Application of colchicine in preparation of medicine for treating hypertrophic cardiomyopathy

Abstract

The invention belongs to the technical field of biological medicines, and particularly discloses application of colchicine in preparation of a medicine for treating hypertrophic cardiomyopathy. Researches show that after colchicine is used for treatment, pathological changes such as cardiac fibrosis of hypertrophic cardiomyopathy are relieved. Further experimental results show that after colchicine treatment, the proportion of pro-inflammatory macrophages in the cardiac hypertrophy tissue is reduced, the IL-6 level is reduced, and it is prompted that the anti-inflammatory ability of colchicine can improve the microenvironment of the cardiac hypertrophy tissue, so that fibrosis is relieved, and the cardiac function is improved. The invention proves the clinical transformation potential of colchicine in the treatment of cardiac hypertrophy patients. If the medicine is applied clinically in the future, the risk of sudden cardiac death is expected to be reduced by relieving cardiac fibrosis, and a new treatment choice is provided for improving the prognosis of a hypertrophic cardiomyopathy patient.

Description

Technical Field

[0001] This invention belongs to the field of biomedical technology, specifically relating to the application of colchicine in the preparation of drugs for treating hypertrophic cardiomyopathy. Background Technology

[0002] Hypertrophic cardiomyopathy (HCM) is the most common inherited heart disease, with a prevalence of approximately 1 in 500. Characterized by left ventricular hypertrophy and diastolic dysfunction, HCM histologically presents as cardiomyocyte hypertrophy, disordered arrangement, and myocardial fibrosis. It is a leading cause of sudden cardiac death, progressive heart failure, and other malignant cardiovascular events, especially in young people; however, the mechanisms of hypertrophic cardiomyopathy-related arrhythmias remain incompletely understood.

[0003] Current research indicates that while sarcomere gene mutations and their effects can induce myocardial hypertrophy, other phenotypic features associated with myocardial hypertrophy, such as sudden cardiac death, are actually driven by downstream events. One characteristic of hypertrophic cardiomyopathy is progressive tissue remodeling accompanied by significant fibrosis, and myocardial scar burden measured by magnetic resonance imaging can serve as a strong predictor of adverse patient outcomes. Clarifying the driving mechanisms of myocardial tissue remodeling in hypertrophic cardiomyopathy will lay a solid foundation for the future development of precision prevention and treatment strategies.

[0004] Current treatments for myocardial hypertrophy, including allosteric myosin modulators, only provide temporary relief from cardiac dysfunction. Without addressing or reversing fibrosis, focusing solely on functional improvements within the disease has very limited long-term therapeutic effects. Current research has identified immune activation and inflammatory responses as key drivers of myocardial hypertrophy progression. Hypertrophic myocardial tissue triggers an inflammatory signaling cascade, inducing immune cell infiltration, cytokine (TNF-α, IL-6, IL-1β) release, and fibrosis, ultimately leading to myocardial remodeling. Macrophages play a central regulatory role in this process; under specific conditions, pro-fibrotic and pro-inflammatory macrophages can infiltrate myocardial tissue, exacerbating cardiac dysfunction.

[0005] Colchicine, a classic anti-inflammatory drug, has shown in recent studies to exert pleiotropic effects on neutrophils, endothelial cells, and macrophages by inhibiting microtubule polymerization and NLRP3 inflammasome assembly in myocardial tissue. This, in turn, inhibits the production of key pro-inflammatory factors such as IL-1β and IL-6. Colchicine is primarily used to treat acute gout, rapidly relieving joint pain caused by gout by suppressing inflammatory responses and reducing leukocyte chemotaxis. It can also effectively reduce the risk of adverse cardiovascular events in patients with coronary syndrome. However, colchicine has not yet been used to treat hypertrophic cardiomyopathy. Summary of the Invention

[0006] The study of this invention shows that colchicine can reduce myocardial inflammation and fibrosis in hypertrophic cardiomyopathy, suggesting that it has excellent clinical translation potential and may be applied in the future to reduce cardiac fibrosis, reduce the risk of sudden death, and thus benefit more patients with hypertrophic cardiomyopathy.

