Application of fbl gene expression level detection agent in detection of postpartum depression and detection system
By combining FBL gene expression level detectors with other genes, a non-invasive quantitative detection system was developed, which solves the problem of difficulty in early diagnosis of postpartum depression in existing technologies and achieves high-precision detection of postpartum depression.
Patent Information
- Application Number
- CN202210959543.0
- Authority / Receiving Office
- CN · China
- Patent Type
- Patents(China)
- Current Assignee / Owner
- Filing Date
- 2021-04-02
- Publication Date
- 2025-11-04
- Estimated Expiration
- 2041-04-02
AI Technical Summary
Current technology lacks effective means and quantitative standards for early diagnosis of postpartum depression, leading to serious adverse consequences for mothers and infants.
By combining FBL gene expression level detectors with a combination of IL10, RAD51, UBA52, NHP2, RPL13A, HNRNPA2B1, and SPI1 genes, a non-invasive quantitative detection system was developed for the early diagnosis of postpartum depression.
It achieves high-precision early diagnosis of postpartum depression, with an area under the ROC curve exceeding 0.75, making it suitable as a clinical diagnostic reagent and capable of accurately determining the occurrence and development of postpartum depression.
Smart Images
Figure CN115261461B_ABST
Abstract
Description
[0001] The application is a divisional application of the invention patent with the title of a postpartum depression detection reagent, system and application, the application number of 202110360705.4, and the application date of April 2, 2021. TECHNICAL FIELD
[0002] The application belongs to the field of biochemical detection, and particularly relates to application of FBL gene expression level detection substance in postpartum depression detection and a detection system. BACKGROUND
[0003] Postpartum depression (PPD) affects 10% to 15% of postpartum women and is one of the most prominent factors affecting postpartum women's mood disorders. Its clinical symptoms usually occur within two weeks after delivery, and patients have persistent and severe emotions, including negative emotions (such as sadness, anxiety, worthlessness or despair), low energy and social isolation tendency. Postpartum depression can have short-term and long-term negative effects on the development, cognition, emotion and behavior of newborns. Studies have shown that maternal depression is a risk factor for premature infants and low birth weight infants, infant malnutrition and developmental delay, and it has gradually become a major public health problem.
[0004] Postpartum depression is the result of the combined effects of multiple factors. Recent studies have proposed hypotheses about the physiological and pathological mechanisms of postpartum depression, including abnormal changes in hormones, neurotransmitters and inflammatory factors, as well as genetics, social environment, psychological state, infant delivery method, etc. However, there is no conclusive evidence to support any of the above conclusions, and the physiological and pathological mechanisms of postpartum depression remain largely unclear. Due to the complexity of the pathogenesis of postpartum depression, there is currently a lack of effective means and quantitative standards to diagnose postpartum depression, resulting in the fact that postpartum depression can often only be judged after the fact, leading to serious adverse consequences for infants and mothers.
[0005] Early diagnosis will be of great significance to the prevention and treatment of postpartum depression and the avoidance of serious adverse consequences caused by postpartum depression. SUMMARY
[0006] In view of the above defects or improvement needs of the prior art, the application provides application of FBL gene expression level detection substance in postpartum depression detection and a detection system, which aims to help early diagnosis and diagnosis of postpartum depression through non-invasive quantitative detection, strengthen intervention on postpartum depression, and thus avoid the serious adverse effects of postpartum depression on mothers and infants.
[0007] To achieve the above-mentioned purpose, the application proposes the following solutions:
[0008] Application of FBL gene expression level detection substance in preparation of postpartum depression detection product.
[0009] wherein the sequence of the primer is
[0010] forward primer: 5'-TCTGGCAACCAGGTATTCAAGGC-3',
[0011] reverse primer: 5'-CATCGGCATTGCAACGTTTGGACT-3'.
[0012] wherein the gene detection chip is GSE45603 chip.
