Biomarker of Parkinson's disease and application thereof

Abstract

The invention relates to the technical field of biological medicine, in particular to a Parkinson's disease biomarker and application thereof. The application of the NTN1 protein, the RELN protein and / or the SLIT1 protein or the transcription product mRNA or the mRNA fragment thereof as the biomarker in any one of the following items: 1) application in preparation of products for diagnosis or auxiliary diagnosis of Parkinson's disease of human or non-human mammals, and 2) application in preparation of products for diagnosis or auxiliary diagnosis of Parkinson's disease of human or non-human mammals; 2) application in preparation of products for screening or auxiliary screening of Parkinson's disease of human or non-human mammals; 3) application in preparation of products for diagnosis or auxiliary diagnosis of heart failure caused by Parkinson's disease of human or non-human mammals; 4) application in preparation of products for screening or auxiliary screening of heart failure caused by Parkinson's disease of human or non-human mammals; the invention discloses a PD-related hub gene behind heart failure and a potential mechanism, and determines that biomarkers (RELN, SLIT1 and NTN1) can be used as biomarkers for the risk of heart failure caused by PD.

Description

Technical Field

[0001] The present invention relates to the field of biomedical technology, and in particular to a Parkinson's disease biomarker and a use thereof. Background Art

[0002] Parkinson's disease (PD) is a global progressive neurodegenerative disease that currently affects 1% to 3% of people over the age of 65. An increasing number of studies have shown that PD is manifested not only by a decline in brain function, but also by inflammatory cascades and oxidative stress, which are closely related to the pathogenesis of various cardiovascular diseases, including myocardial infarction, ischemic stroke, and heart failure. Heart failure (HF) affects more than 60 million people worldwide, with an estimated current prevalence of 2% in the adult population, and is the most common cardiovascular cause of hospitalization in patients over the age of 60. Recent studies have shown that the frequency of observation of HF in PD patients is higher than that in the general population, indicating that PD is associated with an increased risk of HF development. Cardiovascular changes such as orthostatic hypotension, heart rate variability, changes in ECG parameters, and baroreflex dysfunction can occur in the early and late stages of PD and worsen with the progression of the disease. Cardiovascular changes may affect about 80% of PD patients and may exacerbate the progression of the disease and increase the risk of death.

[0003] Recent evidence has shown a strong link between Parkinson's disease (PD) and cardiac dysfunction, but treatment options are limited. Studies investigating the association between genetic or environmental factors and PD have revealed many factors shared with heart failure (HF), such as older age, male sex, and diabetes. In addition, common inflammatory pathways, glucose metabolism, oxidative stress, lipid metabolism, and shared risk factors are thought to play a role in this process. PD-associated genes are also expressed in the heart, such as Parkin, PINK1, DJ-1, and LRRK2, and some studies have shown that the above proteins can protect the heart from damage. Noncoding RNAs also play a role in regulating common pathways in PD and heart disease, such as miR-124, miR-133b, lncRNA MALAT1, and HOTAIR. These studies suggest that PD may lead to subsequent complications, including heart failure (HF), at least in part through secreted proteins. There is currently no effective biomarker for diagnosing the likelihood of HF in patients with PD, so there is a need to provide an effective biomarker for the clinical diagnosis of PD and HF. Summary of the invention

[0004] Therefore, the technical problem to be solved by the present invention is to provide a Parkinson's biomarker and its use.

[0005] To this end, the present invention provides the following technical solutions:

[0006] The embodiments of the present invention disclose the use of NTN1 protein, RELN protein and / or SLIT1 protein or their transcription product mRNA or mRNA fragment as a biomarker in any of the following:

[0007] 1) Application in the preparation of products for diagnosing or assisting in the diagnosis of Parkinson's disease in humans or non-human mammals;

[0008] 2) Application in the preparation of products for screening or assisting screening of Parkinson's disease in humans or non-human mammals;

[0009] 3) Application in the preparation of products for diagnosing or assisting in the diagnosis of heart failure caused by Parkinson's disease in humans or non-human mammals;

[0010] 4) Application in the preparation of products for screening or assisting screening of heart failure caused by Parkinson's disease in humans or non-human mammals.

