Biomarker for diagnosis and prognosis evaluation of ventricular arrhythmia of dilated cardiomyopathy and application of biomarker

Abstract

The invention relates to the field of medicine, in particular to a biomarker for diagnosis and prognosis evaluation of ventricular arrhythmia of a patient with dilated cardiomyopathy (DCM) and application of the biomarker. Matrix metalloproteinase-9 (MMP-9), 5-hydroxytryptamine (5-HT), noradrenaline (NE) and acetylcholine (ACh) are screened to serve as a biomarker combination, and by measuring the levels of MMP-9, 5-HT, NE and ACh in plasma, whether a DCM patient is accompanied with ventricular arrhythmia or not can be effectively recognized, the severity of the condition of the patient is evaluated, and the prognosis risk of the patient is predicted. The synergistic effect of MMP-9, 5-HT, NE and ACh can complement the defects of a single index, the diagnosis efficiency is improved, particularly, high specificity is achieved for ventricular arrhythmia, and an important basis is provided for risk stratification and individualized treatment of DCM patients.

Description

Technical Field

[0001] This invention belongs to the medical field, and specifically relates to biomarkers for the diagnosis and prognostic assessment of ventricular arrhythmias in dilated cardiomyopathy and their applications. Background Technology

[0002] Dilated cardiomyopathy (DCM) is a primary cardiomyopathy characterized by progressive decline in ventricular dilation and systolic function, with a global incidence of approximately 1 in 2,500. Clinical data shows that approximately 40%-60% of DCM patients develop ventricular arrhythmias, with an annual incidence of malignant ventricular arrhythmias of 2.4%-5.6%, significantly increasing the risk of sudden cardiac death. In recent years, with the development of medical imaging and molecular biology techniques, significant progress has been made in the diagnosis and risk stratification of DCM, but early diagnosis, disease assessment, and prognosis prediction still face challenges. In particular, existing methods for assessing the risk of ventricular arrhythmias, such as electrophysiological examinations, cardiac magnetic resonance imaging, and electrocardiogram parameters, have certain limitations and fail to meet the needs of individualized clinical risk assessment and precision medicine.

[0003] Biomarkers, as objective and quantifiable indicators reflecting specific pathophysiological processes, play an increasingly important role in the diagnosis and risk assessment of cardiovascular diseases. Traditional biomarkers for myocardial injury and heart failure, such as cardiac troponin (cTn), brain natriuretic peptide (BNP), and N-terminal pro-Brain natriuretic peptide (NT-proBNP), are widely used in clinical practice (heart failure assessment). However, these indicators have poor specificity for ventricular arrhythmias and limited value in assessing diabetic cerebrovascular disease (DCM), especially in high-risk patients with ventricular arrhythmias. Therefore, exploring novel biomarkers to improve the accuracy of DCM diagnosis and the precision of patient risk stratification is of significant clinical importance.

[0004] Therefore, finding more effective biomarkers is crucial for improving the accuracy of early diagnosis of ventricular arrhythmias in patients with DCM, assessing disease severity, and improving patient prognosis. Summary of the Invention

[0005] This invention aims to provide biomarkers for the diagnosis and prognostic assessment of ventricular arrhythmias in dilated cardiomyopathy, and to screen for combinations of biomarkers that can predict the occurrence and severity of ventricular arrhythmias earlier and more accurately, especially for differentiating high-risk patients, and have positive significance for prognostic assessment and treatment adjustment.

[0006] On the one hand, the present invention provides biomarkers for the diagnosis and prognostic assessment of ventricular arrhythmias in dilated cardiomyopathy, including combinations of MMP-9, 5-HT, NE and ACh.

[0007] On the other hand, the present invention also provides the use of products that detect MMP-9, 5-HT, NE and / or ACh in the preparation of tools for diagnosing ventricular arrhythmias in dilated cardiomyopathy or predicting its prognosis.

[0008] Furthermore, the products for detecting MMP-9, 5-HT, NE, or / and ACh include products for detecting their expression levels, such as ELISA kits.

[0009] Furthermore, the product was used to measure the levels of MMP-9, 5-HT, NE, and ACh in the subject's sample. Compared with normal individuals, the expression levels of MMP-9, 5-HT, and NE in the subject's sample were increased, while the expression level of ACh was decreased, which suggests that the subject has ventricular arrhythmia or a poor prognosis.

[0010] Furthermore, the product is used to measure the levels of MMP-9, 5-HT, NE, and ACh in subject samples and compare these levels with predetermined thresholds for the diagnosis and risk stratification of DCM with ventricular arrhythmias.

[0011] Furthermore, the subject samples were derived from plasma.