[0013] The application of the combination of the FBL gene expression level detection substance and the gene expression level detection substance of at least one of IL10, RAD51, UBA52, NHP2 and RPL13A in preparing a postpartum depression detection product.
[0014] The postpartum depression detection system comprises
[0015] The gene expression level acquisition module is configured to acquire the expression level parameter of the FBL gene and provide the parameter to the judgment module.
[0016] The judgment module is configured to take the FBL gene expression level parameter as the input of a linear classifier and judge whether the sample is from a postpartum depression patient.
[0017] Further, the postpartum depression detection reagent comprises a combination of one or more of the HNRNPA2B1, IL10, RAD51, UBA52, NHP2, RPL13A, FBL and SPI1 gene expression level detection reagents.
[0018] Preferably, the postpartum depression detection reagent comprises a combination of one or more of the HNRNPA2B1, FBL and SPI1 gene expression level detection reagents.
[0019] Preferably, the postpartum depression detection reagent comprises the HNRNPA2B1, FBL and SPI1 gene expression level detection reagents.
[0020] Preferably, the postpartum depression detection reagent comprises the gene mRNA expression level detection reagent.
[0021] Preferably, the postpartum depression detection reagent comprises the GSE45603 chip.
[0022] Preferably, the postpartum depression detection reagent comprises the HNRNPA2B1, FBL and / or SPI1 gene methylation level detection reagent.
[0023] According to another aspect of the present application, a postpartum depression detection system is provided, comprising a gene expression level acquisition module and a judgment module.
[0024] The gene expression level acquisition module is configured to acquire the expression levels of one or more of the genes HNRNPA2B1, IL10, RAD51, UBA52, NHP2, RPL13A, FBL, and SPI1, and provide the acquired expression levels to the judgment module.
[0025] The judgment module takes the expression levels of one or more of the genes HNRNPA2B1, IL10, RAD51, UBA52, NHP2, RPL13A, FBL, and SPI1 as inputs of a classifier, and judges whether the sample is from a postpartum depression patient.
[0026] Preferably, the postpartum depression detection system has the gene expression level acquisition module configured to acquire the expression levels of one or more of the genes HNRNPA2B1, FBL, and SPI1.
[0027] Preferably, the postpartum depression detection system has the gene expression level acquisition module configured to acquire the expression levels of the genes HNRNPA2B1, FBL, and SPI1.
[0028] The combination of one or more of the reagents for detecting the expression levels of the genes HNRNPA2B1, IL10, RAD51, UBA52, NHP2, RPL13A, FBL, and SPI1 is used to prepare a non-invasive quantitative detection reagent for postpartum depression.
[0029] Overall, the above technical solutions conceived by the present application can achieve the following beneficial effects compared with the prior art:
[0030] The postpartum depression detection reagent provided by the present application is applied to a postpartum depression detection system, and the area under the curve of the Receiver Operating Characteristic curve (ROC) of the subjects in the test system for the genes HNRNPA2B1, IL10, RAD51, UBA52, NHP2, RPL13A, FBL, and SPI1 all exceeds 0.75, when the area under the curve reaches 0.7 to 0.9, it is precision in clinical detection, and when it exceeds 0.9, it is high precision, and is very suitable as an early diagnosis reagent for postpartum depression.
[0031] In a preferred embodiment, the non-invasive quantitative detection can be performed in high throughput by the GSE45603 chip, so as to accurately perform quantitative detection of postpartum depression. In a preferred embodiment, the occurrence and development of postpartum depression can be more accurately judged by detecting the methylation levels of the target genes. Attached Figure Description
[0032] Figure 1 This is a graph showing the results of gene association network analysis related to postpartum depression in Example 2. Figure 1 In the middle, A represents the first gene interaction module network; Figure 1 In the middle B, there is a second gene interaction module network. Figure 1 C in the middle represents the third gene interaction module network;
[0033] Figure 2 Example 3 uses a postpartum depression detection system with different gene expression levels, and the area under the ROC curve is shown.
