickening,and inflammatory cell infiltration.Masson staining indicated partial areas of myocardial fibrosis.6)enalapril group:compared with the model group,the pathological conditions of this group were improved,including ventricular wall hypertrophy,heart geometry changes,peripheral vascular wall thickening,inflammatory cell infiltration,and orderly arrangement of cardiomyocytes.Masson staining indicated that the myocardial fibers in this group were lighter than those in the model group.5.Observation of autophagosome and other ultrastructure of left ventricular myocardial tissue by transmission electron microscopy:Were compared with control group,model group,TCM group,Enalapril group,TCM+RA group,TCM+3-MA autophagy bubble,autophagy and autophagy-lysosome populations,visible with a crescent or goblet,double layer or multilayer membrane characteristics,has a trend of the cytoplasm components of autophagy,with double layer or multilayer membrane vacuole structure,which contain cytoplasm,such as mitochondria,endoplasmic reticulum,and ribosome of autophagy and the monolayer film,the wrapped slurry composition has the degradation of autophagy-lysosome,disordered arrangement of muscle cells gradually,mitochondrial sparse loosely arranged,irregular,visible mitochondria structure was destroyed.Generally,4 weeks after surgery was more serious than 2 weeks after surgery,and the arrangement and integrity of mitochondria and muscle cells were the worst in TCM+3-ma group.6.Possible molecular mechanisms based on second-generation sequencing and bioinformatics analysis:According to the miRNA results of the differences between the traditional Chinese medicine group and the model group at 2 and 4 weeks after surgery(the difference multiple was greater than 1,P<0.05),it can be found that the differences of miRNA in the traditional Chinese medicine group were mir-200a-3p,mir-200c-3p,mir-708-3p,mir-200b-3p,mir-6715-5p,mir-96-5p,mir-7689-3p,mir-1983,and mir-382-5p at 4 weeks after surgery.The differences of mirnas include mir-7225-5p,mir-671-5p,mir-677-5p,mir-582-3p,mir-208b-3p,mir-lOla-5p,mir-1844-3p,and mir-18 1 b-2-3p.Gene Ontology enrichment analysis showed that a total of 123 terms were enriched 2 weeks after surgery and 215 terms were enriched 4 weeks after surgery,suggesting a close relationship with autophagy.Hierarchical analysis of KEGG differentially expressed gene pathways revealed that at 4 weeks after surgery,the top 20 most differentially expressed pathways included the mTOR and Lysosome pathways,which are closely related to autophagy.Conclusion:1.qishentaohong granules can effectively improve the cardiac function and myocardial remodeling of pressure-loaded heart failure mice.2.in the compensatory stage of heart failure,qishen peach red granules up-regulated the autophagy of cardiomyocytes to play a protective role,and down-regulated the autophagy of cardiomyocytes in the decompensated stage of heart failure to reduce the damage effect of excessive autophagy on cardiomyocytes3.based on the mTOR signaling pathway,through the regulation of related miRNA,the bidirectional regulation of the autophagy level of cardiomyocytes to exert the role of autophagy to protect the myocardium is one of the mechanisms for the intervention of qishen peach red granules in HFrEF and myocardial remodeling. |
PDF Full Text Request