CN112649608A - Application of MMP19 in serum in anxiety and depression - Google Patents
Application of MMP19 in serum in anxiety and depression Download PDFInfo
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Abstract
The invention relates to the field of biological detection, in particular to application of MMP19 in serum in anxiety and depression. The invention provides an application of MMP19 protein or a specific antibody thereof in preparation of products for detecting or diagnosing anxiety and depression. The invention uses the serum MMP19 protein as a new marker for screening drugs and diagnosing anxiety and depression. The invention provides a kit for preparing and diagnosing anxiety and depression aiming at MMP19 for the first time, and the anxiety and depression can be screened and diagnosed only by detecting the concentration of MMP-19 in sample serum, so that the diagnosis efficiency and accuracy are improved, and an important value is provided for early diagnosis of anxiety and depression. The composition is of great help to the prevention and treatment of anxiety and depression in China.
Description
Technical Field
The invention relates to the field of biological detection, in particular to application of MMP19 in serum in anxiety and depression.
Background
Depression belongs to the group of affective disorders, and is mood disorder or affective disorder caused by various factors. The main manifestations are depressed mood, thought retardation, hypovolemia, cognitive impairment, loss of interest and ability in activity, loss of self-esteem, inappropriate feelings of guilt, death and suicidal thoughts, reduced concentration, sleep disturbance and anorexia, hyposexuality, and may be accompanied by various somatic symptoms.
China is in the transformation period of rapid development, the living and working pressure of urban and rural residents is continuously increased, and the social and psychological problems are increasingly prominent. The survey report of health organization shows that about 17.5 percent of adults over 18 years old in China suffer from various mental and psychological disorders, and the cardiovascular diseases and tumors are the main causes of the burden of Chinese diseases. Wherein, the incidence rate of the depression is obviously increased, and the national survey in 1982 shows that the prevalence rate of the depression is only 0.76%; according to the global depression epidemiology reported by Nature in 2014, 3.5 hundred million people suffer from depression in the global range, and the prevalence rate of depression in China is 3.02%; increased by about 40 times in thirty years. The symptoms of depression are very heterogeneous, and specific and sensitive biomarkers are needed for differential diagnosis and treatment response judgment, but related markers are relatively lacking.
Matrix Metalloproteinases (MMPs) are a family of proteolytic enzymes that degrade almost all components of the extracellular matrix. MMP19(matrix metalloproteinase 19) is one of the MMPs family members, and is first isolated from autoimmune rheumatoid arthritis and then sequentially found in various tissues of the body, and the expression degree thereof varies. At present, no association of MMP-19 with depression has been reported.
Disclosure of Invention
In order to solve the problems, the invention aims to provide a diagnostic serum marker MMP19 for anxiety depression and a product application thereof.
In order to achieve the purpose, the specific technical scheme of the invention is as follows:
the invention firstly provides the application of MMP19 protein or a specific antibody thereof in the preparation of products for detecting or diagnosing anxiety and depression.
In some embodiments, the sample for detecting anxious depression is serum.
Relative expression levels of MMP19 protein were up-regulated in serum of patients with anxiety depression relative to healthy humans.
In some embodiments, the product comprises an early diagnostic reagent or kit for detecting anxio-depressive disorder.
Further, the invention provides a diagnostic kit for detecting anxiety and depression, which comprises a reagent for detecting the content of MMP19 protein in serum.
In some embodiments, the reagent for detecting the content of the MMP19 protein in the serum is an antibody specific to MMP19 protein.
In some embodiments, the kit further comprises an anti-human MMP19 protein antibody coated enzyme-labeled reaction plate, a specimen diluent, a washing solution, an enzyme-labeled antigen, a developing solution, a stop solution, a positive control and a negative control.
Furthermore, the invention provides an application of the MMP19 protein in screening of candidate drugs for preventing or treating anxiety and depression.
In some embodiments, the candidate drug downregulates the expression level of MMP19 protein in the blood.
In some embodiments, the drug candidate comprises an antibody to the MMP19 protein and/or an inhibitor of the MMP19 protein.
Based on the technical scheme, the invention has the following beneficial effects:
experiments prove that compared with normal human serum, MMP19 protein is up-regulated in serum of patients with depression and anxiety; while diabetic peripheral neuropathy and cerebral infarction were indistinguishable from controls, demonstrating that serum MMP-19 levels are not elevated in all neuropathies, specific for the diagnosis of anxiety depression.
