Objective To investigate the effects and mechanism of Linggui Qihua Prescription against myocardial inflammation and fibrosis in heart failure with preserved ejection fraction(HFpEF).Methods Totally 40 spontaneously hypertensive rats(SHR)were equally divided into model group,Entresto group(18 mg/kg)and Linggui Qihua Prescription low-and high-dosage groups(3.87,7.74 g/kg),the HFpEF model was induced using a 16-week high fat-salt-sugar diet and 8-week streptozotocin intraperitoneal injection.Ten WKY rats(WKY group)and 10 SHR(SHR group)served as control.After successful modeling,the groups were subjected to a 6-week corresponding intervention.Left ventricular end diastolic diameter(LVEDD),left ventricular end-diastolic volume(LVEDV),relative ventricular wall thickness(RWT),left ventricular fractional shortening(LVFS),isovolumic relaxation time(IVRT),mitral annular early and late diastolic velocities of motion(e? and a?),global longitudinal strain(GLS)and global longitudinal strain rate(GLSr)were measured by echocardiography;serum tumor necrosis factor-α(TNF-α)and soluble growth stimulation expressed gene 2 protein(sST2)contents were detected by ELISA;the morphology of myocardial tissue was observed by HE staining;myocardial fibrosis was assessed by Masson staining;the mRNA and protein expressions of myocardial tissue vascular cell adhesion molecule-1(VCAM-1),intercellular adhesion molecule-1(ICAM-1),growth differentiation factor-15(GDF-15)and sST2 were detected by qPCR and Western blot.Results Compared with the WKY group and SHR group,the model group exhibited significant increase in LVEDV,RWT,IVRT,along with significant decrease in e?,a? and absolute values of GLS and GLSr(P<0.05,P<0.01),the serum contents of TNF-α and sST2 significantly increased(P<0.01);there was pronounced myocardial inflammatory infiltration and collagen fiber deposition,while the mRNA and protein expression of VCAM-1,ICAM-1,GDF-15 and sST2 significantly increased(P<0.05,P<0.01).Compared with the model group,Entresto group and Linggui Qihua Prescription low-and high-dosage groups demonstrated significant decrease in LVEDV,RWT,IVRT,along with significant increases in e?,a? and absolute values of GLS and GLSr(P<0.05,P<0.01),the serum contents of TNF-α and sST2 significantly decreased(P<0.01);myocardial inflammatory infiltration and collagen fiber deposition were reduced,the mRNA and protein expressions of ICAM-1,GDF-15 and sST2 decreased in Linggui Qihua Prescription high-dosage group(P<0.01).Conclusion Linggui Qihua Prescription may inhibit chronic inflammation and fibrosis of the myocardium in HFpEF rats by regulating the expressions of ICAM-1,VCAM-1,GDF-15 and sST2,improving cardiac remodeling and functional impairment.

Objective To investigate the effects and mechanism of Linggui Qihua Prescription against myocardial inflammation and fibrosis in heart failure with preserved ejection fraction(HFpEF).Methods Totally 40 spontaneously hypertensive rats(SHR)were equally divided into model group,Entresto group(18 mg/kg)and Linggui Qihua Prescription low-and high-dosage groups(3.87,7.74 g/kg),the HFpEF model was induced using a 16-week high fat-salt-sugar diet and 8-week streptozotocin intraperitoneal injection.Ten WKY rats(WKY group)and 10 SHR(SHR group)served as control.After successful modeling,the groups were subjected to a 6-week corresponding intervention.Left ventricular end diastolic diameter(LVEDD),left ventricular end-diastolic volume(LVEDV),relative ventricular wall thickness(RWT),left ventricular fractional shortening(LVFS),isovolumic relaxation time(IVRT),mitral annular early and late diastolic velocities of motion(e? and a?),global longitudinal strain(GLS)and global longitudinal strain rate(GLSr)were measured by echocardiography;serum tumor necrosis factor-α(TNF-α)and soluble growth stimulation expressed gene 2 protein(sST2)contents were detected by ELISA;the morphology of myocardial tissue was observed by HE staining;myocardial fibrosis was assessed by Masson staining;the mRNA and protein expressions of myocardial tissue vascular cell adhesion molecule-1(VCAM-1),intercellular adhesion molecule-1(ICAM-1),growth differentiation factor-15(GDF-15)and sST2 were detected by qPCR and Western blot.Results Compared with the WKY group and SHR group,the model group exhibited significant increase in LVEDV,RWT,IVRT,along with significant decrease in e?,a? and absolute values of GLS and GLSr(P<0.05,P<0.01),the serum contents of TNF-α and sST2 significantly increased(P<0.01);there was pronounced myocardial inflammatory infiltration and collagen fiber deposition,while the mRNA and protein expression of VCAM-1,ICAM-1,GDF-15 and sST2 significantly increased(P<0.05,P<0.01).Compared with the model group,Entresto group and Linggui Qihua Prescription low-and high-dosage groups demonstrated significant decrease in LVEDV,RWT,IVRT,along with significant increases in e?,a? and absolute values of GLS and GLSr(P<0.05,P<0.01),the serum contents of TNF-α and sST2 significantly decreased(P<0.01);myocardial inflammatory infiltration and collagen fiber deposition were reduced,the mRNA and protein expressions of ICAM-1,GDF-15 and sST2 decreased in Linggui Qihua Prescription high-dosage group(P<0.01).Conclusion Linggui Qihua Prescription may inhibit chronic inflammation and fibrosis of the myocardium in HFpEF rats by regulating the expressions of ICAM-1,VCAM-1,GDF-15 and sST2,improving cardiac remodeling and functional impairment.

