CN117805249A - Biomarker for diagnosis of depression and application thereof - Google Patents
Biomarker for diagnosis of depression and application thereof Download PDFInfo
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
- G01N30/62—Detectors specially adapted therefor
- G01N30/72—Mass spectrometers
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Abstract
The invention discloses an application of a compound 4-TMAP as a biomarker for depression diagnosis; the invention discovers that the difference of the 4-TMAP in the serum or the feces of the patients with the depression and the healthy people has statistical significance, and can conveniently prompt whether the patients with the depression have the depression or prompt the potential depression risk by analyzing the relative content of the 4-TMAP in the serum or the feces of the patients to be detected. The detection method is simple and convenient, is convenient for physical examination application and large-scale screening, is favorable for early detection and early treatment, and has wide market prospect.
Description
Technical Field
The invention relates to the field of disease diagnosis, in particular to a biomarker related to pathogenesis of depression and application thereof.
Background
Depression (MDD) is now the most common psychological disorder, with continuous and long-term mood depression as the major clinical feature, the most important type of modern human psychological disorder. Clinically, the mood is low and the reality is too careless, the mood is low and subsides for a long time, from the beginning of smoldering to the last of sadness, spewing, pain, pessimistic and aversion, the feeling of living is hopefully afflicting itself every day, negatively, evading, and finally, the feeling of suicidal tendency and behavior is even more. The patient suffers from somatic symptoms. Chest oppression and shortness of breath.
Depression (MDD) is a common disabling disease that is highly developed worldwide and has a significant impact on the physical and mental health of patients. Depression can lead to impaired working and learning abilities of the patient, severely affecting their quality of life, and possibly causing their suicide. A meta analysis of global disease burden study in 2017 indicated that depression has affected about 3 million people and is one of the leading causes of disability and disease burden worldwide.
Diagnosis of depression should be based mainly on medical history, clinical symptoms, course of disease, physical examination and laboratory examination, and diagnosis of typical cases is generally not difficult. The internationally common diagnostic criteria are typically ICD-10 and DSM-IV. ICD-10 is mainly adopted in China, and refers to first-onset depression and recurrent depression, and bipolar depression is not included. Patients often have typical symptoms such as low mood, loss of interest and pleasure, poor vigor or tiredness. Other common symptoms are (1) reduced ability to focus on and pay attention; (2) a decrease in self-assessment; (3) Self-criminal ideas and worthlessness (even in mild attacks); (4) consider the prospect to be dull and pessimistic; (5) ideas or behaviors of self-injury or suicide; (6) sleep disorders; (7) appetite decrease.
Proper diagnosis, especially early diagnosis of depression, is critical to effective treatment; however, the diagnosis of depression is mainly based on clinical symptomatology standards, and the clinical symptoms of depression are various and lack of specificity, so that the misdiagnosis rate is high, and effective treatment is hindered. Unfortunately, while researchers have been working on this for decades, many biomarkers associated with the pathogenesis of depression have also been discovered, unfortunately, there is still a lack of clinically effective diagnostic markers.
Disclosure of Invention
The invention mainly aims to provide a biomarker related to depression, which can effectively solve the problems in the background technology.
In order to achieve the above purpose, the technical scheme adopted by the invention is as follows:
there is provided the use of a compound 4-TMAP of formula (I) as or for the preparation of a biomarker for the diagnosis of depression, the compound 4-TMAP having the structure
Further, the invention provides a detection kit, which comprises the compound 4-TMAP and necessary diluent; preferably, the 4-TMAP and diluent may be provided in a concentration of (3-10) ng/ml, preferably 4-8ng/ml, more preferably 5ng/ml; preferably the diluent comprises water.
Furthermore, the invention provides a depression early warning system, which comprises the detection kit and an analysis tool, wherein the analysis tool preferably comprises High Performance Liquid Chromatography (HPLC) or high performance liquid chromatography-mass spectrometry (HPLC-MS).
Further, the present invention provides a method for diagnosing depression, said method comprising the step of determining the level of 4-TMAP in serum or stool of a human or animal subject; preferably, the prompting or early warning is carried out when the content of the 4-TMAP is more than or equal to 5 ng/ml.