[0007] The specific technical solution adopted in this invention is as follows:

[0008] This invention provides the use of colchicine in the preparation of a medicament for treating hypertrophic cardiomyopathy or preventing malignant cardiovascular diseases caused by hypertrophic cardiomyopathy, wherein the medicament uses colchicine as the active ingredient.

[0009] Furthermore, the colchicine can alleviate myocardial inflammation and fibrosis in hypertrophic cardiomyopathy.

[0010] Furthermore, the colchicine reduces the proportion of pro-inflammatory macrophages and decreases IL-6 levels in hypertrophic myocardial tissue, suggesting its anti-inflammatory ability. It can improve the microenvironment of hypertrophic myocardial tissue, thereby reducing fibrosis and improving cardiac function.

[0011] Furthermore, the colchicine downregulates the mRNA expression levels of the myocardial hypertrophy marker genes Anp and Bnp, as well as the fibrosis-related genes Ctgf, Col1a1, and Col3a1.

[0012] Furthermore, the specific manifestations of improved cardiac function include: improved ejection fraction and fractional shortening, reduced heart weight to body weight ratio, reduced cardiomyocyte size, and reduced area of ​​cardiac interstitial fibrosis.

[0013] Furthermore, the aforementioned malignant cardiovascular diseases include sudden cardiac death and progressive heart failure.

[0014] Furthermore, the drug may also include pharmaceutically acceptable excipients. "Pharmaceutically acceptable" means that when properly administered to animals or humans, the excipients do not produce adverse, allergic, or other adverse reactions. The pharmaceutically acceptable excipients may be selected from one or more of diluents, fillers, surfactants, absorption enhancers, disintegrants, wetting agents, dispersants, etc.

[0015] Furthermore, the dosage form of the drug can be selected from any one of injections, emulsions, tablets, powders, granules, gels, ointments, capsules, oral liquids, etc.

[0016] The present invention has the following beneficial effects:

[0017] This invention constructed two heterozygous mouse models carrying gene mutations associated with the risk of sudden cardiac death. Single-cell sequencing revealed fibrosis and immune activation phenotypes in hypertrophic myocardial tissue. Furthermore, colchicine treatment alleviated pathological changes such as cardiac fibrosis. Further experimental results showed that colchicine treatment reduced the proportion of pro-inflammatory macrophages and decreased IL-6 levels in hypertrophic myocardial tissue, suggesting that colchicine's anti-inflammatory ability can improve the microenvironment of hypertrophic myocardial tissue, thereby reducing fibrosis and improving cardiac function. This invention demonstrates the clinical translational potential of colchicine in the treatment of patients with hypertrophic myocardial disease. If this drug is applied clinically in the future, it is expected to reduce the risk of sudden cardiac death by alleviating cardiac fibrosis, providing a new treatment option for improving the prognosis of patients with hypertrophic cardiomyopathy. Attached Figure Description

[0018] Figure 1 In Example 1, the Prkag2 p.R299Q and Myh6 p.R404Q mouse models reproduced key pathological features of hypertrophic cardiomyopathy (HCM). Figures show: A. Schematic diagram of HCM model construction using CRISPR / Cas9 technology. B. Left ventricular posterior wall thickness analysis using two-dimensional mouse echocardiography (n=9 / group). *P < 0.001, ****P < 0.001. C. Calculation of heart weight to body weight ratio (n=6 / group). P < 0.01. D. Representative images of wheat germ lectin staining of the heart in each group. The right side shows the quantitative analysis of cardiomyocyte size. Scale bar = 50 μm. **P < 0.001. E. Representative images of Masson's trichrome staining of the heart in each group. The right side shows the quantitative analysis of cardiac interstitial fibrosis area (n=9 / group). Scale bar = 100 μm. P < 0.001. F and G. Relative expression levels of Anp (atrial natriuretic peptide) and Bnp (brain natriuretic peptide) mRNA in mouse heart tissue (n=6 / group) *P <0.05, P < 0.01, *P < 0.001 (F), and relative expression levels of Ctgf (connective tissue growth factor), Col1a1 (type I collagen α1 chain), and Col3a1 (type III collagen α1 chain) (G). Data are expressed as mean ± standard error and were analyzed by one-way ANOVA (D, E) and unpaired t-tests (F, G).