Objective To investigate the efficacy and safety of Linggui Qihua No.2 Prescription(LGQH2)in patients with heart failure with preserved ejection fraction(HFpEF).Methods Totally 60 HFpEF patients were randomly divided into experimental group and control group according to random number table method,with 30 patients in each group.On the basis of standardized treatment for heart failure,the experimental group was given LGQH2 granules,13 g/time,twice a day,orally;the control group was given placebo granules of LGQH2,with the same administration method as the experimental group.The treatment course for both groups was 4 weeks.6-minute walking distance(6MWD),Kansas City Cardiomyopathy Questionnaire(KCCQ)score,TCM syndrome score and serum NT-proBNP levels.Echocardiography was used to detect the ratio of early diastolic blood flow velocity(E)at the mitral valve to early diastolic myocardial motion velocity(e')at the mitral annulus,left atrial diameter(LAD),left ventricular end-diastolic diameter(LVEDD)and interventricular septal thickness(IVST).Adverse reactions and events were also recorded.Results Compared with before treatment,both groups showed significant improvement in 6MWD,KCCQ score,TCM syndrome score,serum NT proBNP,and E/e'after treatment(P<0.05),and there was no statistical significance in LAD,LVEDD and IVST between the two groups(P>0.05);after treatment,the experimental group showed better improvement in 6MWD,KCCQ score,TCM syndrome score and E/e'compared to the control group(P<0.05).During the research process,neither group of patients experienced any adverse reactions or events.Conclusion LGQH2 Prescription can effectively enhance exercise tolerance and cardiac function of HFpEF patients,alleviate symptoms,improve quality of life,and inhibit diastolic dysfunction of the heart,without notable adverse reactions.

Objective To develop a core outcome set(COS)for clinical effectiveness trials of heart failure with preserved ejection fraction(HFpEF).Methods Outcome measures were collected through database literatures search,clinical experts questionnaire survey and semi-structured patients interview.Then,the outcome measures pool was constructed and domains were divided.Candidate outcome measures of COS were screened through two rounds of Delphi survey.Finally,a consensus meeting was held to determine COS and reach a consensus.Results A total of 317 outcome measures which could be divided into 6 domains were collected through literature research,questionnaire survey and semi-structured interview.15 candidate outcome measures of COS were screened through two rounds of Delphi survey.Finally,the consensus meeting reached consensus on a COS with 6 entries.Conclusion In this study,a COS for clinical effectiveness trials of HFpEF was developed,which is conducive to the standardization of efficacy evaluation.

Objective To develop a core outcome set(COS)for clinical effectiveness trials of heart failure with preserved ejection fraction(HFpEF).Methods Outcome measures were collected through database literatures search,clinical experts questionnaire survey and semi-structured patients interview.Then,the outcome measures pool was constructed and domains were divided.Candidate outcome measures of COS were screened through two rounds of Delphi survey.Finally,a consensus meeting was held to determine COS and reach a consensus.Results A total of 317 outcome measures which could be divided into 6 domains were collected through literature research,questionnaire survey and semi-structured interview.15 candidate outcome measures of COS were screened through two rounds of Delphi survey.Finally,the consensus meeting reached consensus on a COS with 6 entries.Conclusion In this study,a COS for clinical effectiveness trials of HFpEF was developed,which is conducive to the standardization of efficacy evaluation.