Compared with the prior art, the invention has the following beneficial effects:
1. the invention discovers the statistical difference of the relative content of 4-TMAP in the feces and serum of healthy people and patients with depression for the first time, and determines the application of the 4-TMAP as a biomarker for diagnosis of depression.
2. The method can conveniently and quickly screen depression patients or potential depression patients in early stage, discover depression risks, facilitate early discovery and early treatment, and prevent the deterioration of the conditions of the potential depression patients.
Drawings
Fig. 1: the relative content of 4-TMAP in serum samples from depressed patients (MDD) and healthy persons (HC);
fig. 2: the relative content of 4-TMAP in faecal samples of depressed patients (MDD) and healthy persons (HC);
fig. 3: the relative amounts of 4-TMAP in serum samples of normal food group and amino acid food group in AFD-induced mouse depression model;
fig. 4: the relative amounts of 4-TMAP in the fecal samples of the normal food group and the amino acid food group in the AFD-induced depression model of mice;
fig. 5: relative content of 4-TMAP in serum samples of normal mice and CRS depressed mice in a CRS-induced mouse depression model;
fig. 6: relative content of 4-TMAP in fecal samples of normal mice and CRS-depressed mice in CRS-induced mouse depression model;
fig. 7: the relative content of 4-TMAP in serum samples of normal mice and Cort depressed mice in Cort induced mouse depression model;
fig. 8: the relative content of 4-TMAP in fecal samples of normal mice and Cort depressed mice in Cort induced mouse depression model;
fig. 9: distribution of 4-TMAP in intestinal and whole brain samples of normal mice, AFD, CRS, cort-induced mouse depression model.
Detailed Description
The invention is further described in connection with the following detailed description, in order to make the technical means, the creation characteristics, the achievement of the purpose and the effect of the invention easy to understand.
Example 1:4-TMAP (4-TMAP) can be used as clinical diagnosis marker of depression
Faecal and serum samples from depressed patients (MDD) and healthy persons (HC) were collected and the level of 4-TMAP was detected by HPLC-MS mass spectrometry as follows:
(1) Sample treatment and extraction:
taking 100 mu L of serum sample, adding acetonitrile to fix volume to 1mL, swirling for 1min, mixing uniformly, and centrifuging for 10min at 12000 r/min. The supernatant solution was filtered through a 0.22 μm filter and injected by HPLC-MS.
Taking 100mg of fecal sample, adding acetonitrile to a volume of 1mL, swirling for 1min, uniformly mixing, and centrifuging for 10min at 12000 r/min. The supernatant solution was filtered through a 0.22 μm filter and injected by HPLC-MS.
Preparing a standard substance solution: taking standard substance mother solution, adding water to prepare standard substance solutions with the concentration of 1ng/mL, 2ng/mL, 5ng/mL, 10ng/mL, 20ng/mL, 50ng/mL and 100ng/mL respectively, and filtering with a 0.22 mu m filter membrane, and carrying out HPLC-MS/MS analysis.
(2) The detection parameters are as follows:
HPLC parameters:
liquid chromatography model: LC30
Chromatographic column: waters ACQUITY UPLC HILLC (100 mm. Times.2.1 mm,1.7 μm)
Mobile phase: column temperature of 0.1% formic acid aqueous solution: 35 DEG C
And B, methanol flow rate: 0.30mL/min
Sample injection volume: 1 mu L
Gradient elution procedure:
mass spectrometry conditions:
mode: ESI, positive ion mode analysis, scan mode: multiple Reaction Monitoring (MRM), dry gas: nitrogen 10.0L/min, heating gas: air 10.0L/min, collision gas: argon 270kPa, atomizing gas: 3.0L/min, interface temperature: 200 ℃, DL tube temperature: 280 ℃, residence time: 150ms, heating module temperature: 300 ℃, delay time: 3ms, interface voltage: 1.0KV, and both Q1 and Q3 are unit resolution liquid instrument models: shimadzu LC-MS-MS-8050
TABLE 2 multiple reaction monitoring mode (MRM) parameters
Tab.2 Multiple reaction monitoring(MRM)conditions
Detection conclusion:
as shown in fig. 1 and 2, the level of 4-TMAP in the MDD (n=142) serum samples was significantly lower than HC (n=49), which is statistically significant; the level of 4-TMAP in the MDD (n=114) stool samples was significantly lower than HC (n=47), suggesting that 4-TMAP has potential as a clinical diagnostic marker for depression.
