Application of tilianin in preparation of medicine for preventing and / or treating ejection fraction retention type heart failure
By regulating the PI3K/AKT signaling pathway with piracetamidine, drugs of different dosage forms were prepared, which solved the problems of left ventricular diastolic dysfunction, myocardial damage and liver damage in heart failure with preserved ejection fraction, and achieved significant therapeutic effects.
Patent Information
- Application Number
- CN202511921981.8
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2025-12-17
- Publication Date
- 2026-02-10
AI Technical Summary
There is a lack of effective drugs for treating heart failure with preserved ejection fraction, especially for myocardial damage, cardiac hypertrophy, and liver damage caused by heart failure with preserved ejection fraction induced by N-nitro-L-arginine methyl ester and a high-fat diet.
Using tilianin as the active ingredient, drugs in different dosage forms, including injections, capsules, and tablets, are prepared to regulate the PI3K/AKT signaling pathway, improve left ventricular diastolic dysfunction in heart failure with preserved ejection fraction, inhibit myocardial fibrosis and cardiomyocyte hypertrophy, and improve hepatic steatosis and blood lipid levels.
This herbal extract significantly reduced mean arterial pressure in mice, improved left ventricular diastolic function, reduced myocardial fibrosis area, decreased inflammatory cell infiltration, improved cardiomyocyte structure, reduced serum lipid levels and hepatic steatosis, providing multidimensional therapeutic effects.
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Figure CN121489968A_ABST
Abstract
Description
Technical Field
[0001] This invention belongs to the field of biopharmaceutical technology, specifically relating to the application of piracetam in the preparation of drugs for the prevention and / or treatment of heart failure with preserved ejection fraction. Background Technology
[0002] Heart failure is a complex clinical syndrome caused by various factors leading to abnormal changes in the structure and / or function of the heart, resulting in impaired ventricular systolic and / or diastolic function. Based on differences in left ventricular ejection fraction (LVEF) and changes after treatment, heart failure is classified into four categories: heart failure with reduced ejection fraction (HFrEF), heart failure with improved ejection fraction (HFimpEF), heart failure with mildly reduced ejection fraction (HFmrEF), and heart failure with preserved ejection fraction (HFpEF). Epidemiological studies indicate that more than 50% of heart failure patients have HFpEF. HFpEF is associated with hypertension, diabetes, lipid metabolism disorders, obesity, and aging. However, there are currently no effective drugs for treating HFpEF.
[0003] Tilianin is one of the main active monomers in *Xiangqinglan*, a traditional Chinese medicine from Xinjiang. It belongs to the flavonoid class of compounds. Current research suggests that tilianin has a certain effect in treating ischemic perfusion re-injury and arteriosclerosis. However, its therapeutic effect on heart failure with preserved ejection fraction has not been reported. Summary of the Invention
[0004] The purpose of this invention is to provide a new pharmaceutical use for citronellol, specifically its use in the preparation of drugs for the prevention and / or treatment of heart failure with preserved ejection fraction.
[0005] This invention provides the use of piracetam in the preparation of medicaments for the prevention and / or treatment of heart failure with preserved ejection fraction.
[0006] Preferably, the ejection fraction-preserving heart failure includes ejection fraction-preserving heart failure induced by N-nitro-L-arginine methyl ester (L-NAME) and a high-fat diet.
[0007] This invention provides the use of piracetam in the preparation of medicaments for the prevention and / or treatment of left ventricular diastolic dysfunction caused by heart failure with preserved ejection fraction.
[0008] This invention provides the use of cirsinoside in the preparation of drugs for the prevention and / or treatment of myocardial damage or cardiac hypertrophy as complications of heart failure with preserved ejection fraction.
[0009] Preferably, the myocardial injury complication of the ejection fraction-preserving heart failure includes at least one of the following conditions: cardiomyocyte disorder, myocardial hypertrophy, and myocardial fibrosis.
