| Background:Heart failure(HF)is the final stage and serious manifestation of the development of various cardiovascular diseases.Although the drug treatment methods for HF are constantly updated and improved,the treatment effect is still insufficient due to the existence of strict contraindications and adverse reactions.The efficacy of traditional Chinese medicine in treating HF has been unanimously recognized by scholars.Qishentaohong granules is an effective Chinese medicine prescription for treating HF.It has been proved to have a good therapeutic effect on HF in long-term clinical practice and previous studies,but the specific mechanism still needs further study.Therefore,this study explores the clinical efficacy,inflammatory response,and safety of Qishentaohong granules in patients with chronic HF in the clinical study.In terms of experimental research,the study first analyze the main components and its pharmacological effects of Qishentaohong granules,and further explore the immune regulatory mechanism of Qishen taohong granules in treating HF.Objective:1.To explore the effect of Qishentaohong granules on the clinical efficacy,inflammatory reaction and safety of chronic HF.2.To analyze the main components and its pharmacological effects of Qishentaohong granules.3.To explore the immunoregulatory mechanism of Qishentaohong granules in the treatment of HF.Study 1.Randomized cross-controlled trial of Qishentaohong granules in the treatment of chronic heart failure.Methods:This study is a prospective,randomized,single-blind,crossover trial.The patients with chronic HF who meet the inclusion criteria were randomly divided into two groups(group A and group B)in equal proportion.Treatment plan 1 was basic treatment+Qishentaohong granules,and treatment plan 2 was basic treatment+Chinese medicine placebo.Group A received treatment plan 1 in the first stage(P1),and treatment plan 2 in the second stage(P2).Group B was the opposite of Group A.The intervention period of P1 and P2 was 4 weeks respectively,and the washout period was 2 weeks.The primary outcome was left ventricular ejection fraction(LVEF).The secondary outcomes were New York Heart Association(NYHA)classification,left ventricular fractional shortening(LVFS),left ventricular end-diastolic diameter(LVIDd),left ventricular end-systolic diameter(LVIDs),left ventricular end-systolic diameter(LVIDs),TCM syndrome and symptom score,six-minute walk test(6MWT)and chronic heart failure integrated traditional Chinese and western medicine survival scale(CHFQLS).The exploratory outcomes were interleukin(IL)1β,IL-6,IL-10 and tumor necrosis factor(TNF)-α.The safety outcomes were blood routine,urine routine,liver and kidney function and adverse reactions.The above outcomes were evaluated before and after P1 treatment,and before and after P2 treatment.Results:A total of 58 patients with chronic HF were included in this study,including 29 patients in group A and 29 patients in group B.During the study period,3 cases were lost in group A and 3 cases in group B.Finally,26 subjects in group A and 26 subjects in group B completed the trial.1.The primary outcome;LVEF:there was difference before and after treatment in treatment plan 1(P<0.05),there was no difference before and after treatment in treatment plan 2(P>0.05),and there was no difference between groups after treatment(P>0.05).2.The secondary outcomes:(1)LVFS,LVIDs and LVIDd:there were differences before and after treatment in treatment plan 1(P<0.05),there were no differences before and after treatment in treatment plan 2(P>0.05),and there were no differences between groups after treatment(P>0.05).(2)NYHA classification:P1:the total effective rate of group A was 80.76%,and that of group B was 57.69%,there was a difference between the two groups(P<0.05).P2:the total effective rate of group A was 50.00%,and that of group B was 84.62%,there was difference between the two groups(P<0.05).(3)6MWT:there were differences before and after treatment in treatment plans 1 and 2(P<0.05),and there were differences between groups after treatment(P<0.05).