When a fecal or serum sample from a patient to be tested is treated and tested by the above method, wherein the 4-TMAP content is below 5ng/mg or 5ng/ml, a higher risk of depression is indicated, indicating that further retesting according to other means is required to determine whether depression is present.
Example 2:4-TMAP is significantly reduced in AFD-induced mouse depression model
The 6 SPF grade C57/B6j male mice (purchased from Jieqiao medicine) which are born for 2 days are randomly divided into 2 groups, one group is a normal food group (NCD, 3) and one group is an amino acid food group (AFD, 3), and each group is matched with 1 lactating female mouse, and the female mice respectively eat corresponding foods and then lactating mice. After 4 weeks of continuous feeding, female mice were isolated, the amino acid diet group was changed to normal diet, and their depressed status was monitored by forced swimming, tail suspension, sugar water preference experiments.
Serum, stool, whole brain and intestinal tissues of NCD and AFD mice, respectively, were taken and assayed for 4TMAP levels by HPLC-MS.
Serum and stool treatment and detection methods are the same as those of patient samples. The results of the assay are shown in FIG. 3 and FIG. 4, where 4-TMAP was significantly reduced in serum and fecal samples from AFD-induced mouse models of depression.
The detection method of the whole brain and intestinal tract comprises the following steps: the fresh whole brain and intestinal tract of the mice were removed, snap frozen in liquid nitrogen, frozen sections, 10um, and the distribution of 4-TMAP was detected by imaging mass spectrometry.
Example 3:4-TMAP is significantly reduced in CRS-induced mouse depression model
6 SPF class C57/B6j male mice (purchased from JieXtensible medicine) 6-8 weeks old were randomly divided into 2 groups, one group being normal mice (3) and one group being CRS mice (3). After acclimation, from day 0, the CRS mice group received chronic, unpredictable mild stimulation over a period of 21d, mimicking chronic low-intensity stress received in human daily life. One stimulus was selected from each of the daytime and nighttime daily and mice were treated in a random, continuous, non-repetitive sequence, so that the mice could not predict the appearance of the stimulus. After continuous modeling 21d, the success of depression modeling (significant differences from normal mice) was confirmed by forced swimming, tail suspension, and sugar water preference experiments.
Serum, faeces, whole brain and intestinal tissues of NCD and CRS mice, respectively, were taken and the level of 4TMAP was measured by mass HPLC-MS.
Serum and stool treatment and detection methods are the same as those of patient samples. The test results are shown in fig. 5 and 6, and 4-TMAP was significantly reduced in serum and fecal samples of CRS-induced mouse depression model.
The detection method of the whole brain and intestinal tract comprises the following steps: the fresh whole brain and intestinal tract of the mice were removed, snap frozen in liquid nitrogen, frozen sections, 10um, and the distribution of 4-TMAP was detected by imaging mass spectrometry.
Example 4:4-TMAP is significantly reduced in Cort-induced mouse models of depression
6 SPF grade C57/B6j male mice (purchased from JieXtensible medicine) 6-8 weeks old were randomly divided into 2 groups, one group being normal mice (3) and one group being Cort (3). After acclimatization, from day 0, cort mice group (MCE, HY-B1618) received drinking water as a corticosterone solution at a final concentration of 25 μg/mL (pH 7.0-7.4), normal mice group received plain drinking water, changed every 2d, and molded 21d continuously. After the modeling is finished, modeling success (obvious difference from normal mice) is confirmed through forced swimming, tail suspension and syrup preference experiments
Serum, faeces, whole brain and intestinal tissues of NCD and Cort mice, respectively, were taken and assayed for 4TMAP levels by HPLC-MS.
