CN115561463A - Serum VEGFB level as mood disorder-related disease marker and application thereof - Google Patents

Abstract

The invention provides a novel marker for symptoms and diseases related to mood disorder, in particular depression and schizophrenia. The VEGFB level as a marker can be used for preparing products for diagnosing diseases related to the emotional disorders and/or evaluating risks of the diseases related to the emotional disorders and related medicaments.

Description

Serum VEGFB level as mood disorder-related disease marker and application thereof

Technical Field

The invention belongs to the technical field of biological medicines, and particularly relates to a potential marker for detecting the level of VEGFB in serum as anxiety and depression and application thereof.

Background

Depression is a common mental disorder, characterized clinically by marked and persistent mood depression and diminished interest. Depression is a psychiatric disease with a high incidence in china and even the world, and it is a major factor that contributes to the global burden of disease. Patients with Major Depressive Disorder (MDD) have a significantly increased risk of suicide compared to the general population. Like other psychiatric disorders, the etiology of depression is complex and there is a lack of specific diagnostic tools for the disease. The clinical diagnosis using the mental analysis scale is affected by the subjective consciousness of the patient and depends on the judgment of the doctor, so that the diagnosis is prone to be deviated.

In addition, due to the specificity of psychiatric disorders, pathological diagnosis by brain biopsy is not possible. Therefore, there is a clinical need for an objective and feasible diagnostic method to enhance the prevention and treatment of depression and improve the quality of life of patients. Peripheral blood sampling is noninvasive, safe and simple, and is expected to be used as a substitute for brain tissue biopsy for diagnosis of central nervous system diseases such as depression. Peripheral blood gene markers are widely researched in the field of depression, but the use of the peripheral blood gene markers as a diagnosis of depression also faces problems that the markers obtained by different researches are different, a diagnosis model does not have universality, and clinical diagnosis is different.

Disclosure of Invention

VEGF-B is one of the vascular endothelial growth factors and has been found to have therapeutic effects in oxidative stress related diseases. Nevertheless, the knowledge of the antioxidant VEGF-B is still limited.

In the present study, the inventors screened serum of young men performing a task under long-term stress for potential markers of depression by protein chips and found that serum levels of VEGFB were reduced after performing the task compared to before performing the task. To further prove the relation between VEGFB and depression, the inventor establishes a classical mouse anxiety and depression induction model, analyzes the change of serum VEGFB level in mice in the model through ELISA detection, and finds that the VEGFB level does not change obviously in a control mouse but drops obviously in the anxiety and depression mouse, and prompts that the serum VEGFB level can be a peripheral marker of depression. Furthermore, the inventor discovers that the VEGFP level in the serum of patients with anxiety depression syndrome is remarkably reduced compared with healthy controls through clinical diagnosis results of patients with depression in different regions and patients with anxiety depression syndrome, schizophrenia, alzheimer disease and vascular dementia, and further proves that VEGFB can be used as a marker for diagnosing depression and/or evaluating the risk of depression, thereby completing the invention.

In one aspect, the invention provides the use of VEGFB as a marker for the preparation of a product or medicament for diagnosing a symptom disease associated with a mood disorder and/or assessing the risk of developing a symptom disease associated with a mood disorder.

VEGFB can be used as a detection object, preferably serum VEGFB, and the expression in a blood sample or a serum sample is reduced or the level of the VEGFB is reduced, and products comprise a chip or a kit.

The symptom disease associated with mood disorders in the present invention is preferably depression or schizophrenia.

The application of VEGFB in screening drugs for treating diseases and/or relieving symptoms related to the emotional disorders is disclosed.

VEGFB can be used as an object of detection, and serum VEGFB is preferable. Down-regulated expression or reduced levels in a blood sample or serum sample of an individual suffering from or at high risk for developing a symptomatic disease associated with a mood disorder.

According to the present invention, a downregulation or a reduction in the level of VEGFB expression in a test subject by at least 30% or more, such as at least 40%, at least 50% or at least 60%, compared to the level of VEGFB in the same subject not diagnosed with depression or the average VEGFB level in a group of healthy subjects may be predictive for a test subject suffering from depression or having a high risk of suffering from depression.

According to the invention, the product comprises a chip or a kit. The kit comprises, but is not limited to, an ELISA detection kit, a colloidal gold detection kit, a co-immunoprecipitation kit, a chemiluminescence kit, an immunofluorescence kit and the like. These kits may be prepared according to conventional methods in the art. For example, an ELISA detection kit comprises: solid phase carrier for coating antibody specifically combined with serum VEGFB, enzyme labeled antibody, enzyme substrate, VEGFB standard substance, negative reference substance, diluent, washing liquid, enzyme reaction stopping liquid and the like. Wherein, the antibody specifically binding with serum VEGFB can be VEGFB monoclonal antibody.