[0010] This invention provides the use of cirsinoside in the preparation of medicaments for the prevention and / or treatment of liver damage as a complication of heart failure with preserved ejection fraction.
[0011] Preferably, the liver damage associated with the preservative ejection fraction heart failure includes hepatic steatosis and / or elevated blood lipid levels.
[0012] Preferably, the drug comprises at least one of the following dosage forms: injection, capsule, tablet, powder, granule, pill, microcapsule / microsphere formulation, suppository, ointment, spray, and targeted formulation.
[0013] Preferably, the drug further includes pharmaceutically acceptable excipients; The mass percentage of citronellol in the drug is 0.1% to 99.5%.
[0014] This invention provides the application of argiol in the preparation of drugs for the prevention and / or treatment of heart failure with preserved ejection fraction. Using a mouse model of heart failure with preserved ejection fraction induced by L-NAME and a high-fat diet, this invention conducted a pharmacodynamic experiment on the prevention and treatment of heart failure with preserved ejection fraction using argiol. The results showed that the systolic blood pressure, diastolic blood pressure, and mean arterial pressure of the treated mice were significantly lower than those of the model group; the E / E′ ratio of the treated mice was significantly lower than that of the model group; and the thickness of the interventricular septum at end-diastole and end-systole, the thickness of the left ventricular posterior wall at end-diastole and end-systole, and the mass of the left ventricle were significantly different between the treated mice and the model group, indicating that argiol can improve diastolic dysfunction of the left ventricle in the mouse model. Simultaneously, H&E staining results showed that the cardiomyocytes in the treatment group mice were disordered and inflammatory cells were infiltrated, while the tissue structure of the treatment group mice tended to be normal. Masson staining results showed that the area of myocardial fibrosis in the model group mice was significantly increased, while the area of fibrosis in the treatment group mice was significantly reduced. WGA staining results indicated that the cross-sectional area of cardiomyocytes in the model group was increased, while cell hypertrophy in the treatment group mice was significantly improved. This indicates that the described arilin has a good inhibitory effect on myocardial injury or cardiac hypertrophy, has a cardioprotective effect, and also has a certain ameliorative effect on liver damage, a complication of heart failure with preserved ejection fraction. In addition, WB results showed that arilin exerts its therapeutic effect by regulating the PI3K / AKT signaling pathway. Therefore, this invention provides a new approach for the treatment of heart failure with preserved ejection fraction and also broadens the pharmaceutical applications of arilin. Attached Figure Description
[0015] Figure 1The results of this study on the improvement of routine parameters in HFpEF mice by piracetam are shown below. A: Animal experimental dosing regimen; B: Comparison of body weight between different mouse groups; C, D, and E: Changes in systolic blood pressure, diastolic blood pressure, and mean arterial pressure in different experimental groups, respectively; F and G: Blood glucose concentration change curves and area under the curve (AUC) of the glucose tolerance test, respectively; H and I: Heart weight and the ratio of heart weight to tibia length, respectively; J and K: Lung weight and the ratio of lung weight to tibia length, respectively; "HFpEF" represents the model control group mice, and "Til" represents the model drug-treated group mice; ns indicates no significant difference between the two groups. P >0.05; This indicates a significant difference between the two groups. P <0.05; This indicates a significant difference between the two groups. P <0.01; This indicates a significant difference between the two groups. P <0.005; Figure 2 Results of Til administration in improving left ventricular diastolic dysfunction in HFpEF mice; A: Representative images of mice: routine