Objective To investigate the efficacy and safety of Linggui Qihua No.2 Prescription(LGQH2)in patients with heart failure with preserved ejection fraction(HFpEF).Methods Totally 60 HFpEF patients were randomly divided into experimental group and control group according to random number table method,with 30 patients in each group.On the basis of standardized treatment for heart failure,the experimental group was given LGQH2 granules,13 g/time,twice a day,orally;the control group was given placebo granules of LGQH2,with the same administration method as the experimental group.The treatment course for both groups was 4 weeks.6-minute walking distance(6MWD),Kansas City Cardiomyopathy Questionnaire(KCCQ)score,TCM syndrome score and serum NT-proBNP levels.Echocardiography was used to detect the ratio of early diastolic blood flow velocity(E)at the mitral valve to early diastolic myocardial motion velocity(e')at the mitral annulus,left atrial diameter(LAD),left ventricular end-diastolic diameter(LVEDD)and interventricular septal thickness(IVST).Adverse reactions and events were also recorded.Results Compared with before treatment,both groups showed significant improvement in 6MWD,KCCQ score,TCM syndrome score,serum NT proBNP,and E/e'after treatment(P<0.05),and there was no statistical significance in LAD,LVEDD and IVST between the two groups(P>0.05);after treatment,the experimental group showed better improvement in 6MWD,KCCQ score,TCM syndrome score and E/e'compared to the control group(P<0.05).During the research process,neither group of patients experienced any adverse reactions or events.Conclusion LGQH2 Prescription can effectively enhance exercise tolerance and cardiac function of HFpEF patients,alleviate symptoms,improve quality of life,and inhibit diastolic dysfunction of the heart,without notable adverse reactions.

Objective To evaluate the characteristics of a rat model of heart failure with a preserved ejection fraction(HFpEF)induced by combined factors,and to investigate the correlation of myocardial strain parameters to myocardial hypertrophy and fibrosis.Methods Eight WKY rats and eight spontaneously hypertensive rats(SHR)served as control groups and were fed normal feed until the end of the experiment.Thirty-two SHR rats were equally divided into SHR+S,SHR+F,SHR+SF,and SHR+Combined groups,and fed high-salt,high-fat,high-salt-fat,or high-salt-fat-sugar feed,respectively,in combination with intraperitoneal injection of streptozotocin for 30 weeks.After modeling,the heart weight/body weight(HW/BW)ratio,systolic blood pressure(SBP),and diastolic blood pressure(DBP)were measured.Echocardiography was performed to measure the left ventricular(LV)end-diastolic internal diameter(LVIDd),LV anterior wall thickness(LVAWd),LV posterior wall thickness(LVPWd),LV ejection fraction(LVEF),isovolumetric diastolic time(IVRT),and peak early diastolic passive filling velocity(E)/early diastolic mitral annular velocity(e').Speckle tracking echocardiography was conducted to determine the global longitudinal strain(GLS)and strain rate(GLSr),global radial strain(GRS)and strain rate(GRSr),as well as the global circumferential strain(GCS)and strain rate(GCSr).Serum was collected and analyzed for triglycerides(TG),total cholesterol(TC),low-density lipoprotein cholesterol(LDL-C),glucose(GLU),and glycated serum protein(GSP).ELISA were used to measure serum B-type brain natriuretic peptide(BNP),angiotensin Ⅱ(AngⅡ),and galectin-3(Gal-3).Myocardial tissue was subjected to HE and Masson staining for cardiomyocytes and myocardial fibrosis,and the cardiomyocyte cross-sectional area(CSA)and collagen volume fraction(CVF)were calculated.Additionally,the correlation of myocardial strain parameters to CSA and CVF was analyzed.Results Compared with the control group,in model groups,especially the SHR+combined group,HW/BW,SBP,DBP,serum indexes(TC,TG,LDL-C,GLU,GSP,BNP,AngⅡ,and Gal-3)and echocardiographic parameters(LVIDd,LVAWd,LVPWd,IVRT,and E/e')were significantly up-regulated.Absolute values of speckle-tracking echocardiographic parameters(GLS,GLSr,GRS,GRSr,GCS,and GCSr)were decreased considerably.HE and Masson staining of myocardial tissues suggested marked cardiomyocyte hypertrophy and fibrosis,and significant increases were observed in CSA and CVF(P＜0.05).Correlation analysis showed that GLSr,GCS,and GCSr were strongly linked to CSA,and GLS,GLSr,and GCSr were strongly linked to CVF(P＜0.01).Conclusions A rat model of HFpEF induced by hypertension and dysregulation of glucolipid metabolism replicated the basic characteristics of HFpEF in terms of etiology,clinical features,and myocardial pathological changes,and might be a reliable animal model of metabolic syndrome-related HFpEF.Moreover,myocardial strain indices were closely related to myocardial hypertrophy and fibrosis and might indirectly reflect subtle myocardial lesions and dysfunction.

