Application of NPRA as target spot in treatment or prevention of diabetic cardiomyopathy

Abstract

The invention discloses application of NPRA as a target spot in treatment or prevention of diabetic cardiomyopathy, further discloses application of NPRA as a target spot in drugs for treatment or prevention of diabetic cardiomyopathy, and solves the problems that the pathogenesis of diabetic cardiomyopathy is unclear, the diabetic cardiomyopathy is not easy to detect in the early stage, and the diabetic cardiomyopathy is difficult to treat or prevent.

Description

Technical Field

[0001] The present invention belongs to the field of medical technology, and specifically relates to the application of NPRA as a target in treating or preventing diabetic cardiomyopathy. Background Art

[0002] Diabetes is one of the most common chronic diseases in the world. Diabetic cardiomyopathy is one of the most serious and common complications of diabetes. Its pathogenesis is not yet clear, and it is difficult to detect in the early stage. The lesions can easily progress to end-stage heart failure, which can eventually lead to more than 75% of diabetic patients dying from heart failure. Therefore, exploring the pathogenesis and conducting early intervention are of great significance for the prevention and treatment of diabetic cardiomyopathy.

[0003] The occurrence of cardiomyopathy is related to the reduced effect of endogenous protective peptides in the myocardium. Previous studies have found that the level of natriuretic peptides in plasma can be used as one of the diagnostic indicators of diabetic cardiomyopathy. Natriuretic peptides are a group of polypeptides with similar structures and functions. They mainly play the functions of diuresis, sodium excretion, maintaining the body's water and salt balance, and vascular homeostasis. They mainly exert their effects by binding to receptors.

[0004] In cardiovascular diseases, natriuretic peptide receptor A (NPRA) mainly plays a cardioprotective role. Currently, the molecular and cellular functions of NPRA are not very clear. Studies have found that the expression of NPRA in the myocardial tissue of diabetic mice is decreased, but there is no report to confirm whether NPRA is involved in and how it regulates the occurrence and development of diabetic cardiomyopathy. Summary of the invention

[0005] The first objective of the present invention is to provide the use of NPRA as a target in the treatment or prevention of diabetic cardiomyopathy, which solves the problem that diabetic cardiomyopathy is difficult to detect in the early stage and difficult to treat or prevent due to the unclear pathogenesis.

[0006] The second object of the present invention is to provide the use of NPRA as a target in drugs for treating or preventing diabetic cardiomyopathy.

[0007] The first technical solution adopted by the present invention is: using NPRA as a target in the treatment or prevention of diabetic cardiomyopathy.

[0008] The first technical solution adopted by the present invention is also characterized in that: Furthermore, NPRA is used as a target to prevent diabetes by detecting changes in NPRA in the subjects' plasma, and is related to cardiac function.

[0009] Furthermore, NPRA was used as a target for the treatment of diabetes by injecting adeno-associated virus into the myocardium to overexpress NPRA.

[0010] Furthermore, after overexpression of NPRA, the protein level and mRNA content of NPRA in diabetic myocardial tissue increased significantly, significantly improving cardiac function and inhibiting myocardial hypertrophy.

[0011] The second technical solution adopted by the present invention is: the use of NPRA as a target in drugs for treating or preventing diabetic cardiomyopathy.

[0012] The beneficial effects of the present invention are: The present invention provides the use of NPRA as a target in the treatment or prevention of diabetic cardiomyopathy, which solves the problem that diabetic cardiomyopathy is difficult to detect in the early stage and difficult to treat or prevent due to the unclear pathogenesis.

[0013] The present invention provides a method for significantly increasing the protein level and mRNA content of NPRA in diabetic myocardial tissue, significantly improving cardiac function, and inhibiting myocardial hypertrophy by injecting adeno-associated virus overexpressing NPRA (AAV9-NPRA) at myocardial points. BRIEF DESCRIPTION OF THE DRAWINGS

[0014] Figure 1 It is a graph of human heart left ventricular ejection fraction and plasma NPRA content; Figure 2 is a graph of left ventricular ejection fraction and plasma NPRA content in mice; Figure 3 This is a graph of NPRA protein and mRNA expression in mouse myocardium; Figure 4 This is the expression diagram of NPRA protein and mRNA in myocardial tissue of myocardial-specific knockout Npr1 mice; Figure 5 This is a graph showing left ventricular ejection fraction, myocardial cross-sectional area, and heart-to-weight ratio in the heart of myocardium-specific knockout Npr1 mice; Figure 6 This is the expression diagram of NPRA protein and mRNA in myocardial tissue of mice overexpressing Npr1 after the application of adeno-associated virus (AAV); Figure 7 This is a graph of left ventricular ejection fraction, myocardial cross-sectional area and heart-to-weight ratio in mice overexpressing Npr1 in the myocardium after the application of adeno-associated virus (AAV). DETAILED DESCRIPTION

[0015] The technical solution of the present invention will be clearly and completely described below in conjunction with the accompanying drawings.