[0011] The embodiments of the present invention disclose a product for detecting NTN1 protein, RELN protein and / or SLIT1 protein or their transcription product mRNA or mRNA fragments for any of the following purposes:

[0012] 1) Application in the preparation of products for diagnosing or assisting in the diagnosis of Parkinson's disease in humans or non-human mammals;

[0013] 2) Application in the preparation of products for screening or assisting screening of Parkinson's disease in humans or non-human mammals;

[0014] 3) Application in the preparation of products for diagnosing or assisting in the diagnosis of heart failure caused by Parkinson's disease in humans or non-human mammals;

[0015] 4) Application in the preparation of products for screening or assisting screening of heart failure caused by Parkinson's disease in humans or non-human mammals.

[0016] In some embodiments, the product for detecting NTN1 protein, RELN protein and / or SLIT1 protein or their transcription products mRNA or mRNA fragments includes a product for detecting the expression level of NTN1 protein, RELN protein and / or SLIT1 protein or their transcription products mRNA or mRNA fragments in the test sample.

[0017] In some embodiments, the sample to be tested is cardiac tissue or blood of a human or non-human mammal. In a preferred embodiment, the sample to be tested is selected from blood.

[0018] In some embodiments, the product for detecting NTN1 protein, RELN protein and / or SLIT1 protein or their transcription products mRNA or mRNA fragments includes a kit, a reagent, a test strip, a primer, a probe or a chip.

[0019] In some embodiments, the reagent includes: an antibody, protein and / or polypeptide that specifically binds to NTN1 protein, RELN protein and / or SLIT1 protein; or a reagent that detects mRNA or mRNA fragments of NTN1 protein, RELN protein and / or SLIT1 protein transcription products;

[0020] In some embodiments, the antibody, protein and / or polypeptide is modified with an isotope, horseradish peroxidase, colloidal gold or a fluorescent probe, or is unmodified.

[0021] In some embodiments, the reagent for detecting the mRNA or mRNA fragment of the transcription product of NTN1 protein, RELN protein and / or SLIT1 protein includes a PCR reaction buffer or a nucleic acid molecule hybridization buffer; optionally, it includes gene fragments of NTN1 protein, RELN protein and / or SLIT1 protein or specific sequence fragments of their mRNA, or molecular probes containing these specific sequence fragments.

[0022] The embodiments of the present invention disclose a product for diagnosing, assisting in the diagnosis, screening or assisting in the screening of Parkinson's disease or Parkinson's disease-induced heart failure in humans or non-human mammals, including a product for detecting the expression level of NTN1 protein, RELN protein and / or SLIT1 protein or their transcription product mRNA or mRNA fragment in a sample to be tested.

[0023] In some embodiments, the product for detecting NTN1 protein, RELN protein and / or SLIT1 protein or their transcription products mRNA or mRNA fragments includes a product for detecting the expression level of NTN1 protein, RELN protein and / or SLIT1 protein or their transcription products mRNA or mRNA fragments in the test sample.

[0024] In some embodiments, the sample to be tested is cardiac tissue or blood of a human or non-human mammal. In a preferred embodiment, the sample to be tested is selected from blood.

[0025] In some embodiments, the product for detecting NTN1 protein, RELN protein and / or SLIT1 protein or their transcription products mRNA or mRNA fragments includes a kit, a reagent, a test strip, a primer, a probe or a chip.

[0026] In some embodiments, the reagent includes: an antibody, protein and / or polypeptide that specifically binds to NTN1 protein, RELN protein and / or SLIT1 protein; or a reagent that detects mRNA or mRNA fragments of NTN1 protein, RELN protein and / or SLIT1 protein transcription products;

[0027] In some embodiments, the antibody, protein and / or polypeptide is modified with an isotope, horseradish peroxidase, colloidal gold or a fluorescent probe, or is unmodified.

[0028] In some embodiments, the reagent for detecting the mRNA or mRNA fragment of the transcription product of NTN1 protein, RELN protein and / or SLIT1 protein includes a PCR reaction buffer or a nucleic acid molecule hybridization buffer; optionally, it includes gene fragments of NTN1 protein, RELN protein and / or SLIT1 protein or specific sequence fragments of their mRNA, or molecular probes containing these specific sequence fragments.

[0029] In some embodiments, the non-human mammal comprises an animal model of Parkinson's disease, an animal model of Parkinson's disease-induced heart failure, or an animal model induced by MPTP.

[0030] In some embodiments, the human is a patient with Parkinson's disease, or a patient with Parkinson's disease-induced heart failure.