The invention takes the serum MMP19 as a new marker for screening drugs and diagnosing anxiety and depression. The invention provides a kit for preparing and diagnosing anxiety and depression aiming at MMP19 for the first time, and the anxiety and depression can be screened and diagnosed only by detecting the concentration of MMP-19 in sample serum, so that the diagnosis efficiency and accuracy are improved, and an important value is provided for early diagnosis of anxiety and depression. The composition is of great help to the prevention and treatment of anxiety and depression in China.
Drawings
FIG. 1 the chip detects MMP-19 changes in serum before and after the performance of a high pressure long-term task;
FIG. 2ELISA detection of MMP-19 levels in serum of anxiety-depressed patients and healthy volunteers;
FIG. 3ELISA detection of MMP-19 levels in serum of diabetic peripheral neuropathy, cerebral infarction patients and healthy volunteers;
FIG. 4 calculated ROC curves using ELISA results, where Depression group area0,867, Std.0.05754, 95% 0.7543to 0.9798, P0.0001; DPN group Area 0.5733, std.0.1211, 95% 0.3360to 0.8107, P0.5417; CI group Area 0.5, Std.0.1239, 95% 0.2572to 0.7428, P0.9999.
Detailed Description
The following examples are intended to illustrate the invention but are not intended to limit the scope of the invention. Unless otherwise specified, the technical means used in the examples are conventional means well known to those skilled in the art.
Example 1 screening for markers associated with anxiety and depression
For young men who face a greater psychopsychological stress in a long-term (7-month) task, peripheral venous blood is taken before and after the task is performed (5 ml of median venous blood of elbows of all subjects is obtained by a liberty military general hospital during 4-11 months in 2012, kept standing at room temperature for 30min, centrifuged at 1000 Xg for 15min, and frozen in a refrigerator at-70 ℃). The samples before the task were performed were 8, and the samples after the task were 12. 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. As a result, MMP-19 was found to be significantly elevated in post-task serum (p: 0.021458464) compared to pre-task serum (fig. 1), and MMP-19 was intended as a test index for indications of anxiety and depression.
Example 2 clinical validation of anxious depressed patients
1. Sampling
19 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 (experimental group), and 19 patients (control group) are collected in comparison with healthy volunteers who are detected by the first medical center of the general hospital of the liberation force at the same time and have normal indexes. Obtaining 5ml of median cubital venous blood of all subjects, standing for 30min at room temperature, centrifuging for 15min at 1000 Xg, subpackaging and freezing in a refrigerator at-70 ℃.
2. Procedure of experiment
The human MMP-19ELISA kit (purchased from CUSABIO under the code of CSB-EL014665HU) was used for the experiment. ELISA operation steps:
(1) the capture antibody was diluted in PBS to working concentration, added to ELISA plate wells (100 μ Ι/well), and incubated overnight at room temperature;
(2) adding 300 mul of washing buffer liquid for washing for 3 times;
(3) adding sample diluent, sealing at room temperature for 1h, wherein the volume of the sample diluent is 300 mu l/hole;
(4) adding 300 mul of washing buffer liquid for washing for 3 times;
(5) standard and sample: the standard was diluted with the sample diluent to a concentration of 8000pg/ml, 4000pg/ml, 2000pg/ml, 1000pg/ml, 500pg/ml, 250pg/ml, and serum samples at a concentration of 1: 100 dilution, adding ELISA plate holes (100 mu l/hole) respectively, directly adding sample diluent as blank control, covering with adhesive tape, and incubating at room temperature for 2 h;
(6) adding 300 mul of washing buffer liquid for washing for 3 times;
(7) adding detection antibody (100. mu.l/well) diluted to a proper concentration, and incubating at room temperature for 2 h;
(8) adding 300 mul of washing buffer liquid for washing for 3 times;
(9) adding Streptavidin-HRP (100. mu.l/well) diluted to a proper concentration, and incubating for 20min at room temperature in the dark;
(10) adding 300 mul of washing buffer liquid for washing for 3 times;
(11) adding chromogenic substrate (100 μ l/hole), and incubating at room temperature in dark for 20 min;
(12) stop solution (50. mu.l/well) was added, absorbance (OD) at a wavelength of 450nm was measured over 30 minutes, a standard curve was plotted (see Table 1), and MMP-19 concentration in serum was calculated.