[0019] Figure 2Example 2: Single-cell RNA sequencing revealed cardiac fibrosis and immune microenvironment activation in HCM mice. Figures show: A. Representative functional entries of significantly upregulated genes in heart cells of HCM and wild-type mice; corrected P < 0.05. B. Heatmap showing representative upregulated genes related to fibrosis in fibroblasts; corrected P < 0.05. C. Gene set enrichment analysis of pathway activation in HCM mouse fibroblasts (P < 0.05). D. UMAP showing fibroblast subsets. E. Relative proportions of each subset in fibroblasts from different groups. F. Bubble plot showing distinct characteristics of the FB4 subset. G. UMAP showing immune cell clusters. I. Relative proportions of each cluster in immune cells from different groups. H. Dot plot showing molecular characteristics of each immune cell type. J. Gene set enrichment analysis of pathway activity in macrophages of HCM mice (P < 0.05). Representative functional entries of significantly upregulated genes in cardiac macrophages of K. HCM mice and wild-type mice; corrected P value < 0.05.

[0020] Figure 3 Example 3: Colchicine treatment restored cardiac function and reduced cardiac fibrosis in HCM mice. Figure: A. Quantification of cardiac function (ejection fraction and fractional shortening) after 4 weeks of colchicine treatment using two-dimensional mouse echocardiography *P<0.05. B. Heart weight to body weight ratio calculated at the experimental endpoint (n_Myh6 group=7, n_Prkag2 group=8) *P <0.05. C. Representative images of wheat germ lectin staining in the hearts of each group. The right side shows the quantitative analysis of cardiomyocyte size. Scale bar = 50μm (n_Myh6 group=6, n_Prkag2 group=7) P < 0.01, *P < 0.001. D. Representative images of Masson's trichrome staining in the hearts of each group. The right side shows the quantitative analysis of the area of ​​cardiac interstitial fibrosis. Scale bar = 100 μm (n_Myh6=5, n_Myh6-Col=5, n_Prkag2=4, n_Prkag2-Col=5) P < 0.01, *P < 0.001. E and F. The experimental endpoint was the detection of the relative expression levels of Anp mRNA and Bnp mRNA in mouse heart tissue (n_Myh6 group = 6, n_Prkag2 group = 4) P < 0.05, P < 0.01, P < 0.001 (E), and the relative expression levels of Ctgf, Col1a1, and Col3a1 (F). Data are expressed as mean ± standard error and were statistically analyzed using one-way ANOVA (CF).

[0021] Figure 4Example 4: Colchicine treatment reduced pro-inflammatory macrophages and decreased IL-6 production in M1 macrophages. Figure: A. Flow cytometry analysis of M1 macrophage frequency and mean fluorescence intensity in cardiac tissue, showing representative flow cytometry patterns and statistical results (n=9) **P < 0.01. B. Detection of relative IL-6 mRNA expression in mouse cardiac tissue at the experimental endpoint (n_Myh6 group=6, n_Prkag2 group=4) P < 0.05, **P < 0.001, ****P < 0.001. C. Detection of IL-6 protein level in mouse cardiac tissue at the experimental endpoint (n_Myh6 group=6, n_Prkag2 group=4) *P < 0.05, **P < 0.01, ****P < 0.001. D. AUCell analysis showing the activity score of the IL-6 production pathway in different cardiac cell populations. E. IL-6 levels in the supernatant of M1 polarized macrophages after colchicine treatment were detected by ELISA (n=3). P < 0.05, P < 0.01, P < 0.001, ****P < 0.001. Detailed Implementation