[0012] In another aspect, the present invention provides a tool for diagnosing ventricular arrhythmias in dilated cardiomyopathy or predicting its prognosis. The tool includes a data analysis module and reagents for detecting biomarkers MMP-9, 5-HT, NE, and ACh. The data analysis module is used to analyze the detection values ​​of biomarkers in subject samples.

[0013] Furthermore, compared with normal individuals, when the levels of MMP-9, 5-HT, and / or NE were elevated and the level of ACh was decreased, it was considered that the subjects had ventricular arrhythmias. MMP-9, 5-HT, and NE were positively correlated with the degree of cardiac structural and functional damage, while ACh was negatively correlated with the degree of cardiac structural and functional damage. When the MMP-9 level is ≥16.85 ng / mL, 5-HT is ≥756.32 ng / mL, NE is ≥85.46 ng / L, and ACh is ≤28.45 ng / mL, the subject is considered to have dilated cardiomyopathy with high-risk ventricular arrhythmia.

[0014] Furthermore, the biomarkers also include echocardiographic parameters, including LVEDD, LVESD, LVEF, LVFS, E / A ratio, and Tei index, which are used in conjunction with MMP-9, 5-HT, NE, and ACh to assess the risk of ventricular arrhythmias and cardiac function status.

[0015] Furthermore, the subject samples were derived from plasma.

[0016] Beneficial technical effects of the present invention: This study is the first to systematically reveal the characteristic alterations and synergistic mechanisms of four biomarkers—MMP-9, 5-HT, NE, and ACh—in patients with dilated cardiomyopathy (DCM) and ventricular arrhythmias (VA). It has the following clinical value and significance: A novel combination of biomarkers: MMP-9, 5-HT, NE, and ACh provides a novel and efficient tool for the early identification, risk stratification, and prognostic assessment of VA in DCM patients.

[0017] Potential therapeutic targets: This study suggests that a combined intervention strategy targeting MMP-9 (matrix remodeling), the 5-HT system (neuroendocrine), and autonomic balance (NE / ACh) may become a new approach for the prevention and treatment of DCM-related VA.

[0018] Improved diagnostic efficacy: The synergistic effect of MMP-9, 5-HT, NE and ACh can complement the shortcomings of a single indicator and improve diagnostic efficacy, especially for ventricular arrhythmias, which have high specificity and can help develop more accurate treatment plans for patients.

[0019] By jointly measuring the levels of MMP-9, 5-HT, NE, and ACh in a patient's plasma and comparing these levels with predetermined risk thresholds, the risk of ventricular arrhythmias and prognosis can be predicted. Attached Figure Description

[0020] Figure 1 This study compares plasma biochemical parameters between the DCM with ventricular arrhythmia group and the DCM alone group.

[0021] Figure 2 To compare echocardiographic parameters between the two groups of patients.

[0022] Figure 3 Correlation analysis of plasma biochemical indicators and echocardiographic parameters. Detailed Implementation

[0023] The following detailed description illustrates the specific implementation method: 1. Research Subjects 1.1 Sample Collection and Grouping Seventy-nine patients with dilated cardiomyopathy were included and divided into a study group (DCM with ventricular arrhythmia, n=41) and a control group (DCM alone, n=38) based on whether they had ventricular arrhythmias. This study has been ethically approved (approval number: NM-LL-2025-01-17-83), and all participants have signed informed consent forms.

[0024] Inclusion criteria included: (1) age 30-80 years, gender not limited; (2) meeting the diagnostic criteria for DCM in the 2018 Chinese Guidelines for the Diagnosis and Treatment of Dilated Cardiomyopathy; (3) patients in the study group also meeting the diagnostic criteria for ventricular arrhythmia in the 2020 Chinese Expert Consensus on Ventricular Arrhythmias; (4) NYHA functional classification of I-IV; (5) patients with normal cognitive function and able to cooperate in completing relevant examinations. Exclusion criteria included: patients with hematologic disorders or malignancies, severe infectious diseases, metabolic diseases, endocrine disorders, mental disorders, and those who had used antibiotics, glucocorticoids, or immunosuppressants within the past two months.

[0025] 1.2 Detection Method 1.2.1 Detection of plasma MMP-9, 5-HT, NE, and ACh: ELISA method (Shanghai Langdun Biotechnology Reagent Kit).

[0026] Sample preparation: Collect peripheral venous blood, anticoagulate with EDTA, centrifuge to separate plasma, and store at -80℃.

[0027] Detection steps: Follow the instructions in the kit's manual.