[0034] Figure 3 This is the result of the mouse modeling test for postpartum depression in Example 4;
[0035] Figure 4 This is the result of gene expression detection in mice with postpartum depression and normal controls in Example 4. Detailed Implementation
[0036] To make the objectives, technical solutions, and advantages of this invention clearer, the invention will be further described in detail below with reference to embodiments. It should be understood that the specific embodiments described herein are merely illustrative and not intended to limit the invention. Furthermore, the technical features involved in the various embodiments of this invention described below can be combined with each other as long as they do not conflict with each other.
[0037] The postpartum depression detection reagent provided by this invention includes one or more combinations of HNRNPA2B1, IL10, RAD51, UBA52, NHP2, RPL13A, FBL, and SPI1 gene expression level detection reagents.
[0038] Among them, HNRNPA2B1 gene is heterogeneous nuclear ribonucleoprotein A2 / B1 gene; IL10 gene is interleukin-10 gene; RAD51 gene is recombinase RAD51 gene; UBA52 gene is ubiquitin-ribosome fusion protein 52 gene; NHP2 is ribonucleoprotein 2 gene; RPL13A gene is ribosomal protein L13a housekeeping gene; FBL gene is fibrin gene; and SPI1 gene is Spi-1 proto-oncogene.
[0039] The statistics show that the mRNA expression difference of the above genes in the postpartum depression patients and the normal samples is more than twice obvious and the significance P value is less than 0.05; HNRNPA2B1, IL10 and RAD51 genes are in the same center position of the postpartum depression related gene correlation network, and can affect the same metabolic pathway, and the influence of HNRNPA2B1 is the most obvious; UBA52, NHP2, RPL13A, FBL are in the same center position of the postpartum depression related gene correlation network, and can affect the same metabolic pathway, and the influence of FBL is the most obvious; SPI1 gene is in the center position of another postpartum depression related gene correlation network. Therefore, the joint research of HNRNPA2B1, FBL and SPI1 shows that the expression level of mRAN in the postpartum depression patients and the normal samples is very clear.
[0040] The detection system using the mRNA expression of the three genes as the detection index is used to detect 21 blood samples (16 samples of postpartum depression patients and 5 normal control samples), and the area under the ROC curve is above 0.75, and the highest reaches 0.902, and since the three genes are in different modules, the correlation between them is not strong, and the joint index of the three can more accurately judge the postpartum depression sample.
[0041] The following is an example:
[0042] Example 1 Statistical analysis of expression difference of normal control and postpartum depression patients
[0043] Data source: The gene differential expression data of postpartum depression patients and normal controls is from the GEO database (Gene Expression Omnibus) GSE45603 data set, which includes 5 series of normal controls and 16 postpartum depression patients whole blood samples, and the samples are analyzed by GPL10558 platform (Illumina HumanHT-12V4.0 expression beadchip).
[0044] The R software (version number is 3.6.2) is used to analyze the expression difference of HNRNPA2B1, IL10, RAD51, UBA52, NHP2, RPL13A, FBL and SPI1 in normal control and postpartum depression patient samples. The results show that the expression difference of the above genes in the postpartum depression patients is obvious, and the P value is less than 0.05 and (|expression difference|) is greater than or equal to 0.2.
[0045] The expression difference and P value of the target genes HNRNPA2B1, IL10, RAD51, UBA52, NHP2, RPL13A, FBL and SPI1 used in the application are as follows:
[0046]
[0047] The above data show that the expression of the above genes in the mRNA of the postpartum depression sample and the normal sample shows significant difference.