echocardiography and Doppler imaging; B: Left ventricular ejection fraction; C: Left ventricular shortening fraction; D: Ratio of mitral valve E wave to E′ wave; E: End-diastolic interventricular septal wall thickness; F: End-systolic interventricular septal wall thickness; G: Left ventricular posterior wall at end-diastole; H: Left ventricular posterior wall at end-systole; I: Left ventricular mass; "Control" represents normal mice, "HFpEF" represents model control mice, and "Til" represents model administration mice; ns indicates no statistical significance between the two groups. P >0.05; This indicates a significant difference between the two groups. P <0.05; This indicates a significant difference between the two groups. P <0.01; This indicates a significant difference between the two groups. P <0.005; Figure 3The results show the effects of Til on inhibiting myocardial disarray, hypertrophy, and fibrosis in HFpEF mice. Figure A shows representative images of H&E staining of left ventricular cardiomyocytes in different groups (scale bar = 20 µm), representative images of Masson staining of left ventricular cardiomyocytes in different groups (scale bar = 50 µm), and representative images of wheat germ lectin (WGA) staining of left ventricular cardiomyocytes in different groups (scale bar = 20 µm). Figure B: Statistical graph of cardiac collagen volume; Figure C: Statistical graph of cardiomyocyte cross-sectional area. "Control" represents normal mice, "HFpEF" represents model control mice, and "Til" represents model drug-treated mice. ns indicates no significant difference between the two groups. P >0.05; This indicates a significant difference between the two groups. P <0.05; This indicates a significant difference between the two groups. P <0.005; Figure 4 Image showing the improvement of hepatic steatosis and lipid droplet distribution in HFpEF mice after Til administration; A: Representative images of liver H&E staining in different groups (scale bar = 50µm), and representative images of liver Oil Red staining in different groups (scale bar = 50µm); B: Statistical graph of Oil Red staining area; C: Low-density lipoprotein; D: Serum total cholesterol; E: Serum triglycerides; F: Serum aspartate aminotransferase; G: Serum alanine aminotransferase; H: Liver total cholesterol; I: Liver triglycerides; "Control" represents normal mice, "HFpEF" represents model control mice, and "Til" represents model administration group mice; ns indicates no significant difference between the two groups. P >0.05; This indicates a significant difference between the two groups. P <0.05; This indicates a significant difference between the two groups. P <0.01; This indicates a significant difference between the two groups. P <0.005; Figure 5Diagram illustrating the potential mechanism by which Til administration improves HFpEF mice using transcriptomics; A: Cluster diagram of the top 20 differentially expressed genes between the normal and model groups in transcriptomics; B: Volcano diagram of differentially expressed genes between the model and administration groups in transcriptomics; C: BNP gene expression level; D: Ptprb gene expression level; E: CD19 gene expression level; F: Top 9 signaling pathways enriched by GSEA in transcriptomics; G: WB diagram of the PI3K / AKT signaling pathway; H: pPI3K / PI3K expression level; I: pAKT / AKT expression level; ns indicates no significant difference between the two groups. P >0.05; This indicates a significant difference between the two groups. P <0.05; This indicates a significant difference between the two groups. P <0.01; This indicates a significant difference between the two groups. P <0.005. Detailed Implementation
[0016] This invention provides the use of piracetam in the preparation of medicaments for the prevention and / or treatment of heart failure with preserved ejection fraction.
[0017] In this invention, the CAS number of the citronellol is 4291-60-5, the molecular weight is 446.404, and the molecular formula is C 22 H 22 O 10 The structural formula is shown in Formula I.
[0018] Formula I In this embodiment of the invention, the citronellol was purchased from Chengdu Lemeitian Pharmaceutical Technology Co., Ltd., with the product number DSTDT006702.
