The Study On The Effect And Mechanism Of Qiling Tongguan Decoction On Ventricular Remodeling In Patients With Heart Failure

Heart failure(HF)is a series of syndromes in cardiovascular diseases which develop to the end stage.Patients have severe illness,poor prognosis and high mortality.Clarifying the pathogenesis of heart failure is especially important for the prevention and treatment of heart failure.Ventricular remodeling(VR)is the most basic pathogenesis of heart failure and the critical factor causing the deterioration of cardiac function.Ventricular remodeling include many complex pathophysiological processes,such as inflammation,cardiomyocyte hypertrophy,cardiomyocyte death,myocardial fibrosis,neuroendocrine regulation,molecular signal transduction pathway and so on.Preventing and delaying ventricular remodeling are the key to improve clinical symptoms and cardiac function in patients with heart failure.Traditional Chinese medicine has unique advantages in improving ventricular remodeling in heart failure,but the mechanism is not clear.The objective of this study is to explore the effect and molecular mechanism of Qiling Tongguan Decoction on ventricular remodeling through clinical and animal experiments,which further clarify the effectiveness and scientificity of traditional Chinese medicine in the treatment of heart failure.Experiment 1:Clinical effect of Qiling Tongguan Decoction on patients with heart failure and its effect on ventricular remodelingObjectiveTo observe the clinical effect of Qiling Tongguan Decoction on patients with chronic heart failure and its effect on myocardial fibrosis and cardiomyocyte apoptosis.Methods1.Patients with qi deficiency and blood stasis were randomly divided into western medicine group and traditional Chinese medicine combined with Western medicine group.The western medicine group was treated with conventional western medicine,and the traditional Chinese medicine combined with Western medicine group was treated with western medicine combined with Qiling Tongguan Decoction for 3 months;2.The main indexes included:①Transthoracic echocardiography;②TCM symptom efficacy,TCM single symptom score and TCM total symptom score;③Blood biochemical indexes:BNP,sST2,GDF-15;④Cardiac injury markers:CK-MB,MYO,cTnT;⑤Inflammatory factors:TNF-α,IL-6,IL-1β;⑥The relative content of BCL-2,BAX and caspase-3 mRNA in peripheral blood mononuclear cells.Results1.General situation:During the follow-up and treatment,82 patients who finally completed in the study,42 cases in the western medicine group and 40 cases in the traditional Chinese medicine combined with Western medicine group.There were no significant difference between the two groups in age,gender,height,weight,systolic blood pressure,diastolic blood pressure,smoking history,drinking history,basic diseases(Hypertension,Diabetes,Hyperlipidemia),and medication(ACEI/ARB,β-Blockers,Duretics,Aldosterone receptors antagonist,Asprin and Statin).2.Cardiac ultrasound markers:After treatment,EF and FS of patients in the two groups were significantly higher than that before treatment(P<0.01);The levels of EF and FS in traditional Chinese medicine combined with western medicine group were higher than that in western medicine group(P<0.05);3.TCM symptom score:After treatment,the scores of palpitation,fatigue,shortness of breath,wheezing,fear of cold,cold limbs,chest tightness and pain,purple lips,infracostal ruff,exposure of neck tendons and edema of lower limbs in the two groups were lower than those before treatment(P<0.01);The total score of TCM syndrome in traditional Chinese medicine combined with western medicine group was significantly lower than that in western medicine group(P<0.01);The curative effect of TCM syndrome showed that the total effective rate of Western medicine group was 80.95%,and that of Western medicine combined with traditional Chinese medicine group was 87.5%;4.Blood biochemical indexes:The levels of BNP,sST2 and GDF-15 in the two groups after treatment were significantly lower than that before treatment(P<0.01);The levels of BNP and sST2 in the traditional Chinese medicine combined with western medicine group were lower than that in the western medicine group(P<0.01),and there was no significant difference in the level of GDF-15 between the two groups(P>0.05);5.Myocardial markers:The levels of CK-MB and cTnT in the two groups after treatment were significantly lower than that before treatment(P<0.01),MYO was not different than that before treatment(P>0.05),and CK-MB and cTnT in the western medicine combined with TCM group were lower than that in the western medicine group after treatment(P<0.05),and there was no significant difference in the level of MYO between the two groups(P>0.05);6.Inflammatory factors markers:After treatment,IL-6,IL-1β and TNF-α in the two groups were significantly lower compared with that before