[0022] To verify the role of colchicine in myocardial hypertrophy, we selected two representative pathogenic sites identified in patients with hypertrophic cardiomyopathy, which carry a high risk of sudden cardiac death: the classic sarcomere protein Myh6 p.R404Q mutation and the non-sarcomere protein Prkag2 p.R299Q mutation. Among the numerous sarcomere gene mutations associated with hypertrophic cardiomyopathy, Myh6 mutations account for more than one-third of all cases, disrupting sarcomere contractile function and significantly associated with the risk of sudden cardiac death. Patients carrying the Prkag2 mutation develop Prkag2 syndrome, characterized by myocardial hypertrophy, ventricular pre-excitation, and conduction abnormalities, thereby increasing their risk of sudden cardiac death. We constructed heterozygous mouse models carrying the two aforementioned mutations to simulate sarcomere-related and metabolism-related myocardial hypertrophy, respectively. Using these two models, we aim to elucidate whether colchicine can alleviate myocardial hypertrophy and improve cardiac structure and function by regulating the cardiac immune microenvironment and alleviating cardiac fibrosis, thus providing preclinical evidence for colchicine treatment of patients with hypertrophic cardiomyopathy.

[0023] Hypertrophic cardiomyopathy (HCM) is characterized by left ventricular hypertrophy and diastolic dysfunction. Histologically, it manifests as cardiomyocyte hypertrophy, disordered arrangement, and myocardial fibrosis, and is a major cause of sudden cardiac death and other malignant cardiovascular events. One of the defining features of HCM is progressive myocardial remodeling accompanied by significant fibrosis. Previous studies have shown that myocardial scar burden measured by magnetic resonance imaging (MRI) can serve as a strong predictor of adverse cardiovascular events. The 2020 American Heart Association (AHA) guidelines listed delayed gadolinium enhancement on cardiac MRI as a clinical risk factor for sudden cardiac death in patients with hypertrophic cardiomyopathy. In this study, we constructed two representative heterozygous mouse models with a high risk of sudden cardiac death. Single-cell sequencing revealed the presence of fibrosis and immune activation phenotypes in hypertrophic myocardial tissue. After colchicine treatment, the pathological changes of myocardial hypertrophy, such as myocardial fibrosis, were alleviated. This study demonstrates that colchicine can reduce inflammation and fibrosis in hypertrophic myocardial tissue and may be applied to patients with hypertrophic myocardial disease to reduce cardiac remodeling and lower the risk of sudden cardiac death.

[0024] In normal myocardial tissue, cardiac conduction does not re-enter within ventricular cells. However, in patients with myocardial hypertrophy, due to myocardial fibrosis and other factors, unidirectional blockage may occur during conduction, forming a re-entry loop and leading to rapid ventricular arrhythmias. Furthermore, myocardial fibrosis disrupts cardiac structure, impairs excitation-contraction coupling, and affects cardiac systolic and diastolic function. The core processes of myocardial fibrosis include the transformation of fibroblasts into myofibroblasts and are related to inflammatory factors released by immune cells, such as TGF-β, TNF-α, and interleukins. Researchers have used single-cell sequencing technology to discover cell communication between immune cells and fibroblasts in the hearts of heart failure patients, highlighting the therapeutic potential of targeting inflammation to treat myocardial fibrosis and protect cardiac function. Our scRNA-seq results in a mouse model also showed upregulation of immune response-related pathways in hypertrophic myocardial tissue. The significantly altered immune microenvironment in hypertrophic myocardial tissue may provide new therapeutic avenues for disease intervention.

[0025] Since the immune microenvironment is closely related to myocardial fibrosis, we hope to reduce myocardial fibrosis and improve cardiac function by improving the immune microenvironment of hypertrophic myocardial tissue, thereby reducing the risk of sudden cardiac death in patients with hypertrophic myocardial disease. Colchicine, as an anti-inflammatory drug, has shown advantages in the treatment of various cardiovascular diseases in recent years. However, its role in the treatment of hypertrophic myocardial disease remains unclear. Our experimental results show that in a mouse model of hypertrophic myocardial disease, the anti-inflammatory drug colchicine can alleviate pathological changes, including fibrosis, by reducing pro-inflammatory macrophages and lowering IL-6 levels.

[0026] Our findings confirm that colchicine can reduce pro-inflammatory macrophages in myocardial tissue and improve the myocardial microenvironment. In fact, macrophages, as the first immune cells to respond to injury, play a crucial role in the myocardial microenvironment. Traditionally, macrophage polarization has been classified into M1-like and M2-like phenotypes: the former exhibits pro-inflammatory properties, while the latter displays anti-inflammatory and repair phenotypes. For example, the hypoglycemic drug dapagliflozin can effectively alleviate myocardial fibrosis after myocardial infarction by inhibiting macrophage inflammatory pathways and promoting their repair function. Our study found that pro-inflammatory macrophages are involved in the process of myocardial hypertrophy, suggesting that therapeutically reducing these cells may delay disease progression.