[0028] 1.2.2 Echocardiographic examination: Philips EPIQ7C color Doppler ultrasound diagnostic instrument was used to measure LVEDD, LVESD, LVEF, LVFS, E / A ratio and Tei index.

[0029] 2. Key Data 2.1 Comparison of baseline data between the two groups of patients There were no statistically significant differences between the study group (n=41) and the control group (n=38) in terms of age, gender composition, body mass index (BMI), disease duration, NYHA functional classification, and basic medication (all P>0.05). See Table 1.

[0030] Table 1. Comparison of baseline clinical data between the two groups of patients [x̄±s, n(%)]

[0031] 2.2 Comparison of plasma biochemical indicators between the two groups of patients The levels of MMP-9, 5-HT, and NE in the study group were significantly higher than those in the control group, while the level of ACh was significantly lower than that in the control group (P<0.001).

[0032] Compared with the control group, the plasma MMP-9, 5-HT, and NE levels in the study group were significantly increased (16.85±4.23 vs 12.37±3.86 ng / ml, 756.32±142.58 vs 583.64±128.95 ng / ml, 85.46±18.73 vs 62.35±15.84 ng / L, all P<0.001), while ACh levels were significantly decreased (28.45±6.92 vs 38.64±8.15 ng / ml, t=6.152, P<0.001). See Table 2. Figure 1 .

[0033] Table 2 Comparison of plasma biochemical indicators between the two groups of patients (x̄±s)

[0034] 2.3 Comparison of echocardiographic parameters between the two groups of patients The study group had significantly higher LVEDD, LVESD, E / A ratio, and Tei index than the control group, while LVEF and LVFS were significantly lower in the study group (P<0.001).

[0035] Compared with the control group, the left ventricular end-diastolic diameter (LVEDD) and end-systolic diameter (LVESD) of the patients in the study group were significantly increased (68.42±7.85 vs 53.26±4.37 mm, 58.96±7.23 vs 35.74±4.85 mm, both P<0.001), while the left ventricular ejection fraction (LVEF) and fractional shortening (LVFS) were significantly decreased (27.35±6.42% vs 59.86±4.78%, 13.62±3.84% vs 32.58±3.95%, both P<0.001). Meanwhile, the E / A ratio in the study group was significantly increased (2.46±0.68 vs 1.28±0.35, P<0.001), suggesting a restrictive filling pattern; the Tei index was significantly increased (0.74±0.15 vs 0.39±0.08, P<0.001), reflecting simultaneous impairment of both overall myocardial systolic and diastolic function. See Table 3 for details. Figure 2 .

[0036] Table 3 Comparison of echocardiographic parameters between the two groups of patients (x̄±s)

[0037] 2.4 Correlation analysis between plasma biochemical indicators and echocardiographic parameters Pearson correlation analysis of plasma biochemical parameters and echocardiographic parameters in the study group showed that: MMP-9 levels were significantly positively correlated with LVEDD (r=0.653), LVESD (r=0.682), E / A ratio (r=0.587), and Tei index (r=0.642) (all P<0.01), and significantly negatively correlated with LVEF (r=-0.715) and LVFS (r=-0.698) (all P<0.01). 5-HT levels were significantly positively correlated with LVEDD (r=0.598), LVESD (r=0.624), E / A ratio (r=0.564), and Tei index (r=0.615) (all P<0.01), and significantly negatively correlated with LVEF (r=-0.683) and LVFS (r=-0.654) (all P<0.01). The NE level was significantly positively correlated with LVEDD (r=0.675), LVESD (r=0.694), E / A ratio (r=0.602), and Tei index (r=0.659) (all P<0.01), and significantly negatively correlated with LVEF (r=-0.728) and LVFS (r=-0.712) (all P<0.01). ACh levels were significantly negatively correlated with LVEDD (r=-0.612), LVESD (r=-0.638), E / A ratio (r=-0.573), and Tei index (r=-0.629) (all P<0.01), and significantly positively correlated with LVEF (r=0.695) and LVFS (r=0.674) (all P<0.01).

[0038] See Table 4 for details. Figure 3 .

[0039] Table 4. Correlation analysis (r-value) between plasma biochemical indicators and echocardiographic parameters.

[0040] 2.5 Determination of Diagnostic Thresholds In summary, the following thresholds were determined for the diagnosis of high-risk ventricular arrhythmias: MMP-9 ≥16.85 ng / mL 5-HT ≥756.32 ng / mL NE ≥85.46 ng / L ACh ≤28.45 ng / mL 2.6 Prognostic Assessment In the study group, elevated levels of MMP-9, 5-HT, and NE, and decreased levels of ACh were significantly associated with worsening cardiac function and an increased risk of ventricular arrhythmias, which could predict patient prognosis. Combining ultrasound parameters could enhance the accuracy of the prediction results.