[0031] The present invention discloses a device for diagnosing, assisting in diagnosis, screening or assisting in screening Parkinson's disease or Parkinson's disease-induced heart failure in humans or non-human mammals, comprising:

[0032] A detection unit, used to obtain the expression level of NTN1 protein, RELN protein and / or SLIT1 protein or their transcription product mRNA or mRNA fragment in the sample to be tested;

[0033] The judgment unit is used to diagnose or assist in diagnosis, screening or assist in screening whether a human or non-human mammal suffers from Parkinson's disease or Parkinson's-induced heart failure based on the expression level of NTN1 protein, RELN protein and / or SLIT1 protein or their transcription product mRNA or mRNA fragment in the test sample.

[0034] In some embodiments, the expression level of NTN1 protein, RELN protein and / or SLIT1 protein or their transcription product mRNA or mRNA fragments in the sample of the human or non-human mammal to be tested is reduced compared with the expression level of NTN1 protein, RELN protein and / or SLIT1 protein or their transcription product mRNA or mRNA fragments in the sample of a normal human or normal non-human mammal, and the human or non-human mammal to be tested is judged to suffer from Parkinson's disease or Parkinson's-induced heart failure.

[0035] The present invention discloses a method for screening drugs for treating Parkinson's disease or Parkinson's disease-induced heart failure, comprising:

[0036] The candidate drugs are used in the non-human mammal model of Parkinson's disease or Parkinson's disease induced heart failure; the drugs that up-regulate the expression of NTN1 protein, RELN protein and / or SLIT1 protein or their transcription product mRNA or mRNA fragments or enhance their activity are screened, or the candidate drugs that alleviate Parkinson's disease or Parkinson's disease induced heart failure are screened as the drugs for treating Parkinson's disease or Parkinson's disease induced heart failure.

[0037] The technical solution of the present invention has the following advantages:

[0038] 1. Use of the NTN1 protein, RELN protein and / or SLIT1 protein or their transcription product mRNA or mRNA fragments provided by the present invention as biomarkers in any of the following items: 1) in the preparation of products for diagnosing or assisting in the diagnosis of Parkinson's disease in humans or non-human mammals; 2) in the preparation of products for screening or assisting in the screening of Parkinson's disease in humans or non-human mammals; 3) in the preparation of products for diagnosing or assisting in the diagnosis of heart failure caused by Parkinson's disease in humans or non-human mammals; 4) in the preparation of products for screening or assisting in the screening of heart failure caused by Parkinson's disease in humans or non-human mammals; The present invention reveals the hub genes and potential mechanisms behind PD-related heart failure, and identifies three biomarkers (RELN, SLIT1 and NTN1) through analysis and verification. The proteins encoded by these three genes can be detected in the blood and can be used as biomarkers for the risk of heart failure in PD.

[0039] Furthermore, the present invention validated one of the genes, NTN1, in PD model mice, and the results showed that NTN1 expression in the hippocampus, blood, and heart of PD model mice changed, and the NTN1 content in the heart was negatively correlated with fibrosis. These findings provide a new perspective on the coexistence of PD and HF, especially focusing on preventing heart failure in PD patients and using these mechanisms as a means of cardioprotection.

[0040] Furthermore, the sample to be tested in the present invention can be blood, and the test is convenient and easy to operate.

[0041] Furthermore, NTN1, RELN and / or SLIT1 can serve as targets for the simultaneous treatment of Parkinson's disease and heart failure. BRIEF DESCRIPTION OF THE DRAWINGS

[0042] In order to more clearly illustrate the specific implementation methods of the present invention or the technical solutions in the prior art, the drawings required for use in the specific implementation methods or the description of the prior art will be briefly introduced below. Obviously, the drawings described below are some implementation methods of the present invention. For ordinary technicians in this field, other drawings can be obtained based on these drawings without paying creative work.

[0043] Figure 1 is a principal component analysis cluster diagram of the merger of three Parkinson's gene data sets in Example 1 of the present invention;

[0044] Figure 2 is the differentially expressed secretory protein in PD screened in Example 1 of the present invention;

[0045] Figure 3 It is the potential functional gene for screening Parkinson's disease-induced heart failure in Example 1 of the present invention;

[0046] Figure 4 It is the key gene identified by PPI network analysis in Example 1 of the present invention;

[0047] Figure 5 This is the rotarod test of each group of mice in Example 2 of the present invention;

[0048] Figure 6 is a cardiac pathology diagram of each group of mice in Example 2 of the present invention;