(4)TCM syndromes and symptoms:in terms of TCM syndromes,shortness of breath,fatigue,asthma,blood stasis and facial limb edema,there were differences between treatment plans 1 and 2 before and after treatment(P<0.05),and there were differences between groups after treatment(P<0.05).In terms of palpitation,chest tightness(pain)and abdominal distension,there were differences between the two treatment plans before and after treatment(P<0.05),but there were no differences between the two groups after treatment(P>0.05).(5)CHFQLS:in terms of total score,physiological function,role restriction,vitality and total score,there were differences before and after treatment in treatment plan 1 and 2(P<0.05),and there were differences between groups after treatment(P<0.05).In terms of social function and medical support,there were differences between the two treatment plans before and after treatment(P<0.05),but there was no difference between the two groups after treatment(P>0.05).In terms of mental health,there was no difference before and after treatment in treatment plan 1(P>0.05),while there was difference before and after treatment in treatment plan 2(P<0.05),and there was no difference between groups after treatment(P>0.05).(6)Inflammatory factors:①IL-1β:there was no difference between groups A and B before and after treatment(P>0.05),and there was no difference between groups after treatment(P>0.05).②IL-6 and IL-10:there were differences in group A before and after treatment(P<0.05),there were no differences in group B before and after treatment(P>0.05),and there were differences between groups after treatment(P<0.05).③TNF-α:There were differences between groups A and B before and after treatment(P<0.05),and there were also differences between groups after treatment(P<0.05).There was no significant difference in changes in blood and urine routine,liver and kidney function,and no significant adverse reactions were observed between the two groups of patients.Study 2.Chemical components identification of Qishentaohong granules.Methods:Ultrahigh performance liquid chromatography Q active focus mass spectrometry(UHPLC-QE-MS)was used to detect the chemical components of Qishen Taohong granules.In addition,the main components were selected based on the similarity score of the spectrogram,oral bioavailability(OB)>30%,and drug like properties(DL)>0.18.Combined with literature reports,the main components and its pharmacological effects of Qishentaohong granules were analyzed.Results:This study screened 13 main components of Qishen Taohong granules:astragaloside Ⅳ,quercetin,kaempferol,isorhamnetin,niacin,ginsenoside Rg3,ginsenoside F1,tanshinone ⅡA,cryptotanshinone,tanshinone,isoimperatorin,protocatechualdehyde,and baicalin.These components treat cardiovascular diseases through various mechanisms,particularly in immune regulation and anti-inflammatory.Study 3.Study on the immune regulation mechanism of Qishentaohong granules on rats with heart failure.Methods:A rat model of heart failure after myocardial infarction was established by ligating the left anterior descending branch of the coronary artery.The model was randomly divided into:①sham group(Sham,equal amount of distilled water),② model group(Model,equal amount of distilled water),③low dose group of Qishentaohong granules(QTG-L,Qishentaohong granules 16.8g/kg/d),④high dose group of Qishentaohong granules(QTG-H,33.6g/kg/d),⑤ fosinopril group(Fosinopril,4.2mg/kg/d),with 12 rats in each group.The intervention period was 4 weeks.Echocardiography was used to detect cardiac structure and function.The pathological changes of heart and spleen tissues were observed using hematoxylin/eosin and Masson staining.T cell subpopulation level was detected by flow cytometry.The α-SMA expression was detected by immunohistochemistry.Immunofluorescence staining was used to evaluate M1 and M2 macrophages.Enzyme-linked immunosorbent assay(ELISA)was used to measure the levels of Ang Ⅱ,NT-proBNP,IL-1β,IL-6,IL-10 and TNF-α.RTqPCR and Western blot were used to assess the mRNA and protein expression of AT1/MCP-1 in spleen tissue,as well as AT1/MCP-1/CCR2 and TGF-β/Smad3,VEGF,and CD31 in myocardial tissues.Results:1.Pathological staining of myocardial tissue:compared with the Sham group,the number of myocardial cells in the Model group was decreased,myocardial cells were swelled,inflammatory cells were infiltrated,and myocardial cells were filled with many myocardial collagen