TABLE 1 Standard Curve
| Standard article | 1 | 2 | 3 | 4 | 5 | 6 |
| Concentration (pg/ml) | 8000 | 4000 | 2000 | 1000 | 500 | 250 |
| OD mean value | 1.147 | 0.839 | 0.591 | 0.298 | 0.1 | 0.041 |
3. Results
ELISA results As shown in FIG. 2, serum MMP-19 levels were significantly higher in patients with depression-anxiety syndrome than in healthy volunteers, indicating that elevated serum MMP-19 levels are a marker for anxiety depression, p <0.0001(t test).
Example 3 differential diagnosis
1. Sampling
15 Diabetic Peripheral Neuropathy (DPN) and 15 Cerebral Infarction (CI) cases which are diagnosed in neurology of Beijing Kogyo hospital affiliated to capital medical university during 6 months 2020 to 10 months 2020 are collected for 10 cases in total by comparing with healthy volunteers which are from physical examination in Beijing Kogyo hospital affiliated to capital medical university at the same time and have normal indexes. Obtaining 5ml of median cubital venous blood of all subjects, standing for 30min at room temperature, centrifuging for 15min at 1000 Xg, subpackaging and freezing in a refrigerator at-70 ℃.
ELISA detection
Experimental procedures the procedure described in example 2 was referenced.
3. Results of the experiment
As shown in fig. 3, Diabetic Peripheral Neuropathy (DPN) showed no difference in serum MMP19 level (P ═ 0.5417) from normal human, and Cerebral Infarction (CI) showed no difference from control group (P ═ 0.9999).
The above results indicate that serum MMP-19 levels are not elevated in all neuropathies, and are specific for dysthymia.
Example 4 Receiver Operating Characteristic (ROC) Curve analysis
The subject performance characteristics of MMP-19 were analyzed using GraphPad prism8.3 software and ROC curves were plotted.
Determination of cut-off value: the mean value x and standard deviation s of the detection of the control healthy human serum were calculated, and then the CO value was determined according to the formula cut-off (CO) ═ x +3 s.
The specificity is true negative number/(true negative number + false positive number) × 100%
Sensitivity is 100% of true positive people/(true positive people + false negative people)
As shown in FIG. 4, the area under the curve (AUC) of ROC is: anxiety Depression (Depression), 0.867; diabetic Peripheral Neuropathy (DPN), 0.573; cerebral Infarction (CI), 0.5. At the optimal cutoff value (CO value: 298374.479pg/ml), the sensitivity and specificity of the marker was: specificity 96.6%, sensitivity: depression group: 58%, diabetic peripheral neuritis: 0% and cerebral infarction 0%.
These results indicate that MMP-19 can be used for clinical diagnosis of anxiety and depression.
Although the invention has been described in detail hereinabove with respect to a general description and specific embodiments thereof, it will be apparent to those skilled in the art that modifications or improvements may be made thereto based on the invention. Accordingly, such modifications and improvements are intended to be within the scope of the invention as claimed.
Claims (10)
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| CN113215249A (en) * | 2021-06-29 | 2021-08-06 | 中国人民解放军军事科学院军事医学研究院 | IL-26 for the identification and diagnosis of anxious depression |
| CN113252910A (en) * | 2021-06-29 | 2021-08-13 | 中国人民解放军军事科学院军事医学研究院 | Application of TIMP3 in anxiety and depression diagnosis |
| CN113373214A (en) * | 2021-06-18 | 2021-09-10 | 中国人民解放军军事科学院军事医学研究院 | Use of CD30 in diagnosis of cranial nerve related diseases |
| CN114774534A (en) * | 2022-04-15 | 2022-07-22 | 苏州惟慎生物科技有限公司 | Application of MMP19 as a biomarker related to early renal injury in early diagnosis of renal injury |
| CN116925231A (en) * | 2023-09-11 | 2023-10-24 | 中国人民解放军军事科学院军事医学研究院 | An antibody against MMP19 protein and its application |
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| CN114774534A (en) * | 2022-04-15 | 2022-07-22 | 苏州惟慎生物科技有限公司 | Application of MMP19 as a biomarker related to early renal injury in early diagnosis of renal injury |
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| CN116925231A (en) * | 2023-09-11 | 2023-10-24 | 中国人民解放军军事科学院军事医学研究院 | An antibody against MMP19 protein and its application |
| CN116925231B (en) * | 2023-09-11 | 2023-12-12 | 中国人民解放军军事科学院军事医学研究院 | An antibody against MMP19 protein and its application |
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