[0049] Figure 7 is the correlation between NTN1 and cardiac fibrosis ratio in the hearts of control group mice and model group mice in Example 2 of the present invention;

[0050] Figure 8 is the content of NTN1 protein (Netrin-1) in the blood of the model and control groups;

[0051] Fig. 9 is the RELN protein (Reelin) content in the blood of the model and control groups;

[0052] Fig.10 is the expression level of NTN1 in the heart failure validation dataset GSE1145;

[0053] Fig.11 is the expression level of RELN in the heart failure validation dataset GSE1145;

[0054] Fig.12 is the ROC curve for predicting heart failure using NTN1 and RELN;

[0055] Fig.13 This is the ROC curve for predicting heart failure using NTN1;

[0056] Fig.14 This is the ROC curve for predicting heart failure using RELN. DETAILED DESCRIPTION

[0057] The following examples are provided for a better understanding of the present invention, but are not intended to limit the best mode of implementation, nor to limit the content and protection scope of the present invention. Any product identical or similar to the present invention obtained by anyone under the inspiration of the present invention or by combining the features of the present invention with other prior arts shall fall within the protection scope of the present invention.

[0058] If no specific experimental steps or conditions are specified in the examples, the conventional experimental steps or conditions described in the literature in the field can be used. If no manufacturer is specified for the reagents or instruments used, they are all conventional reagent products that can be obtained commercially.

[0059] Example 1 Screening of markers

[0060] (1) Sample acquisition

[0061] The PD datasets GSE7621, GSE20163, and GSE49036 were downloaded from the Gene Expression Omnibus (GEO, https: / / www.ncbi.nlm.nih.gov / geo / ) and batch corrected to obtain comprehensive PD expression data: including expression data of 27 healthy controls (HC) and 41 PD patients. The principal component analysis (PCA) clustering diagram was used to visualize the effect of removing batch differences. The results are shown in Figure 2. Figure 1 shown.

[0062] (2) Identification of differentially expressed genes (DEGs) between Parkinson's disease (PD) and normal samples

[0063] The differential analysis of the expressed genes in 27 healthy controls (HC) and 41 PD patients was performed using the weighted least squares method based on the R language package 'limma' to estimate the differences in gene expression, and the Bayesian method was used to correct for multiple testing problems. 168 differentially expressed genes (DEGs) were revealed, and the screening criteria were corrected p value ≤ 0.05 and log2 (fold change) ≥ 0.5. The final screening results included 52 up-regulated genes and 116 down-regulated genes.

[0064] (3) Identification of differentially expressed secretory proteins in Parkinson's disease (PD)

[0065] Considering that PD may promote the onset and development of heart failure (HF) by releasing secretory proteins, this example then obtained the secretory proteins associated with PD through the intersection of DEGs and secretory proteins in plasma. A total of 21 secretory proteins in plasma related to PD were obtained. Comparative analysis showed that PLBD2, LPO, PCYOX1L, RSPO2, SDC1, RBM3, PBX1, CDH8, NTN1, SUSD1, SEMA3G, ANK1, SLIT1, GNTN4, NTN4, LRRC3B, OLFM3, DLK1, and KIAA0319 were expressed at a lower level in PD, while AKR1C1 and RELN were expressed at a higher level ( Figure 2 ).

[0066] (4) Identification of differentially expressed genes in heart failure (HF)

[0067] In the GSE57338 dataset, a total of 8884 differentially expressed genes (DEGs) between HF and normal samples were identified by setting the corrected p-value ≤ 0.01, including 4171 upregulated genes and 4713 downregulated genes.

[0068] (5) Identification of potential functional genes in Parkinson's disease-induced heart failure

[0069] Since the common differentially expressed secreted proteins between heart failure and Parkinson's disease may play a key role in Parkinson's disease-associated heart failure patients, this study identified 12 common genes at the intersection of Parkinson's disease-associated secreted proteins and heart failure DEGs, as well as the expression of these 12 genes in heart failure samples ( Figure 3 ).

[0070] (6) Identification of key genes through PPI network analysis

[0071] To identify hub genes among the 12 potential functional genes, a protein-protein interaction (PPI) network was constructed using the STRING website and analyzed using Cytoscape, resulting in a subnetwork containing three candidate genes, RELN, SLIT1, and NTN1 ( Figure 4 ).