fibers.Compared with the Model group,these pathological changes of myocardial tissues in the QTG-L,QTG-H and Fosinopril groups were improved.2.Cardiac structure and function:compared with the Sham group,the levels of LVEF,LVFS,LVAWd,LVAWs,LVPWd and LVPWs in the Model group were decreased(P<0.05).Compared with the Model group,the levels of LVEF,LVFS,LVPWd and LVPWs in the QTG-L,QTG-H and Fosinopril groups were increased(P<0.05),while the levels of LVAWd and LVAWs in the QTG-H group were increased(P<0.05).3.NT-proBNP and Ang Ⅱ:compared with the Sham group,the levels of NT-proBNP and Ang Ⅱ in the Model group were increased(P<0.05).Compared with the Model group,the levels of NT-proBNP and Ang Ⅱ in the QTG-L,QTG-H and Fosinopril groups were decreased(P<0.05).4.Peripheral blood T lymphocyte subsets:compared with the Sham group,the levels of CD3+ and CD3+CD4+in the Model group were decreased(P<0.05).After QTG-L,QTG-H and Fosinopril intervention,the levels of CD3+and CD3+CD4+were increased(P<0.05)in each group.Compared with the Sham group,the Model group showed an increase in CD3+CD8+(P<0.05).After QTG-L and QTG-H intervention,the level of CD3+CD8+in each group decreased(P<0.05).5.Monocyte spleen mobilization:(1)spleen H&E staining:compared with the Sham group,the subcapsular red pulp of spleen tissue in the Model group was significantly thinner,and the number of cells was reduced,which showed many blank areas left after monocyte mobilization.Compared with the Model group,these pathological changes of spleen tissues in the QTG-L,QTG-H and Fosinopril groups were all improved.(2)AT1/MCP-1 in spleen tissue:compared with the Sham group,the levels of AT1,MCP-1 mRNA and protein in the Model group were increased(P<0.05).Compared with the model group,the levels of AT1 and MCP-1 mRNA and protein in QTG-L,QTG-H and Fosinopril groups were decreased(P<0.05).6.M1 macrophages in myocardial tissues:(1)M1 macrophages:compared with the Sham group,M1 macrophages in the Model group were increased.After the intervention of QTG-L,QTG-H and Fosinopril,the number of M1 macrophages in each group was decreased.(2)IL-1β,IL-6 and TNF-α:compared with the Sham group,the levels of IL-1β,IL-6 and TNF-α in the Model group were increased(P<0.05).Compared with the Model group,the levels of IL-1β and IL-6 in the QTG-L,QTG-H,Fosinopril groups were decreased(P<0.05),only TNF-α in the QTG-H group was decrease(P<0.05).(3)Myocardial tissue α-SAM and MMP2:compared with the Sham group,the levels of α-SAM and MMP2 in the Model group were increased(P<0.05).Compared with the Model group,the levels of α-SAM and MMP2 in the QTG-L,QTG-H and Fosinopril groups were decreased(P<0.05).(4)Cardiac tissue AT1/MCP-1/CCR2,TGF-β/Smad3 in myocardial tissues:compared with the Sham group,the levels of AT1,MCP-1,CCR2,TGF-β mRNA and protein in the Model group were increased(P<0.05).Compared with the Model group,the levels of AT1,MCP-1,CCR2,TGF-βmRNA and protein,Smad3 mRNA and p-Smad3 protein in the QTG-L,QTG-H and Fosinopril groups were decreased(P<0.05).7.M2 macrophages in myocardial tissue:(1)M2 macrophages:compared with the Sham group,M2 macrophages in the Model group were decreased.After the intervention of QTG-L,QTG-H and Fosinopril,M2 macrophages were increased in each group.(2)Serum IL-10:compared with the Sham group,the level of IL-10 in the Model group was decreased(P<0.05).Compared with the model group,the level of IL-10 in the QTG-L,QTG-H and Fosinopril groups were increased(P<0.05).(3)VEGF and CD31 in myocardial tissue:compared with the Sham group,the levels of VEGF and CD31 mRNA and protein in the Model group were decreased(P<0.05).Compared with the model group,the mRNA and protein of CD31 and VEGF in thr QTG-L,QTG-H and Fosinopril groups were increased(P<0.05).Conclusion:1.Qishentaohong granules has good efficacy and safety in the treatment of chronic HF and reduced inflammatory reaction.2.The main components of Qishentaohong granules have immunomodulatory and anti-inflammatory pharmacological effects.3.Qishentaohong granules improved cardiac function and myocardial remodeling by inhibiting the over-activation of Ang Ⅱ and regulating the immune and inflammatory reactions mediated by monocytes/macrophages and T cells. |
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