[0019] In this invention, the ejection fraction-preserving heart failure preferably includes ejection fraction-preserving heart failure induced by N-nitro-L-arginine methyl ester (L-NAME) and a high-fat diet. The preferred induction concentration of L-NAME is 0.5 g / L. The preferred high-fat diet is high-fat diet D12492. In one embodiment of this invention, a mouse model of ejection fraction-preserving heart failure was induced by N-nitro-L-arginine methyl ester (L-NAME) and a high-fat diet. Compared with mice in the normal diet group, the model group mice showed significantly increased body weight, significantly increased mean arterial pressure, significantly increased glucose tolerance, and significantly increased cardiac hypertrophy and lung mass. At the same time, there was no significant difference in ejection fraction and short-axis contraction rate between the normal group and the model group, but the E / E′ ratio was significantly higher than that of the model group. The interventricular septal thickness, left ventricular posterior wall thickness, and left ventricular mass were significantly increased at end-diastole and end-systole. Furthermore, myocardial tissue staining results showed that, compared with the normal control group, the model group exhibited disordered cardiomyocyte arrangement, inflammatory cell infiltration, and a significantly increased area of myocardial fibrosis, as well as an enlarged cross-sectional area of cardiomyocytes. Additionally, liver tissue staining revealed extensive vacuolation and hepatic steatosis in the liver tissue compared to the normal group. Serum levels of low-density lipoprotein, total cholesterol, triglycerides, aspartate aminotransferase, and alanine aminotransferase were significantly elevated in the model group compared to the normal group. In liver tissue, the expression levels of total cholesterol and triglycerides were significantly lower in the treatment group compared to the model group.
[0020] In this invention, the preferred concentration of cirsinoside is 10-20 mg / kg body weight in mice per day. Cirsinoside is administered via gavage. The gavage is preferably administered for 4 weeks.
[0021] In one embodiment of the present invention, the argentin significantly reduces the mean arterial pressure in model mice, improving cardiac hypertrophy, but has no effect on pulmonary edema. Furthermore, administration of the argentin reduces the E / E′ ratio in model mice, significantly reduces end-diastolic and end-systolic interventricular septal thickness, end-diastolic and end-systolic left ventricular posterior wall thickness, and left ventricular mass, effectively improving left ventricular diastolic dysfunction in model mice. Therefore, the present invention provides the application of argentin in the preparation of drugs for the prevention and / or treatment of left ventricular diastolic dysfunction caused by heart failure with preserved ejection fraction.
[0022] In this invention, the aralia elata glycoside can inhibit the occurrence of myocardial misalignment, myocardial hypertrophy, and myocardial fibrosis in HFpEF mice. In one embodiment of this invention, administration of the aralia elata glycoside improves the normalization of myocardial tissue structure, reduces inflammatory cell infiltration, and significantly reduces the area of myocardial fibrosis and improves myocardial cell hypertrophy. Therefore, this invention provides the use of aralia elata glycoside in the preparation of drugs for the prevention and / or treatment of myocardial injury or cardiac hypertrophy as complications of heart failure with preserved ejection fraction.
[0023] In this invention, liver injury in complications of heart failure with preserved ejection fraction (HFpEF) preferably includes hepatic steatosis and / or elevated blood lipid levels. Administration of the citronellol also improves hepatic steatosis and lipid droplet distribution in HFpEF mice. In one embodiment of this invention, compared to the model group, administration of the citronellol reduces hepatic vacuolation and lipid droplet area in mice, while also reducing serum low-density lipoprotein and total cholesterol levels, as well as alanine aminotransferase activity, and further reducing total cholesterol and triglyceride levels in liver tissue. Therefore, this invention provides the use of citronellol in the preparation of medicaments for the prevention and / or treatment of liver injury in complications of heart failure with preserved ejection fraction (HFpEF).
[0024] In this invention, the drug preferably comprises at least one of the following dosage forms: injection, capsule, tablet, powder, granule, pill, microcapsule / microsphere formulation, suppository, ointment, spray, and targeted formulation. The drug preferably also includes pharmaceutically acceptable excipients. The mass percentage of citronellol in the drug is preferably 0.1% to 99.5%, but can be 1% to 99%, 5% to 80%, 10% to 70%, 15% to 50%, further 20% to 40%, and preferably 30%. This invention does not impose any particular limitation on the preparation method of the drug; any drug preparation method well known in the art can be used.