[0072] Example 2 Validation of NTN1 in a Parkinson's mouse model

[0073] (1) Establishment of Parkinson's mouse model:

[0074] Male C57BL / 6J mice aged 16 months were used. The mice were housed in a 12-hour day-night cycle and had free access to food and water. The mice received intraperitoneal injections of 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) (20 mg / kg) twice a week for 4 weeks, and the control group of animals received an equal volume of 0.9% saline. There were 8 mice in each group.

[0075] (2) Behavioral test:

[0076] At the end of the establishment of the Parkinson's mouse model, all mice received 3 consecutive days of behavioral training before testing. The rotating rod test was performed using an accelerating rotating rod apparatus to assess the motor coordination ability of PD mice. To standardize the challenge, the rotation speed of the test was consistently set at 40 revolutions per minute. The experiment was performed three times. The results are shown in Figure 5 As shown, there was a statistically significant difference between the model (MPTP) group and the control group (p < 0.05), indicating that the constructed Parkinson's mouse model was successfully established.

[0077] (3) Histological analysis:

[0078] After the behavioral test, mice in each group were killed, and heart tissue was obtained. The heart tissue was fixed in 4% formaldehyde and embedded in paraffin. The sections were stained with Masson trichrome to evaluate myocardial fibrosis. The fibrosis area was measured using ImageJ software. The cardiac pathology of mice in each group is shown in Figure 2. Figure 6 As shown by Figure 6 It can be concluded that fibrosis occurs in the heart of the Parkinson's mouse model, and NTN1 expression is reduced, indicating abnormal cardiac function.

[0079] Detection method of NTN1 in cardiac tissue:

[0080] Mouse heart tissue was fixed in 4% paraformaldehyde / 30% sucrose in PBS and then permeabilized with PBS-T (50mM Tris·HCl, 150mM NaCl, 3% bovine serum albumin [BSA], 0.1% Triton-X100, pH 7.4). Sections of the fixed and sectioned samples were incubated with primary antibody Netrin-1, and then sections were incubated with fluorescently labeled secondary antibodies for 1 hour at room temperature. After rinsing three times with PBS, the slides were mounted with glass coverslips using mounting fluid and finally photographed under a fluorescence microscope.

[0081] The correlation between NTN1 in cardiac tissue and the fibrosis area in cardiac tissue was investigated. Figure 7 As shown, it can be concluded that the expression level of NTN1 in the heart is inversely proportional to the proportion of cardiac fibrosis.

[0082] Method for detecting NTN1 protein (Netrin-1) and RELN protein (Reelin) in blood:

[0083] 20ul mouse plasma was added to 20ul RIPA lysis buffer (Biyuntian Catalog No. P0013B) and lysed at 4°C for 1h. The fully automated protein blotting system and matching kit (Simple Western, Catalog No. SM-W004) were used. NTN1 antibody was purchased from Immunoway (Catalog No. YT3042), and RELN protein was purchased from Huaan Biotechnology (Catalog No. ET1704-98). The sample volume per well was 2ul, and the operation was performed according to the instructions.

[0084] The results are as follows Figure 8 and Fig. 9 The results show that the protein levels of NTN1 and RELN in the blood of Parkinson's model mice and control group are compared. Netrin-1 is reduced in the blood of Parkinson's model mice, while Reelin is increased in the blood of Parkinson's model mice. The results show that the protein expression of Netrin-1 and Reelin in Parkinson's mouse model has the same expression changes as Parkinson's patients. These two proteins can be used as biomarkers to predict Parkinson's.

[0085] Example 3

[0086] (1) Establishment of Parkinson's mouse model:

[0087] Male C57BL / 6J mice aged 16 months were used. The mice were housed in a 12-hour day-night cycle environment and had free access to food and water. The mice received intraperitoneal injections of 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) (20 mg / kg) twice a week for 4 weeks, and the control group of animals received an equal amount of 0.9% saline. There were 20 mice in each group.

[0088] (2) Behavioral test:

[0089] At the end of the establishment of the Parkinson's mouse model, all mice received 3 consecutive days of behavioral training before testing. The rotating rod test was performed using an accelerating rotating rod apparatus to assess the motor coordination ability of PD mice. In order to standardize the challenge, the rotation speed of the test was consistently set to 40 revolutions per minute. The experiment was performed three times. The results showed that there was a statistically significant difference between the model (MPTP) group and the control group (p < 0.05), indicating that the constructed Parkinson's mouse model was successfully constructed.