Serum and stool treatment and detection methods are the same as those of patient samples. The results of the assay are shown in FIG. 7 and FIG. 8, where 4-TMAP was significantly reduced in serum and fecal samples from Cort-induced mouse models of depression.
The detection method of the whole brain and intestinal tract comprises the following steps: the fresh whole brain and intestinal tract of the mice were removed, snap frozen in liquid nitrogen, frozen sections, 10um, and the distribution of 4-TMAP was detected by imaging mass spectrometry.
The results of the imaging mass spectra are shown in figure 9,4-TMAP was significantly reduced in both intestinal and whole brain samples of AFD, CRS, cort induced mouse depression models.
The foregoing has shown and described the basic principles and main features of the present invention and the advantages of the present invention. It will be understood by those skilled in the art that the present invention is not limited to the embodiments described above, and that the above embodiments and descriptions are merely illustrative of the principles of the present invention, and various changes and modifications may be made without departing from the spirit and scope of the invention, which is defined in the appended claims. The scope of the invention is defined by the appended claims and equivalents thereof.
Claims (10)
1. Use of a compound 4-TMAP shown in formula (I) for preparing a biomarker for diagnosing depression, wherein the structure of the compound 4-TMAP is as follows
2. A test kit, characterized in that said kit comprises the biomarker of claim 1.
3. The test kit according to claim 2, wherein the test kit further comprises a necessary diluent.
4. The kit of claim 3, wherein the biomarker and diluent are configured at a concentration of 3-10 ng/mL.
5. The kit of claim 3, wherein the biomarker and diluent are configured to a concentration of 4-8 ng/mL.
6. The test kit of claim 3, wherein the biomarker and diluent are configured at a concentration of 5 ng/mL.
7. The test kit according to any one of claims 3 to 6, wherein the diluent is selected from the group consisting of water.
8. A depression pre-warning system, characterized in that it comprises a detection kit and an analysis tool according to any one of claims 2-7.
9. The depression pre-warning system of claim 8, wherein the analytical tool is selected from the group consisting of High Performance Liquid Chromatography (HPLC).
10. The depression pre-warning system of claim 8, wherein the analytical tool is selected from the group consisting of high performance liquid chromatography-mass spectrometry (HPLC-MS).
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202211165755.8A CN117805249A (en) | 2022-09-23 | 2022-09-23 | Biomarker for diagnosis of depression and application thereof |
| PCT/CN2023/119893 WO2024061250A1 (en) | 2022-09-23 | 2023-09-20 | Diagnostic biomarker for depression and use thereof |
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| CN202211165755.8A CN117805249A (en) | 2022-09-23 | 2022-09-23 | Biomarker for diagnosis of depression and application thereof |
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| MX2009006007A (en) * | 2006-12-05 | 2009-07-17 | Neurogesx Inc | Prodrugs and methods of making and using the same. |
| EP2513653A1 (en) * | 2009-10-09 | 2012-10-24 | Carolyn Slupsky | Methods for diagnosis, treatment and monitoring of patient health using metabolomics |
| WO2018150077A1 (en) * | 2017-02-16 | 2018-08-23 | Afekta Technologies Oy | Betaine biomarkers for nutritional status, nutritional regimen, endogenous metabolism, or risk of cardiovascular or metabolic diseases |
| US20210353611A1 (en) * | 2018-09-20 | 2021-11-18 | Yeda Research And Development Co. Ltd. | Methods of treating amyotrophic lateral sclerosis |
| WO2020226683A1 (en) * | 2019-05-07 | 2020-11-12 | The Regents Of The University Of California | Compositions and methods for promoting healthy neural development in an unborn baby |
| EP3879272A1 (en) * | 2020-03-09 | 2021-09-15 | Albert-Ludwigs-Universität Freiburg | Gut microbiota-related methods for treating dementia and age-dependent cognitive decline |
| JP2023508968A (en) * | 2019-12-23 | 2023-03-06 | アルベルト・ルートヴィヒ大学フライブルク | Gut Microbiota-Related Methods for Treating Dementia and Age-Dependent Cognitive Decline |
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