[0025] The following examples illustrate the application of the cirsin provided by the present invention in the preparation of drugs for the prevention and / or treatment of heart failure with preserved ejection fraction, but these examples should not be construed as limiting the scope of protection of the present invention.
[0026] Example 1 Experiment on mouse model of heart failure with preserved ejection fraction (HFpEF) treated with citronellol 1. Materials and Reagents C57BL / 6N wild-type mice were purchased from Beijing Spaford, and the sources of the reagents are shown in Table 1 below.
[0027] Table 1. Source of Experimental Materials and Reagents
[0028] In addition to the above, all other materials and reagents used in this embodiment are commercially available products.
[0029] 2. Animal Experimentation Guidelines All animal studies were conducted under the guidance of the Experimental Animal Center of the Ethics Review Committee of Shihezi University. All mice were housed in the same environment, and were randomly grouped during the experiments. Echocardiographic analysis was performed by independent researchers unaware of the research objectives.
[0030] 3. Induction of a heart failure model with preserved ejection fraction C57BL / 6N mice were fed drinking water with a pH of 7.4 (containing 0.5 g / L L-NAME) for 10 consecutive weeks and induced with HFD (D12492 high-fat diet). Control mice were fed a standard diet and normal drinking water without L-NAME. After completing the 10-week diet regimen, cardiac function, blood pressure, and blood glucose were measured. When the ejection fraction (LVEF) remained unchanged, the model group showed an increase in E / E′ (the ratio of mitral valve orifice velocity to mitral valve annular velocity, used to assess left ventricular filling pressure and diastolic function) compared to the normal group, indicating diastolic dysfunction. Blood pressure showed a significant increase in diastolic pressure, systolic pressure, and mean arterial pressure compared to the normal group, and insulin resistance was observed, indicating successful model establishment.
[0031] Mice with similar baseline characteristics were then randomly assigned to four groups (n=8 per group): a control group (fed standard diet and given normal drinking water); an HFpEF model group (fed with HFD and L-NAME, administered via gavage for 4 weeks); a low-dose HFpEF + citronellol treatment group (fed with HFD and L-NAME, administered citronellol 10 mg / kg / day via gavage for 4 weeks); and a medium-dose HFpEF + citronellol treatment group (fed with HFD and L-NAME, administered citronellol 20 mg / kg / day via gavage for 4 weeks). Body weight and food intake were monitored weekly.
[0032] 4. The therapeutic effects of drugs on HFpEF The following measurements were taken after completing 4 weeks of treatment: 4.1 Routine Ultrasonic Testing Mice were anesthetized with oxygen and isoflurane (1%–2%). Left ventricular ejection fraction and other systolic function indices were obtained by short-axis M-mode scanning of the mid-ventricular segment. Diastolic function was measured at the mitral valve level using pulsed wave and tissue Doppler. Collected parameters included: end-diastolic interventricular septal wall thickness (IVSd), systolic interventricular septal wall thickness (IVSs), left ventricular end-diastolic posterior wall thickness (LVPWd), left ventricular end-systolic posterior wall thickness (LVPWs), left ventricular fractional shortening (LVFS), left ventricular ejection fraction (LVEF), peak Doppler velocity (E) of blood flow through the mitral valve in early diastole, peak tissue Doppler velocity (E') of myocardial diastolic velocity at the mitral valve annulus during early diastole, and left ventricular myocardial mass (LVmass).
[0033] 4.2 Blood Pressure Measurement Systolic blood pressure was measured noninvasively in conscious mice using a mouse blood pressure monitor (BP-2010 Series). Animals were placed in individual racks on a temperature-controlled platform (37°C) and data were recorded under steady-state conditions. All mice were trained to acclimatize to short-term restraint prior to testing. Blood pressure was recorded at least seven times for each mouse, and the readings were averaged.