Objective:To investigate whether Lycopi Herba can serve as a viable alternative to Alismatis Rhizoma in the treatment of heart failure (HF) through network pharmacology and molecular docking techniques.Methods:TCMSP database was used to filter active components of Lycopi Herba and Alismatis Rhizoma. SwissTargetPrediction database was used to predict potential targets. HF-related targets were collected from databases such as GeneCards, OMIM, and DisGeNET. Venny 2.1.0 was used to draw a Venn diagram illustrating the intersection of targets between Lycopi Herba and Alismatis Rhizoma and HF. A protein-protein interaction (PPI) network was established using the String database, and key targets for the treatment of HF with Lycopi Herba and Alismatis Rhizoma were selected using Cytoscape 3.9.1 software to construct a component-intersection target network. The intersection targets were then analyzed for Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways using Metascape. Molecular docking techniques were used to evaluate the affinity between active components and key targets.Results:Lycopi Herba primarily targeted pivotal proteins such as HMGCR and CYP27B1, while Alismatis Rhizoma had a broader target spectrum, including PPARA, JAK2, among others. Shared key targets between the two included HMGCR and ESR1, which were primarily involved in cholesterol synthesis and steroid hormone biosynthesis. Enrichment pathway analysis showed similarities in steroid metabolism between the two; Alismatis Rhizoma, however, was more likely to act through protein phosphorylation regulation and modulating the PI3K-Akt signaling pathway for HF treatment. A unique target for Lycopi Herba in treating HF was CHRM4, indicating its potential for blood pressure regulation and myocardial protection.Conclusions:Both Lycopi Herba and Alismatis Rhizoma exhibit certain commonalities in the treatment of HF, but Alismatis Rhizoma has a wider range of targets and signaling pathways, implying more extensive therapeutic potential. However, considering the nephrotoxicity of Alismatis Rhizoma, Lycopi Herba could be considered as an alternative treatment for HF, especially in patients with renal insufficiency or in the early stages of HF.