In another aspect, the invention provides the use of VEGFB for screening a medicament for the treatment of depression and/or alleviation of symptoms of depression.

According to the present invention, the depression includes mild, moderate and severe depression, postpartum depression, adolescent depression, adult depression, senile depression, and the like. The depression symptoms include symptoms of mood depression, decreased interest, disuse, autonomy, thought retardation, slow behavior, cognitive function impairment, hypomnesis, attention disorder, sleep disorder, anorexia, hyposexuality, etc. of a patient diagnosed with depression.

According to the invention, the medicament can be any clinically suitable dosage form, including but not limited to tablets, capsules, granules, dripping pills, injections, oral liquid and the like.

Advantageous effects

The inventors have discovered a new potential marker for depression, namely serum VEGFB; serum VEGFB levels were significantly reduced in a classical depressed mouse model. Aiming at the potential marker, the inventor verifies through clinical experiments of patients with different anxiety and depression syndromes in different regions, thereby not only verifying that VEGFB can be used as a marker of depression, but also verifying that VEGFB is mainly related to diseases related to mood disorders, and having clinical identification and diagnosis significance for schizophrenia.

Drawings

FIG. 1 shows the protein chip results.

Fig. 2 shows the ELISA results of model mouse serum VEGFB levels.

Fig. 3 shows the results of ELISA of serum VEGFB levels from clinical tests of patients with beijing and sichuan depression.

Figure 4 shows the clinical diagnosis of anxio-depressive syndrome with patients with schizophrenia, alzheimer's disease and vascular dementia.

Detailed Description

The present invention will be described in further detail with reference to specific examples. It is to be understood that the following examples are only illustrative and explanatory of the present invention and should not be construed as limiting the scope of the present invention. All the technologies realized based on the above-mentioned contents of the present invention are covered in the protection scope of the present invention.

Unless otherwise indicated, the raw materials and reagents used in the following examples are all commercially available products or can be prepared by known methods.

Example 1

1. Materials and methods (I) high throughput cytokine chip screening

For young men facing higher mental stress in a long-term task, peripheral venous blood was collected before and after the task, wherein 8 samples before the task and 12 samples after the task were collected. The AAH-BLG-1 chip of Raybiotech company is used for screening target proteins from 507 factors. The readout is the fluorescence intensity, and after normalization, higher values indicate higher protein concentrations.

(II) statistical analysis

Statistical analysis of all data was performed using GraphPad Prism 6.01 and SPSS19.0 software. Data results are expressed as Mean ± standard deviation (Mean ± SD), mean between groups is statistically analyzed by t-test, P <0.05 is considered statistically significant

2. Results

1. The protein chip screening results (fig. 1) demonstrate that VEGFB levels are reduced in sera of young men performing tasks under long-term stress. p =0.027342, statistically significant.

Example 2

1. Materials and methods

A binding device is made by chiseling small ventilation holes on the tube wall and the tube cover of a 50ml pointed-bottom centrifuge tube, the hole wall is smooth as much as possible, and unnecessary damage to mice is avoided. The mouse in the bound group is placed into a centrifuge tube, the tail penetrates out of a central vent hole of a tube cover, the tube cover is screwed to limit the movement of the mouse, the centrifuge tube is inclined at 45 degrees, and the head of the mouse is enabled to face upwards. Restraint stress at a fixed time period of each day 8:00 to 14:00, washing the centrifugal tube after finishing the operation, ensuring that the centrifugal tube is clean and has no excrement and odor residue, and naturally drying the centrifugal tube and then recycling the centrifugal tube for later use. Restraint stress lasted for a total of 8 days. Then blood is collected from the orbital venous plexus.

Detection was carried out using mouse VEGFB ELISA kit (cat # EM 0216) of Wuhan Feien, inc. according to the instructions.

Statistical analysis of all data was performed using GraphPad Prism 6.01 and SPSS19.0 software. Data results are expressed as means ± standard deviation (Mean ± SD), mean values between groups are statistically analyzed by t-test, and differences are considered statistically significant when P < 0.05.

2. As a result, the

Binding of mice for 6 hours a day for 8 days was a classical model of inducing anxiety in mice, and serum VEGFB levels in mice after binding were found to be significantly reduced by ELISA assay (p = 0.000104) (fig. 2), suggesting that serum VEGFB levels may be a peripheral marker of depression.

Example 3

1. Materials and methods

23 depression anxiety syndrome patients who visit the first medical center of the general hospital of the liberation force from 2019 to 2019 and 8 are collected, and 20 patients are compared with healthy volunteers which are from the first medical center of the general hospital of the same liberation force and have normal indexes. 19 patients with depression anxiety syndrome who were admitted to hospital in the fourth people's hospital in Sichuan province city, 10/23/2020 were selected. Collecting 5ml of elbow median venous blood of all the subjects, standing at room temperature for 30min, centrifuging at 1000 Xg for 15min, subpackaging and freezing in a refrigerator of-70 deg.C.