[0034] 4.3 Staining 1. Tissue Acquisition and Fixation: After euthanizing C57 mice, the heart and liver were carefully removed. Fresh heart and liver tissue samples were placed in PBS and the blood was pumped out. The left ventricle of the heart tissue was separated, and the liquid was then blotted dry with absorbent paper (some samples were directly placed in a -80°C freezer for subsequent experiments). The tissue was then fixed in 4% paraformaldehyde. After 48 hours of fixation, the tissue was removed from the fixative.
[0035] 2. Dehydration, clearing, embedding, and sectioning: Heart tissue was sequentially dehydrated, cleared, embedded in paraffin, and sectioned (4-6 μm). Liver tissue was rapidly frozen in an OCT compound and sectioned using a cryostat (4-6 μm).
[0036] 3. Staining: HE staining: Heart tissue was stained using the conventional HE staining method, and myocardial morphological changes were observed under an optical microscope.
[0037] WGA staining: After dewaxing the heart tissue sections to water, WGA staining was performed (using fluorescently labeled WGA, and the sections were mounted after staining), and the distribution of myocardial cell membranes and glycoproteins was observed under a fluorescence microscope.
[0038] Masson staining: Heart tissue was stained using the conventional Masson trichrome staining method, and morphological changes such as collagen fibers were observed under an optical microscope.
[0039] Oil Red O staining: Liver tissue was stained using the conventional Oil Red O staining method, and changes in lipid droplet morphology were observed under an optical microscope.
[0040] 4.4 Transcriptome Analysis Transcriptome sequencing was performed on the heart tissues of the mice in the above groups. The sequencing data were filtered using SOAPnuke software (version 1.5.2). The specific filtering steps were as follows: (1) Remove reads containing sequencing adapters; (2) Remove reads with a low-quality base ratio (base quality value ≤ 5) exceeding 20%; (3) Remove reads with an unknown base ratio ("N" base) exceeding 5%. Clean reads were obtained after filtering and saved in FASTQ format. The clean reads were aligned to the reference genome using HISAT2 software (version 2.0.4); the clean reads were aligned to the reference coding gene set using Bowtie2 software (version 2.2.5), and the gene expression levels were calculated using StringTie software (version 2.1.2). Based on the gene expression in different samples, heatmaps were drawn using pheatmap software (version 1.0.8). Core differential expression analysis was performed using DESeq2 software (version 1.4.5), with a Q value ≤ 0.05 set as the screening criterion. To further explore the mechanisms of phenotypic changes, based on the hypergeometric test, enrichment analyses were conducted on the annotated differentially expressed genes using the Phyper tool (reference link: https: / / en.wikipedia.org / wiki / Hypergeometric_distribution) based on Gene Ontology (GO, reference URL: http: / / www.geneontology.org / ) and the Kyoto Encyclopedia of Genes and Genomes (KEGG, reference URL: https: / / www.kegg.jp / ). Gene set enrichment analysis (GSEA, reference URL: ...) was also performed. https: / / www.gsea-msigdb.org / gsea / index.jsp The analysis was performed using the GSEA desktop software (version 4.3.2) released by the Broad Institute. A ranking list was constructed using the normalized Log2 FoldChange values of all genes to systematically evaluate the KEGG pathway gene set in the Molecular Characteristic Database (MSigDB). The normalized enrichment score of each gene set was calculated using the phenotypic substitution test. p A value ≤ 0.05 was used for screening. The Bonferroni method was used to correct the significance level of GO terms and KEGG pathways with q-values, and a strict threshold (q-value ≤ 0.05) was set.
[0041] 4.5 Western blot (WB) detection.
[0042] Left ventricle samples from mice were used for Western blot (WB) detection of the target protein. Heart tissue samples were removed from a -80°C freezer and thawed on ice. A suitable amount of myocardial tissue was precisely cut and quickly fragmented, and protein lysis buffer was added at a ratio of 10 mg: 100 μL. The tissue was then thoroughly homogenized using a homogenizer. The lysis buffer was placed on ice and allowed to stand for 30 minutes, with gentle pipetting every 10 minutes to ensure complete lysis. The mixture was then centrifuged at 4°C and 12,000 rpm for 15 minutes, and the supernatant was carefully aspirated to obtain the tissue protein extract.