[0090] (3) Histological analysis and NTN1 protein detection in blood

[0091] The histological analysis was the same as in Example 2, and the analysis results were consistent with those of the mice in each group in Example 2.

[0092] Method for detecting NTN1 protein (Netrin-1) and RELN protein (Reelin) in blood:

[0093] 20ul mouse plasma was added to 20ul RIPA lysis buffer (Biyuntian Catalog No. P0013B) and lysed at 4°C for 1h. The fully automated protein blotting system and matching kit (Simple Western, Catalog No. SM-W004) were used. NTN1 antibody was purchased from Immunoway (Catalog No. YT3042), and RELN protein was purchased from Huaan Biotechnology (Catalog No. ET1704-98). The sample volume per well was 2ul, and the operation was performed according to the instructions.

[0094] The results showed that the content of NTN1 and RELN proteins in the blood of Parkinson's model and control group was compared. Compared with the control group, the content of Netrin-1 in the blood of Parkinson's model mice was significantly reduced (p < 0.05), while Reelin was increased in the blood of Parkinson's model mice (p < 0.001). The results show that the protein expression of Netrin-1 and Reelin in the Parkinson's mouse model has the same expression changes as that of Parkinson's patients. These two proteins can be used to predict Parkinson's biomarkers. The above shows that using NTN1 protein or RELN protein or its transcription product mRNA or mRNA fragment as a marker of Parkinson's-induced heart failure has a very accurate predictive ability and can be used to predict whether the mouse animal model of Parkinson's-induced heart failure is successfully constructed. At the same time, it can also objectively diagnose patients with Parkinson's-induced heart failure.

[0095] Example 4

[0096] The heart failure dataset GSE1145 was downloaded from the Gene Expression Omnibus (GEO, https: / / www.ncbi.nlm.nih.gov / geo / ), which includes 10 healthy controls and 24 heart failure patients. The expression levels of NTN1 and RELN proteins in each group were counted. The results are as follows: Fig.10 and Fig.11 As shown in Figure 2, NTN1 expression is decreased in patients with heart failure, while RELN expression is increased in patients with heart failure. The expression levels of NTN1 and RELN are used to judge heart failure patients, such as Fig.12 As shown in Figure 2, the area under the ROC curve obtained an AUC value of 0.883, indicating that the combined prediction of heart failure by NTN1 and RELN has a high accuracy. Fig.13 As shown in Figure 2, the area under the ROC curve obtained an AUC value of 0.69, indicating that NTN1 alone has a certain predictive performance in predicting heart failure. The expression of RELN alone was used to judge heart failure patients, such as Fig.14As shown in the figure, the area under the ROC curve obtained an AUC value of 0.863, indicating that RELN alone has a high accuracy in predicting heart failure.

[0097] The above shows that using NTN1 protein or RELN protein or their transcription product mRNA or mRNA fragment as a marker for Parkinson's disease-induced heart failure has a very accurate predictive ability and can be used to predict patients with Parkinson's disease-induced heart failure.

[0098] Obviously, the above embodiments are merely examples for the purpose of clear explanation, and are not intended to limit the implementation methods. For those skilled in the art, other different forms of changes or modifications can be made based on the above description. It is not necessary and impossible to list all the implementation methods here. The obvious changes or modifications derived therefrom are still within the scope of protection of the invention.

Claims

1. Use of NTN1 protein, RELN protein and / or SLIT1 protein or their transcription product mRNA or mRNA fragment as a biomarker in any of the following: 1) Application in the preparation of products for diagnosing or assisting in the diagnosis of Parkinson's disease in humans or non-human mammals; 2) Application in the preparation of products for screening or assisting screening of Parkinson's disease in humans or non-human mammals; 3) Application in the preparation of products for diagnosing or assisting in the diagnosis of heart failure caused by Parkinson's disease in humans or non-human mammals; 4) Application in the preparation of products for screening or assisting screening of heart failure caused by Parkinson's disease in humans or non-human mammals.

2. Use of a product for detecting NTN1 protein, RELN protein and / or SLIT1 protein or mRNA or mRNA fragments of their transcription products in any of the following: 1) Application in the preparation of products for diagnosing or assisting in the diagnosis of Parkinson's disease in humans or non-human mammals; 2) Application in the preparation of products for screening or assisting screening of Parkinson's disease in humans or non-human mammals; 3) Application in the preparation of products for diagnosing or assisting in the diagnosis of heart failure caused by Parkinson's disease in humans or non-human mammals; 4) Application in the preparation of products for screening or assisting screening of heart failure caused by Parkinson's disease in humans or non-human mammals.