Human serum VEGFB level detection, because of lack of sensitive ELISA kit, human serum verification utilizes a customized AAH-BLG-1 chip of Raybiotech company, and the level of VEGFB in human serum is specially analyzed.

Statistical analysis of all data was performed using GraphPad Prism 6.01 and SPSS19.0 software. Data results are expressed as Mean ± standard deviation (Mean ± SD), mean between groups is statistically analyzed by t-test, and differences are considered statistically significant with P < 0.05.

2. Results

It was determined that patients with either the anxiety-depressive syndrome in Beijing or Sichuan presented decreased levels of serum VEGFB, with significant differences between Beijing patients and healthy controls (p = 0.001019) and Sichuan patients and healthy controls (p = 0.012789). There was no statistical difference between beijing and sichuan patients (p = 0.997859). Specific results are shown in FIG. 3.

Example 4

1. Materials and methods

23 depression anxiety syndrome patients who visit the first medical center of the general hospital of the liberation force from 2019 to 2019 and 8 are collected, and 20 patients are compared with healthy volunteers which are from the first medical center of the general hospital of the same liberation force and have normal indexes. 19 patients with depression anxiety syndrome, 20 patients with schizophrenia, 13 patients with Alzheimer disease and 6 patients with vascular dementia who are hospitalized in the fourth people hospital of Sichuan province city, sichuan province, 10 months and 23 days in 2020 were selected. Collecting 5ml of elbow median venous blood of all the subjects, standing at room temperature for 30min, centrifuging at 1000 Xg for 15min, subpackaging and freezing in a refrigerator of-70 deg.C.

Human serum VEGFB level detection, because of lack of sensitive ELISA kit, human serum verification utilizes a customized AAH-BLG-1 chip of Raybiotech company, and the level of VEGFB in human serum is specially analyzed.

Statistical analysis of all data was performed using GraphPad Prism 6.01 and SPSS19.0 software. Data results are expressed as means ± standard deviation (Mean ± SD), mean values between groups are statistically analyzed by t-test, and differences are considered statistically significant when P < 0.05.

2. Results

When Beijing and Sichuan patients were combined into one group, the VEGFP levels in serum of patients with anxiety-depression syndrome were significantly reduced compared to healthy controls (p = 000356). Serum VEGFB levels in schizophrenic patients were also significantly reduced compared to healthy controls (p = 000338), with no significant difference from patients with anxiety-depression syndrome (p = 482373). Serum VEGFB levels in patients with alzheimer's disease and vascular dementia were close (p = 0.933026) because both groups were less numerous, and combining both groups into a dementia group, the dementia patients were statistically not different from healthy controls (p = 0.209745), significantly different from patients with anxiety depression syndrome (p = 0.012426). Therefore, VEGFB has certain differential diagnosis significance, and is suggested to be mainly related to emotional disorder. Specific results are shown in FIG. 4.

The embodiments of the present invention have been described above. However, the present invention is not limited to the above embodiment. Any modification, equivalent replacement, or improvement made within the spirit and principle of the present invention should be included in the protection scope of the present invention.

Claims (10)

1. Use of vegfb as a marker for the preparation of a product or medicament for diagnosing a symptom disease associated with a mood disorder and/or assessing the risk of developing a symptom disease associated with a mood disorder.
2. 2. Use according to claim 1, wherein the VEGFB is the subject of detection, preferably serum VEGFB.
3. 3. Use according to any one of claims 1 or 2, wherein the VEGFB is down-regulated in expression or reduced in level in a blood sample or serum sample of a mood disorder-related symptom disorder or of an individual at high risk of developing a mood disorder-related symptom disorder.
4. 4. The use of any one of claims 1-3, wherein the product comprises a chip or a kit.
5. 5. Use according to claims 1-4, wherein the symptomatic disease associated with mood disorders is depression or schizophrenia.
6. Use of vegfb in the screening of a medicament for treating a disease associated with a symptom of a mood disorder and/or alleviating a symptom associated with a mood disorder.
7. 7. Use according to claim 6, wherein the VEGFB is to be tested, preferably serum VEGFB.
8. 8. Use according to any one of claims 6 or 7, wherein the VEGFB is down-regulated in expression or reduced in level in a blood sample or serum sample of a mood disorder-related symptom disorder or of an individual at high risk of developing a mood disorder-related symptom disorder.
9. 9. The use of any one of claims 6-8, wherein the medicament can be in any clinically suitable dosage form, including but not limited to, tablets, capsules, granules, drop pills, injections, oral liquids, and the like.
10. 10. Use according to claims 6-9, wherein the symptomatic disease associated with mood disorders is depression or schizophrenia.

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