[0043] Protein concentration was determined using the BCA method. 5× protein loading buffer was added to the quantified protein sample, and the sample was heated in a 100°C metal bath for 10 minutes to fully denature the protein. 20 μg of total protein was accurately quantified for each well and loaded.
[0044] After separation by SDS-PAGE gel electrophoresis, protein bands were transferred onto a PVDF membrane. The membrane was blocked with 5% skim milk for 2 hours, followed by overnight incubation with the corresponding primary antibody at 4°C. The next day, the membrane was washed with 1×TBST and then incubated with HRP-labeled secondary antibody on a shaker at room temperature for 2 hours. After washing, chemiluminescence detection was performed using a multifunctional imaging system, and images were recorded.
[0045] Finally, the gray values of the target band were analyzed using ImageJ software, and normalized based on the expression level of the internal reference protein to calculate the relative expression level of the target protein.
[0046] 4.6 Statistical Analysis All statistical analyses and graphs were performed using GraphPad Prism software (version 10.1.2). Experimental data were compared between groups using one-way ANOVA, and results are expressed as mean ± standard deviation. p <0.05 indicates that the difference is statistically significant.
[0047] 5. Experimental Results 5.1 Cirsium glycoside improves routine parameters in HFpEF mice like Figure 1 As shown in Figure B, there was a significant difference in body weight between the normal group and the model group, but no significant difference in body weight between the drug-treated group and the model group, indicating that citronella glycoside did not significantly reduce body weight in HFpEF mice. Compared with the model group, the drug-treated group mice had significantly lower systolic blood pressure, diastolic blood pressure, and mean arterial pressure. Figure 1 As shown in CE; in addition, the results of the glucose tolerance test showed that there was no significant change in glucose tolerance in mice after administration ( Figure 1 (FG); such as Figure 1The results from the study showed that the weight of the heart and lungs, as well as the ratio of each weight to the tibia length, significantly altered cardiac hypertrophy in HFpEF mice after drug administration, but did not show a significant difference in pulmonary edema in HFpEF mice.
[0048] 5.2 Administration of citronellol improves left ventricular diastolic dysfunction in HFpEF mice like Figure 2 As shown in the middle BC diagram, there were no significant differences in ejection fraction and short-axis contraction rate among the normal group, model group, and drug-treated group; however, as Figure 2 As shown in Figures A and D, the E / E′ ratio in the treatment group was significantly lower than that in the model group; there were significant differences between the normal group and the model group in terms of end-diastolic and end-systolic interventricular septal thickness, end-diastolic and end-systolic left ventricular posterior wall thickness, and left ventricular mass, and there were also significant differences between the treatment group and the model group. Figure 2 (China EI).
[0049] 5.3 Cirsium glycosides inhibit the occurrence of myocardial misalignment, myocardial hypertrophy, and myocardial fibrosis in HFpEF mice. H&E staining results showed that the model group had disordered cardiomyocyte arrangement and inflammatory cell infiltration, and the tissue structure tended to normalize after treatment with citronellol; Masson staining results showed that the area of myocardial fibrosis in the model group was significantly increased, and the area of fibrosis was significantly reduced after treatment with citronellol; WGA staining results indicated that the cross-sectional area of cardiomyocytes in the model group was increased, and cell hypertrophy was significantly improved after treatment with citronellol. Figure 3 AC (Chinese)
[0050] 5.4 Administration of cirsinoside improves hepatic steatosis and lipid droplet distribution in HFpEF mice like Figure 4 As shown in Figure AB, liver H&E staining showed reduced vacuolation in the treatment group compared to the model group; correspondingly, the Oil Red staining area in HFpEF mice was significantly reduced after administration; at serum levels, low-density lipoprotein, total cholesterol, triglycerides, aspartate aminotransferase, and alanine aminotransferase were significantly elevated in the model group compared to the normal group, while low-density lipoprotein, total cholesterol, and alanine aminotransferase were significantly lower in the treatment group compared to the model group. Figure 4 (CG); however, in liver tissue, the expression levels of total cholesterol and triglycerides were significantly lower in the treatment group compared to the model group. Figure 4 (China HI).