[0027] Researchers using single-cell sequencing have discovered a unique group of cardiac-resident macrophages that can prevent cardiac fibrosis, highlighting the significance of utilizing cutting-edge single-cell technology to explore cardiac immune-fibrotic interactions. In our single-cell sequencing results, macrophages from hypertrophic myocardial mice exhibited a significantly proliferative phenotype compared to wild-type mice, allowing for further investigation into the mechanisms driving their proliferation / activation and the origin of these proliferating macrophages. Furthermore, we identified a fibroblast subset with immunomodulatory functions, whose roles in myocardial tissue repair, maintenance of microenvironmental homeostasis, and interaction with immune components can be further investigated. These findings will deepen our understanding of the microenvironment in hypertrophic myocardial tissue and may lead to novel therapeutic strategies targeting these mechanisms. Future research needs to leverage cutting-edge single-cell technology to define specific cell subsets and reveal their spatiotemporal dynamics and functional roles. Besides macrophages, other immune cell populations also play important roles in shaping the immune microenvironment in hypertrophic myocardial tissue. Future research should further explore more immune cell subsets and their intercellular interactions to construct a more complete cardiac immune atlas and deepen the understanding of immune-fibrotic changes in hypertrophic myocardial tissue.

[0028] The present invention will be further described below with reference to the accompanying drawings and specific embodiments. Unless otherwise specified, the reagents used in the following embodiments are all commercially available conventional reagents, and the experimental procedures involved are all conventional procedures in the art unless otherwise specified.

[0029] Example 1: Construction and Phenotypic Validation of Myh6 and Prkag2 Mutant Mouse Models

[0030] This embodiment constructed heterozygous mouse models carrying two mutations, Myh6 p.R404Q and Prkag2 p.R299Q, to simulate sarcomere-related and metabolism-related myocardial hypertrophy, respectively, and verified the successful construction of the two hypertrophic cardiomyopathy models, as detailed below:

[0031] Experimental methods: such as Figure 1As shown in Figure A, heterozygous mutant mice of Myh6 p.R404Q (hereinafter referred to as Myh6) and Prkag2 p.R299Q (hereinafter referred to as Prk) were constructed using CRISPR / Cas9 technology. Wild-type mice were used as controls (WT). The following tests were performed on the constructed Myh6 and Prk mutant models: Left ventricular posterior wall thickness was analyzed using two-dimensional mouse echocardiography. The heart weight to body weight ratio was calculated. Heart tissue was stained with wheat germ lectin and Masson's trichrome staining. Cardiac cell size and the area of ​​cardiac interstitial fibrosis were observed and quantified under a microscope. RNA was extracted from myocardial tissue, and the expression levels of myocardial hypertrophy marker genes (Anp, Bnp) and fibrosis-related genes (Ctgf, Col1a1, Col3a1) were detected using qPCR.

[0032] Experimental results: Figure 1 B-mode echocardiographic analysis showed a significant increase in the thickness of the left ventricular posterior wall in both models; Figure 1 C shows that the ratio of heart weight to body weight is significantly increased in both models; Figure 1 The wheat germ lectin staining results of D showed that the size of cardiomyocytes was significantly increased in both models; Figure 1 The results of E's Masson trichrome staining showed a significant increase in the area of ​​cardiac interstitial fibrosis in both models; Figure 1 qPCR results from FG confirmed that the expression of hypertrophic cardiomyopathy marker genes Anp and Bnp, as well as fibrosis-related genes Ctgf, Col1a1, and Col3a1, was significantly upregulated in both models. These results indicate that the Myh6 and Prk mouse models reproduce the key pathological features of HCM, and this embodiment successfully constructed two hypertrophic cardiomyopathy mouse models, Myh6 and Prk.