treatment(P<0.01).IL-6,IL-1βand TNF-α levels in the traditional Chinese medicine combined with western medicine group were significantly lower than that in western medicine group(P<0.01);7.qRT-PCR:There were significant differences in the relative expression of BCL-2,BAX and caspase-3 mRNA between the two groups before and after treatment(P<0.01);The relative expression of BCL-2 in traditional Chinese medicine combined with western medicine group was higher than that in western medicine group(P<0.01),and the relative expression of BAX and caspase-3 were lower than that in western medicine group(P<0.05,P<0.01);8.Safety indexes:There was no significant difference in ALT,AST,Urea and Cr between western medicine group and traditional Chinese medicine combined with western medicine group before and after treatment(P>0.05).Conclusions1.Qiling Tongguan Decoction combined with western medicine may improve cardiac function and TCM syndrome in patients and effectively reduce myocardial cell injury,myocardial fibrosis,cardiomyocyte apoptosis and improve ventricular remodeling during heart failure;2.The results of peripheral blood mononuclear cells show that Qiling Tongguan Decoction may reduce cardiomyocyte apoptosis by blocking mitochondrial apoptosis pathway.Experiment 2 Based on CaMKⅡδ/PLB/SERCA2a signaling pathway study on the effect and mechanism of Qiling Tongguan Decoction on ventricular remodeling in mice with heart failureObjectiveBased on the previous clinical trials,CaMKⅡδ/PLB/SERCA2a signaling pathway as a key role to further explore the effect of Qiling Tongguan Decoction on mouse model of heart failure,which to provide an objective and reliable scientific basis for the treatment of heart failure with traditional Chinese medicine.Methods1.C57BL/6J mice model of heart failure was prepared by abdominal subcutaneous injection of isoproterenol and were randomly divided into Control group(Control group),Model group(Iso group),Metoprolol group(MT group),Traditional Chinese medicine group(ZY group);2.The body weight was weighed before and after the experiment,the performance of mouse heart failure and the mortality were observed;the mice were tested by cardiac ultrasound;the mice were dissected and body weight,whole heart weight,left ventricular weight,lung weight were weighed and tibial length was measured;The heart tissue was dissected and divided into three parts:apex,heart and bottom.Hematoxylin-eosin staining was used to detect cardiomyocytes morphology and Sirius red staining was used to detect myocardial interstitial collagen fibers;The apex and bottom of the heart were Put into liquid nitrogen and frozen at-80℃ and the cardiac protein were extracted for Western blotting.Results1.General conditions of mice:During the study,there was no death in contorl group,and the mortality was 0%;Two rats died in Iso group,with a mortality rate of 20%;Three rats died in MT group,with a mortality rate of 30%;Two rats died in ZY group,with a mortality rate of 20%;Before treatment,HW,LVW and LUW were significantly higher in Iso group,MT group and ZY group(P<0.01);After drug intervention,HW,LVW and LUW were significantly lower in MT group and ZY group(P<0.01),and HW/BW,LUW/BW,HW/TL and LVW/TL were significantly lower(P<0.01)in ZY group.HW/BW and LUW/BW were significantly lower in ZY group than MT group(P<0.05,P<0.01);2.Cardiac ultrasound of mice:Compared with contorl group,LVAWd,LVIDs,LVPWd in Iso group increased significantly(P<0.01),EF and FS decreased significantly(P<0.01);Compared with Iso group,LVAWd,LVPWd,LVIDs of ZY group decreased(P<0.01),EF and FS increased significantly(P<0.01);3.HE staining and Sirius red staining of mice:There were obvious degeneration and necrosis of cardiomyocytes,infiltration of interstitial lymphocytes and proliferation of fibrous tissue in Iso group;The morphological changes of cardiomyocytes were improved and myocardial fibrosis was alleviated in MT group and ZY group after drug intervention;4.Western blot of mouse heart:In Iso group,the expression of CaMKⅡδ and PLB increased(P<0.01),the expression of SERCA2a and p-PLB decreased(P<0.01);After drug intervention,the expression of CaMKIIδ and PLB decreased(P<0.05,P<0.01)and the expression of SERCA2a and p-PLB increased(P<0.01,P<0.05);The expression of SERCA2a,CaMKIIδ,PLB,p-PLB were no significant difference between MT group and ZY group(P>0.05).Conclusions1.Qiling Tongguan Decoction can effectively protect against ventricular hypertrophy,cardiomyocyte degeneration and necrosis,myocardial fibrosis.2.Qiling Tongguan Decoction can regulate CaMKⅡδ/PLB/SERCA2a signaling pathway through inhibits downstream molecule CaMKIIδ and upstream the expression of SERCA2a and PLB phosphorylation.It can alleviate mitochondrion calcium overload,maintain calcium circulation homeostasis,enhance cardiac systolic and diastolic function and improve ventricular remodeling.