[0016] The present invention provides the use of NPRA as a target in the treatment or prevention of diabetic cardiomyopathy. The prevention of diabetes using NPRA as a target is implemented by detecting the changes in NPRA in the plasma of a subject, and is positively correlated with cardiac function. The treatment of diabetes using NPRA as a target is implemented by injecting adeno-associated virus into the myocardium to overexpress NPRA. After overexpressing NPRA, the protein level and mRNA content of NPRA in diabetic myocardial tissue are significantly increased, cardiac function is significantly improved, and myocardial hypertrophy is inhibited.

[0017] The present invention also provides the use of NPRA as a target in drugs for treating or preventing diabetic cardiomyopathy. The relationship between NPRA expression and diabetic cardiomyopathy is further determined below through examples and experimental figures: Example 1 The level of NPRA in plasma is significantly decreased in patients with diabetic myocardial injury and is positively correlated with cardiac function.

[0018] The present invention uses ELISA technology to detect the content of NPRA in the plasma of diabetic patients and patients with diabetic myocardial injury. The results show that compared with diabetic patients, the content of NPRA in the plasma of patients with diabetic myocardial injury is significantly reduced; through further correlation analysis, it is found that the content of NPRA in the plasma of patients with diabetic myocardial injury is positively correlated with cardiac function, indicating that NPRA is closely related to the occurrence and development of diabetic cardiomyopathy. Figure 1 As shown, Figure 1 The left side shows the left ventricular ejection fraction detected by cardiac ultrasound. Figure 1 The right side shows the plasma NPRA content, n=6.

[0019] Example 2 The NPRA content in mouse plasma was significantly decreased in patients with high-fat-induced diabetic myocardial injury and was positively correlated with cardiac function.

[0020] In the present invention, wild-type C57BL / 6 mice (WT) were used as experimental subjects, and a mouse diabetic cardiomyopathy (DCM) model was established by inducing a small amount of STZ combined with a high-fat diet. The content of NPRA in plasma was detected by ELISA experiments. The results showed that compared with WT mice, the content of NPRA in the plasma of DCM mice was significantly reduced and positively correlated with cardiac function. The above results indicate that the changes in NPRA in the myocardium of DCM mice are consistent with those in patients with diabetic cardiomyopathy, such as Figure 2 As shown, Figure 2 Left ventricular ejection fraction measured by left heart ultrasound, Figure 2 The right side shows the plasma NPRA content, n=6.

[0021] Example 3 The expression level of NPRA in mouse myocardial tissue was significantly downregulated in the high-fat-induced diabetic cardiomyopathy model.

[0022] This study used wild-type C57BL / 6 mice (WT) as experimental subjects, established a mouse diabetic cardiomyopathy (DCM) model by inducing a small amount of STZ combined with a high-fat diet, and detected the protein and mRNA expression changes of NPRA in the mouse heart tissue by Western blotting (WB) and real-time quantitative PCR (RT-PCR) experimental methods. The results showed that compared with WT mice, the protein expression level and mRNA expression content of NPRA in the heart tissue of DCM mice were significantly downregulated, indicating that NPRA is likely to be involved in the occurrence and development of diabetic cardiomyopathy. Figure 3 As shown, Figure 3 The left side shows the protein expression of NPRA in mouse myocardial tissue. Figure 3 The middle one shows the statistical graph of NPRA protein expression in mouse myocardial tissue. Figure 3 The right side shows the relative expression level of NPRA mRNA in mouse myocardial tissue, n=6.

[0023] Example 4 Cardiac-specific knockout of npr1 exacerbates high-fat diet-induced cardiac hypertrophy and deterioration of cardiac function in mice.