[0033] Example 2: Single-cell sequencing analysis of the immune microenvironment in myocardial tissue

[0034] This embodiment reveals fibrosis and immune activation phenotypes in hypertrophic myocardial tissue through single-cell sequencing of constructed Myh6 and Prk mouse models. Details are as follows:

[0035] Experimental Methods: Single-cell RNA sequencing (scRNA-seq) was performed on heart tissues from Myh6 and Prk mutant mouse models and wild-type control mice. Heart cells were isolated, single-cell suspensions were prepared, libraries were constructed using the 10x Genomics platform, and sequencing was performed. Sequencing data underwent quality control, normalization, and cluster analysis to identify different cell populations, including fibroblast subsets and immune cell clusters. Gene set enrichment analysis (GSEA) and differentially expressed gene analysis were performed to assess the expression of fibrosis-related genes and the activation status of immune pathways.

[0036] Experimental results:

[0037] Regarding fibrosis, Figure 2 A showed that genes significantly upregulated in the cardiac cells of HCM mice were enriched in functional entries related to fibrosis and immune activation. Figure 2 The heatmap of B shows representative upregulated genes associated with fibrosis in fibroblasts; Figure 2 The GSEA results of C showed that the collagen trimer pathway was significantly activated in HCM mouse fibroblasts compared with wild-type controls; Figure 2 The UMAP plot of D identified four fibroblast subsets; Figure 2 E shows the relative proportions of different subpopulations in fibroblasts from different groups; Figure 2 The bubble chart of F shows that the FB4 subgroup has distinct features.

[0038] To investigate changes in the immune pattern in hypertrophic hearts, we further subdivided the types of immune cells. Figure 2 G shows that macrophages constitute the largest subset of immune cells in the dataset; Figure 2 The H-plot illustrates the molecular characteristics of various immune cell types, identifying different cell clusters; we quantified the changes in immune cell composition. Figure 2 I showed that the proportion of macrophage 5 (proliferating macrophages), characterized by high expression of proliferation markers (Mki67, Top2a, and Neil3), was significantly increased in the HCM mouse model compared to the wild-type control group. Furthermore, Figure 2 JK showed that cardiac macrophages in hypertrophic hearts exhibited an activated phenotype, with significant enrichment of proliferation-related and immune-related pathways. Notably, these phenotypic features showed high conservation in both HCM models, suggesting that the major sarcoprotein-related subtypes and the rare non-sarcoprotein-related HCM subtypes may share some commonalities in the immune microenvironment.

[0039] In summary, single-cell RNA sequencing data revealed the simultaneous presence of fibrosis and activation of the immune microenvironment in the heart of HCM mice. These changes involved multiple cell types, with fibroblasts and macrophages being particularly prominent.

[0040] Example 3: Improvement of myocardial tissue pathology after colchicine treatment

[0041] This embodiment evaluated the therapeutic effects of colchicine on cardiac function, myocardial hypertrophy, and fibrosis in HCM mice, as detailed below:

[0042] Experimental Methods: Myh6 and Prk mutant mouse models were treated with colchicine (purchased from MedChemExpress, Shanghai, China) for 4 weeks: 0.2 mg / kg daily, administered by gavage for 30 consecutive days. After treatment, cardiac function was assessed using two-dimensional echocardiography, the heart weight to body weight ratio was calculated, cardiomyocyte size was assessed by wheat germ lectin staining, the area of ​​cardiac interstitial fibrosis was assessed by Masson's trichrome staining, and the mRNA expression levels of Anp, Bnp, Ctgf, Col1a1, and Col3a1 were detected by qPCR. Experimental groups included: wild-type control group (WT), model control group (Myh6 / Prk), and model treatment group (Myh6-Col / Prk-Col).

[0043] Experimental results: Figure 3 A showed that after 4 weeks of colchicine treatment, compared with the model control group, the ejection fraction and short axis shortening rate of mice in the model treatment group were significantly improved. Figure 3 B shows that the ratio of heart weight to body weight was significantly lower in the treatment group compared to the control group. Figure 3 C's wheat germ lectin staining showed that the size of cardiomyocytes in the model treatment group was significantly smaller than that in the model control group; Figure 3 Masson's trichrome staining showed that the area of ​​cardiac interstitial fibrosis was significantly reduced in the treatment group compared with the control group. Figure 3 qPCR analysis of EF showed that the expression levels of hypertrophic cardiomyopathy marker genes Anp and Bnp, as well as fibrosis-related genes Ctgf, Col1a1, and Col3a1, were significantly downregulated in the treatment group compared to the control group. These results indicate that colchicine treatment can restore cardiac function and reduce cardiac fibrosis in HCM mice, effectively treating hypertrophic cardiomyopathy.