[0051] 5.5 Transcriptomics reveals the potential mechanism by which piracetamine administration improves HFpEF mice Transcriptomic results showed that the top 20 differentially expressed genes between the normal group and the model group included Mzb1, Sell, CD19, Itgal, Gimap3, Gsta13, Hmgcs2, and Ptprb. Figure 5 (A); Meanwhile, a total of 131 differentially expressed genes were found between the model group and the drug-treated group, of which 51 differentially expressed genes were upregulated and 80 differentially expressed genes were downregulated. Figure 5 (B). The expression levels of Ptprb, CD19, and BNP showed significant differences between the treatment group and the model group. Specifically, Ptprb was significantly increased, while CD19 and BNP were significantly decreased. Figure 5 The top-ranked signaling pathways enriched by transcriptomics GSEA were the NF-κB signaling pathway and the PI3K / AKT signaling pathway. Figure 5 (F); Western blot analysis revealed that the expression levels of pPI3K / PI3K and pAKT / AKT were significantly higher in the treatment group compared to the model group. Figure 5 Medium GI).
[0052] 5.6 WB Results The drug piperidine can exert a therapeutic effect on HFpEF by regulating the PI3K / AKT signaling pathway.
[0053] The results of the above examples indicate that piracetamine improves the HFD+L-NAME-induced HFpEF mouse model through the PI3K / AKT signaling pathway. While having a protective effect on the heart, it also has a certain ameliorative effect on liver deterioration. Piracetamine may have a multidimensional effect on improving the function of HFpEF mice.
[0054] The above description is only a preferred embodiment of the present invention. It should be noted that for those skilled in the art, several improvements and modifications can be made without departing from the principle of the present invention, and these improvements and modifications should also be considered within the scope of protection of the present invention.
Claims
1. Application of piracetam in the preparation of drugs for the prevention and / or treatment of heart failure with preserved ejection fraction.
2. The application according to claim 1, characterized in that, The ejection fraction-preserving heart failure includes ejection fraction-preserving heart failure induced by N-nitro-L-arginine methyl ester and a high-fat diet.
3. Application of piracetam in the preparation of drugs for the prevention and / or treatment of left ventricular diastolic dysfunction caused by heart failure with preserved ejection fraction.
4. Application of citronellol in the preparation of drugs for the prevention and / or treatment of myocardial damage or cardiac hypertrophy as complications of heart failure with preserved ejection fraction.
5. The application according to claim 4, characterized in that, The myocardial damage associated with the ejection fraction-preserving heart failure includes at least one of the following conditions: cardiomyocyte disorder, myocardial hypertrophy, and myocardial fibrosis.
6. Application of citronellol in the preparation of drugs for the prevention and / or treatment of liver damage as a complication of heart failure with preserved ejection fraction.
7. The application according to claim 6, characterized in that, The liver damage associated with preservative ejection fraction heart failure includes hepatic steatosis and / or elevated blood lipid levels.
8. The application according to any one of claims 1 to 7, characterized in that, The drug includes at least one of the following dosage forms: injection, capsule, tablet, powder, granule, pill, microcapsule / microsphere formulation, suppository, ointment, spray, and targeted formulation.
9. The application according to claim 8, characterized in that, The drug also includes pharmaceutically acceptable excipients.
10. The application according to claim 9, characterized in that, The mass percentage of citronellol in the drug is 0.1% to 99.5%.