3. The use according to claim 2, characterized in that: The product for detecting NTN1 protein, RELN protein and / or SLIT1 protein or their transcription product mRNA or mRNA fragments includes a product for detecting the expression level of NTN1 protein, RELN protein and / or SLIT1 protein or their transcription product mRNA or mRNA fragments in a sample to be tested; And / or, the product for detecting NTN1 protein, RELN protein and / or SLIT1 protein or their transcription product mRNA or mRNA fragments includes a kit, a reagent, a test strip, a primer, a probe or a chip; And / or, the sample to be tested is cardiac tissue or blood of a human or non-human mammal; optionally, selected from blood.

4. The use according to claim 3, characterized in that The reagents include: antibodies, proteins and / or polypeptides that specifically bind to NTN1 protein, RELN protein and / or SLIT1 protein; or reagents that detect mRNA or mRNA fragments of NTN1 protein, RELN protein and / or SLIT1 protein transcription products.

5. The use according to claim 4, characterized in that The antibody, protein and / or polypeptide is modified with an isotope, horseradish peroxidase, colloidal gold or a fluorescent probe, or is unmodified; And / or, the reagent for detecting the mRNA or mRNA fragment of the transcription product of NTN1 protein, RELN protein and / or SLIT1 protein includes a PCR reaction buffer or a nucleic acid molecule hybridization buffer; optionally, it includes a specific sequence fragment of the gene fragment of NTN1 protein, RELN protein and / or SLIT1 protein or its mRNA, or a molecular probe containing these specific sequence fragments.

6. A product for diagnosing, assisting in the diagnosis, screening or assisting in the screening of Parkinson's disease or Parkinson's disease-induced heart failure in humans or non-human mammals, characterized in that: It includes products for detecting the expression level of NTN1 protein, RELN protein and / or SLIT1 protein or their transcription product mRNA or mRNA fragment in the sample to be tested.

7. The product for diagnosis, auxiliary diagnosis, screening or auxiliary screening of Parkinson's disease or Parkinson's disease-induced heart failure in humans or non-human mammals according to claim 6, characterized in that: The products for detecting NTN1 protein, RELN protein and / or SLIT1 protein or their transcription products mRNA or mRNA fragments include kits, reagents, test strips, primers, probes or chips; And / or, the sample to be tested is cardiac tissue or blood of a human or non-human mammal; optionally, selected from blood.

8. A device for diagnosing, assisting in diagnosis, screening or assisting in screening Parkinson's disease or Parkinson's disease-induced heart failure in humans or non-human mammals, characterized in that: include: A detection unit, used to obtain the expression level of NTN1 protein, RELN protein and / or SLIT1 protein or their transcription product mRNA or mRNA fragment in the sample to be tested; The judgment unit is used to diagnose or assist in diagnosis, screening or assist in screening whether a human or non-human mammal suffers from Parkinson's disease or Parkinson's-induced heart failure based on the expression level of NTN1 protein, RELN protein and / or SLIT1 protein or their transcription product mRNA or mRNA fragment in the test sample.

9. The device for diagnosing, assisting in diagnosis, screening or assisting in screening Parkinson's disease or Parkinson's disease-induced heart failure in humans or non-human mammals according to claim 8, characterized in that: Compared with the expression amount of NTN1 protein, RELN protein and / or SLIT1 protein or their transcription product mRNA or mRNA fragments in the sample of a normal human or normal non-human mammal, the expression amount of NTN1 protein, RELN protein and / or SLIT1 protein or their transcription product mRNA or mRNA fragments in the sample of the human or non-human mammal to be tested is reduced, and the human or non-human mammal to be tested is judged to suffer from Parkinson's disease or Parkinson's disease-induced heart failure.

10. A method for screening drugs for treating Parkinson's disease or Parkinson's disease-induced heart failure, characterized in that: include: The candidate drugs are used in the non-human mammal model of Parkinson's disease or Parkinson's disease induced heart failure; the drugs that up-regulate the expression of NTN1 protein, RELN protein and / or SLIT1 protein or their transcription product mRNA or mRNA fragments or enhance their activity are screened, or the candidate drugs that alleviate Parkinson's disease or Parkinson's disease induced heart failure are screened as the drugs for treating Parkinson's disease or Parkinson's disease induced heart failure.

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