[0024] The present invention uses cardiac-specific npr1 gene knockout mice (npr1- / -) as the experimental group and the flox group of cardiac-specific npr1 gene knockout mice (npr1flox / flox) as the control group. A mouse diabetic cardiomyopathy model is constructed by inducing a small amount of STZ combined with high-fat feeding to study the role of the npr1 gene in diabetic cardiomyopathy. The results showed that compared with the control group npr1flox / flox mice, the npr1flox / flox mice significantly aggravated myocardial hypertrophy and deterioration of cardiac function caused by diabetic myocardium. This indicates that the inhibition of npr1 exacerbates the occurrence and development of diabetic cardiomyopathy, such as Figure 4 As shown, Figure 4 The left side shows the protein expression of NPRA in mouse myocardial tissue. Figure 4 The middle one shows the statistical graph of NPRA protein expression in mouse myocardial tissue. Figure 4 The right side shows the relative expression level of NPRA mRNA in mouse myocardial tissue, n=6.

[0025] Example 5 Overexpression of the npr1 gene in the myocardium by adeno-associated virus alleviates high-fat diet-induced myocardial hypertrophy and deterioration of cardiac function in mice.

[0026] The present invention established a mouse diabetic cardiomyopathy (DCM) model by inducing a small amount of STZ combined with a high-fat diet, and injected adeno-associated virus to overexpress NPRA (AAV9-NPRA) through myocardial point injection to compare the changes in myocardial NPRA and cardiac function in diabetic cardiomyopathy mice after overexpression of NPRA. The results showed that after overexpression of NPRA, the protein level and mRNA content of NPRA in diabetic myocardial tissue increased significantly, significantly improving cardiac function. Overexpression of NPRA may be used as an intervention target for the prevention and treatment of diabetic cardiomyopathy, such as Figure 5 As shown, Figure 5 The left side indicates the left ventricular ejection fraction of the mouse heart. Figure 5 The middle represents the cross-sectional area of ​​mouse cardiomyocytes. Figure 5 The right side shows the ratio of heart weight to tibia length in mouse heart tissue, n=6.

[0027] Example 6 Overexpression of Npr1 gene in myocardium by adeno-associated virus significantly increased the protein expression and mRNA content of NPRA.

[0028] In this study, a mouse model of diabetic cardiomyopathy (DCM) was established by inducing a small amount of STZ combined with a high-fat diet. Adeno-associated virus (AAV9-NPRA) was injected into the myocardium to overexpress NPRA. The expression of NPRA in the myocardium of diabetic cardiomyopathy mice was compared after NPRA was overexpressed. The results showed that the protein level and mRNA content of NPRA in diabetic myocardial tissue increased significantly after NPRA was overexpressed. Figure 6 As shown, Figure 6 The left side shows the protein expression of NPRA in mouse myocardial tissue. Figure 6 The middle one shows the statistical graph of NPRA protein expression in mouse myocardial tissue. Figure 6 The right side shows the relative expression level of NPRA mRNA in mouse myocardial tissue, n=6.

[0029] Example 7 Overexpression of Npr1 gene in myocardium by adeno-associated virus alleviates high-fat diet-induced myocardial hypertrophy and deterioration of cardiac function in mice.

[0030] In this study, a mouse model of diabetic cardiomyopathy (DCM) was established by inducing a small amount of STZ combined with a high-fat diet. Adeno-associated virus overexpressed NPRA (AAV9-NPRA) was injected into the myocardium to compare the changes in cardiac function and structure in mice with diabetic cardiomyopathy after overexpression of NPRA. The results showed that overexpression of NPRA in the myocardium significantly improved the cardiac function and structure of mice. This suggests that overexpression of NPRA may be an intervention target for the prevention and treatment of diabetic cardiomyopathy. Figure 7 As shown, Figure 7 The left side shows the left ventricular ejection fraction detected by cardiac ultrasound. Figure 7The middle is the cross-sectional area of ​​the myocardium. Figure 7 The right side shows the ratio of heart weight to tibia length, n=6.

Claims

1. Application of NPRA as a target in the treatment or prevention of diabetic cardiomyopathy.

2. The use of NPRA as a target in the treatment or prevention of diabetic cardiomyopathy according to claim 1, characterized in that: The NPRA is used as a target to prevent diabetes by detecting changes in NPRA in the plasma of subjects, and is positively correlated with cardiac function.

3. Use of NPRA as a target in the treatment or prevention of diabetic cardiomyopathy according to claim 1 or 2, characterized in that: The NPRA is used as a target to treat diabetes by injecting adeno-associated virus into the myocardium to overexpress NPRA.

4. The use of NPRA as a target in the treatment or prevention of diabetic cardiomyopathy according to claim 3, characterized in that: After overexpression of NPRA, the protein level and mRNA content of NPRA in diabetic myocardial tissue increased significantly, significantly improved cardiac function, and inhibited myocardial hypertrophy.

5. Application of NPRA as a target in drugs for the treatment or prevention of diabetic cardiomyopathy.

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