ObjectiveTo explore the material basis and molecular mechanism of Linggui Qihua prescription （LGQH） against myocardial fibrosis in heart failure with preserved ejection fraction （HFpEF）. MethodLiquid chromatography-mass spectrometry （LC-MS） was used to qualitatively analyze the active components of LGQH. AutoDock software was employed for molecular docking between the active components of LGQH and target proteins including α-smooth muscle actin （α-SMA）， type Ⅰ collagen （ColⅠ）， type Ⅲ collagen （ColⅢ）， matrix metalloproteinase-9 （MMP-9）， and tissue inhibitor of metalloproteinase-1 （TIMP-1）. In vivo experiments were conducted on 40 spontaneously hypertensive rats （SHRs） aged 4 weeks， which were divided into an HFpEF group， an Entresto group （0.018 g·kg^-1）， and low- and high-dose LGQH groups （3.87， 7.74 g·kg^-1）. A high-fat， high-salt， and high-sugar diet was administered for 16 weeks along with intraperitoneal injection of streptozotocin solution for 8 weeks to establish an HFpEF model in rats. The blank group consisted of 10 Wistar Kyoto （WKY） rats and 10 SHRs. After successful modeling， the WKY， SHR， and HFpEF groups were given equal volumes of normal saline， while the other three groups received predetermined interventions. Daily oral gavage was performed for 6 weeks. After intervention， echocardiography was conducted to measure left ventricular （LV） anterior wall thickness （LVAWd）， LV posterior wall thickness （LVPWd）， LV internal diameter at end-diastole （LVIDd）， LV ejection fraction （LVEF）， isovolumic relaxation time （IVRT）， early diastolic peak velocity of mitral valve inflow （E）， and early diastolic mitral annular velocity （e'）. The E/e' ratio was calculated. Enzyme-linked immunosorbent assay （ELISA） was used to detect serum atrial natriuretic peptide （ANP）， B-type natriuretic peptide （BNP）， and galectin-3 （Gal-3）. Myocardial fibrosis was observed through Masson staining of pathological sections， and collagen volume fraction （CVF） and perivascular fibrosis ratio （PFR） were calculated. Real-time polymerase chain reaction （PCR） and Western blot were employed to detect LV myocardial mRNA and protein expression of α-SMA， ColⅠ， ColⅢ， MMP-9， and TIMP-1. ResultLC-MS identified 13 active components in LGQH. Molecular docking indicated stable binding of the 13 compounds with five target proteins. In vivo experiments showed that compared with the blank group， the HFpEF group had significantly increased LVAWd， LVPWd， LVIDd， IVRT， E/e'， ANP， BNP， Gal-3， CVF， and PFR. LV myocardial α-SMA， ColⅠ， and ColⅢ mRNA and protein expression was significantly upregulated， while MMP-9/TIMP-1 mRNA and protein ratios were significantly downregulated （P<0.05， P<0.01）. Compared with the HFpEF group， LGQH might dose-dependently reduce LVAWd， LVPWd， LVIDd， IVRT， E/e'， ANP， BNP， Gal-3， CVF， and PFR， downregulated myocardial α-SMA， ColⅠ， ColⅢ mRNA expression， α-SMA， and ColⅠ protein expression， and upregulated MMP-9/TIMP-1 mRNA and protein expression （P<0.05， P<0.01）. ConclusionLGQH contains multiple active components and may inhibit myocardial fibrosis in HFpEF rats. It may further alleviate LV hypertrophy， dilation， and diastolic dysfunction， making it an effective Chinese medicinal prescription for treating HFpEF.

Objective:To evaluate the pathological features of a heart failure with preserved ejection fraction(HFpEF) model, which is established by spontaneously hypertensive rats (SHR) through high-fat diet and diabetic factors.Methods:Twenty specific pathogen-free grade(SPF grade) and 14-week-old SHR rats were randomly divided into SHR group (normal diet) and HFpEF group [high-fat diet combined with intraperitoneal injection of streptozotocin (STZ, 25 mg/kg) were used to create a diabetic complex model] with 10 rats in each group. Ten SPF and 14-week-old WKY rats with the same genetic background were set as blank control group (WKY group). All rats were fed for 8 weeks. Echocardiography was performed to measure cardiac parameters: peak velocity of early diastolic mitral inflow(E), peak velocity of late diastolic mitral inflow(A), and the early diastolic mitral annulus e′ in the same cardiac cycle, left atrial ejection fraction (LAEF), left ventricular ejection fraction (LVEF), left atrial diameter, right atrial diameter and interventricular septal thickness(IVST). Serological testing included glucose (GLU) and glycosylated serum protein (GSP); Enzyme-linked immunoassay (ELISA) testing included insulin (INS), glucagon (PG), C-peptide (CP), leptin (LEP), atrial natriuretic peptide (ANP) and B-type brain natriuretic peptide (BNP). The rat heart tissue was stained with HE, and the morphological changes of atrial/ventricular tissue were observed under an optical microscope.Results:The pathological characteristics of HFpEF was established in SHR rats fed with high fat and diabetes. Echocardiography showed that compared with the WKY group, the values of E, E/A and E/e′ in the HFpEF group were significantly increased (all P<0.01), and e′and LAEF were significantly reduced (all P<0.01). In the HFpEF group, the anteroposterior and tranverse dimensions of the left atrium and the long-axis dimension of the right atrium increased to varying degrees (all P<0.05), and the IVST was also significantly increased ( P<0.01). At the same time, atrial wall was thickened obviously, myocardial cells were disordered, and myocardial fibers were broken. Compared with the WKY group, the levels of serum markers ANP and BNP in HFpEF group were significantly increased (all P<0.01), and the levels of serum insulin-related indicators INS, PG, CP, LEP, GSP, and GLU increased to varying degrees (all P<0.01). Conclusions:The composite model established by SHR rats through high-fat diet and diabetic factors can simulate the Doppler echocardiographic changes and pathological features of HFpEF, as well as abnormal changes in serum related markers and insulin indicators.