[0044] Example 4: Colchicine inhibits IL-6 expression and macrophage activation

[0045] This embodiment elucidates the specific mechanism by which colchicine regulates macrophage function and IL-6 expression, as follows:

[0046] Experimental methods: Heart tissue was collected from Myh6 and Prkag2 mutant mouse models and mice in the colchicine treatment group at the experimental endpoint. Flow cytometry was used to analyze the frequency and average fluorescence intensity of M1 macrophages in the heart tissue. qPCR was used to detect the relative expression level of IL-6 mRNA, and ELISA was used to detect the IL-6 protein level. AUCell analysis was performed on scRNA-seq data to evaluate the activity score of the IL-6 production pathway in different cell populations of the heart. M1 polarized macrophages were cultured in vitro and the supernatant was collected after treatment with different concentrations of colchicine. IL-6 level was detected by ELISA.

[0047] Experimental results: Figure 4Flow cytometry analysis of A showed that the frequency and mean fluorescence intensity of M1 macrophages in cardiac tissue were significantly reduced after colchicine treatment. Figure 4 qPCR analysis of B showed that IL-6 mRNA expression was significantly downregulated; Figure 4 ELISA analysis of C showed a significant decrease in IL-6 protein levels; Figure 4 AUCell analysis of D showed that colchicine treatment reduced the activity score of the IL-6 production pathway in various cardiac cell populations. Figure 4 In vitro experiments with E confirmed that colchicine treatment significantly reduced the IL-6 level in the supernatant of M1 polarized macrophages. These results indicate that colchicine treatment can reduce pro-inflammatory macrophages and inhibit the expression of the pro-inflammatory cytokine IL-6.

[0048] This specific embodiment is merely an explanation of the present invention and is not intended to limit the present invention. Any changes made by those skilled in the art after reading the specification of the present invention, as long as they are within the scope of the claims of the present invention, will be protected by patent law.

Claims

1. The use of colchicine in the preparation of drugs for the treatment of hypertrophic cardiomyopathy or the prevention of malignant cardiovascular diseases caused by hypertrophic cardiomyopathy, wherein the drugs use colchicine as the active ingredient.

2. The application according to claim 1, characterized in that, The colchicine can reduce myocardial inflammation and fibrosis in hypertrophic cardiomyopathy.

3. The application according to claim 2, characterized in that, The colchicine reduces the proportion of pro-inflammatory macrophages and decreases IL-6 levels in hypertrophic myocardial tissue, suggesting its anti-inflammatory ability. It can improve the microenvironment of hypertrophic myocardial tissue, thereby reducing fibrosis and improving cardiac function.

4. The application according to claim 2, characterized in that, The colchicine downregulated the mRNA expression levels of the myocardial hypertrophy marker genes Anp and Bnp, as well as the fibrosis-related genes Ctgf, Col1a1, and Col3a1.

5. The application according to claim 3, characterized in that, The specific manifestations of improved cardiac function include: improved ejection fraction and fractional shortening, reduced heart weight to body weight ratio, reduced cardiomyocyte size, and reduced area of ​​cardiac interstitial fibrosis.

6. The application according to claim 1, characterized in that, The aforementioned malignant cardiovascular diseases include sudden cardiac death and progressive heart failure.

7. The application according to claim 1, characterized in that, The drug also includes pharmaceutically acceptable excipients.

8. The application according to claim 7, characterized in that, The pharmaceutically acceptable excipients are selected from one or more of the following: diluents, fillers, surfactants, absorption enhancers, disintegrants, wetting agents, and dispersants.

9. The application according to claim 1, characterized in that, The dosage form of the drug is any one of injection, emulsion, tablet, powder, granule, gel, ointment, capsule, or oral liquid.

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