[0041] 3. Conclusion (1) Precision diagnosis: Combining MMP-9, 5-HT, NE, and ACh can improve the sensitivity and specificity of diagnosis of ventricular arrhythmias in DCM, which is significantly better than that of a single indicator.

[0042] (2) Risk stratification: MMP-9, 5-HT, NE, and ACh levels are significantly correlated with cardiac structural and functional parameters and can help differentiate high-risk patients. If MMP-9 ≥ 16.85 ng / mL, 5-HT ≥ 756.32 ng / mL, NE ≥ 85.46 ng / L, and ACh ≤ 28.45 ng / mL, combined with ultrasound parameters (such as LVEF < 30%), the patient is diagnosed with DCM accompanied by high-risk ventricular arrhythmias.

[0043] (3) Prognostic prediction: High levels of MMP-9, 5-HT, and NE, combined with low ACh levels, low LVEF, and high Tei index, can effectively identify patients with poor prognosis and guide clinical intervention.

[0044] In summary, based on the findings of this invention, measuring plasma levels of MMP-9, 5-HT, NE, and ACh can effectively identify patients with diabetic cerebrovascular disease (DCM) accompanied by ventricular arrhythmias, assess the severity of their condition, and predict prognostic risks. This is of great significance for improving management strategies for DCM patients, optimizing treatment plans, and reducing medical costs.

[0045] The above descriptions are merely embodiments of the present invention, and common knowledge such as specific technical solutions and / or characteristics are not described in detail here. It should be noted that those skilled in the art can make various modifications and improvements without departing from the technical solutions of the present invention, and these should also be considered within the scope of protection of the present invention. These modifications and improvements will not affect the effectiveness of the implementation of the present invention or the practicality of the patent. The scope of protection claimed in this application should be determined by the content of its claims, and the specific embodiments described in the specification can be used to interpret the content of the claims.

Claims

1. Biomarkers for the diagnosis and prognostic assessment of ventricular arrhythmias in dilated cardiomyopathy, characterized by: The biomarkers include a combination of MMP-9, 5-HT, NE, and ACh.

2. Application of products that detect MMP-9, 5-HT, NE and / or ACh in the preparation of tools for diagnosing ventricular arrhythmias in dilated cardiomyopathy or predicting its prognosis.

3. The application according to claim 2, characterized in that: The products used to detect MMP-9, 5-HT, NE, or / and ACh include those that detect the expression levels of MMP-9, 5-HT, NE, or / and ACh.

4. The application according to claim 3, characterized in that: The product was used to measure the levels of MMP-9, 5-HT, NE, and ACh in the subject's sample. Compared with normal individuals, the expression levels of MMP-9, 5-HT, and NE in the subject's sample were increased, while the expression level of ACh was decreased, which suggests that the subject has ventricular arrhythmia or a poor prognosis.

5. The application according to claim 4, characterized in that: The product is used to measure the levels of MMP-9, 5-HT, NE, and ACh in subject samples and compare these levels with predetermined thresholds for the diagnosis and risk stratification of dilated cardiomyopathy with ventricular arrhythmias.

6. The application according to claim 4 or 5, characterized in that: The subject samples were derived from plasma.

7. A tool for diagnosing ventricular arrhythmias in dilated cardiomyopathy or predicting its prognosis, characterized in that: The tool includes a data analysis module and reagents for detecting biomarkers MMP-9, 5-HT, NE, and ACh. The data analysis module is used to analyze the detection values ​​of biomarkers in subject samples.

8. The tool according to claim 7, characterized in that: Compared with normal individuals, when the levels of MMP-9, 5-HT, and / or NE are elevated and the level of ACh is decreased, it is considered that the subject has ventricular arrhythmia. Furthermore, MMP-9, 5-HT, and NE are positively correlated with the degree of cardiac structural and functional damage, while ACh is negatively correlated with the degree of cardiac structural and functional damage. When the MMP-9 level is ≥16.85 ng / mL, 5-HT is ≥756.32 ng / mL, NE is ≥85.46 ng / L, and ACh is ≤28.45 ng / mL, the subject is considered to have dilated cardiomyopathy with high-risk ventricular arrhythmia.

9. The tool according to claim 8, characterized in that: The biomarkers also include echocardiographic parameters, including LVEDD, LVESD, LVEF, LVFS, E / A ratio, and Tei index, which are used in conjunction with MMP-9, 5-HT, NE, and ACh to assess the risk of ventricular arrhythmias and cardiac function status.

10. The tool according to claim 7, characterized in that: The subject samples were derived from plasma.

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