[0048] Example 2 Postpartum depression related gene association network analysis
[0049] For the postpartum depression differentially expressed genes including HNRNPA2B1, IL10, RAD51, UBA52, NHP2, RPL13A, FBL, SPI1 genes, the GO database is used for automatic gene function annotation, the DAVID database is used for function and metabolic pathway analysis, the key molecular functions, biological processes, cell components and metabolic pathways of the above genes are visualized, and the interaction relationship between the above genes and diseases and drugs is analyzed by KEGG. As shown in the results Figure 1 The above genes are all enriched in the top 5 metabolic pathways of the differentially expressed genes in the postpartum depression and normal control in the GO analysis and KEGG analysis, and the statistical significance P value is <0.01. It shows that the above genes play a key role in participating in the metabolic pathways of postpartum depression.
[0050] In order to improve the accuracy, verify the correctness of HNRNPA2B1, IL10, RAD51, UBA52, NHP2, RPL13A, FBL, SPI1 genes as postpartum depression biochemical detection markers, GSEA analysis (GSEA version 4.0.3, http: / / software.broadinstitute.org / gsea / index.jsp) is applied to gene enrichment analysis, and the P value of the above genes is also <0.05. It is verified that the above genes participate in the metabolic pathways related to postpartum depression, and can be used as screening markers for postpartum depression. And it shows that the high expression of HNRNPA2B1 gene is related to endocytosis function, the low expression of FBL gene is related to long-range inhibition of brain, and the low expression of SPI1 gene is related to cell proliferation.
[0051] The Cytoscape software was used to analyze the gene interaction of the differentially expressed genes in the normal control and postpartum depression patients. The interactions with a reliability score greater than 0.4 were selected to analyze the relationship between the differentially expressed genes. A total of 673 differentially expressed genes were introduced into the Cytoscape software for analysis. The CytoHubba plug-in in the Cytoscape software was used to retrieve the hub gene. The hub gene was screened according to the degree algorithm, and the top 30 genes were defined as the hub gene. The results showed that the HNRNPA2B1, IL10, RAD51, UBA52, NHP2, RPL13A, FBL, and SPI1 genes were ranked within the top 30 hub genes, and the hub gene scores were as follows:
[0052]
[0053] Example 3: Postpartum depression detection system
[0054] A postpartum depression detection system includes a gene expression level acquisition module and a judgment module.
[0055] The gene expression level acquisition module is used to acquire the expression level of one or more genes of HNRNPA2B1, IL10, RAD51, UBA52, NHP2, RPL13A, FBL, and SPI1 genes, and provide the judgment module.
[0056] The judgment module takes the expression levels of HNRNPA2B1, IL10, RAD51, UBA52, NHP2, RPL13A, FBL, and SPI1 genes as the input of the linear classifier to determine whether the sample is from a postpartum depression patient.
[0057] The GEO database (Gene Expression Omnibus) GSE45603 dataset was used for testing, and the ROC curve was drawn as shown in Figure 2 The AUC is shown in the following table:
[0058]
[0059] The above results confirm that the above genes can be used as postpartum depression markers, and the detection system can effectively distinguish postpartum depression samples from normal samples, reaching a medium effective level, and being suitable as a clinical screening indicator.
[0060] Example 4: Animal experiment verification
[0061] The present study adopts chronic pre-pregnancy stress to induce the construction of postpartum depression mouse model, and evaluates the expression of HnRNPA2B1, Fibrillarin (FBL) and Spi-1 proto-oncogene (SPI1) genes in the whole blood of female mice. 7-week-old Balb / c female mice are randomly divided into two groups, the blank control group is not given any stimulation, and the control group of mice is given chronic stress for 3 weeks. After the stress, the blank control group and the stress group are randomly taken to mate with male mice to get pregnant, and after delivery, two groups are formed: the blank control group and the stress combined with delivery group, i.e. the postpartum depression model group. Three weeks after delivery, the behavior of mice in each group is detected, including forced swimming test, sugar water preference test, and tail suspension test, to evaluate whether the modeling is successful. Three weeks after delivery, the whole blood of female mice in each group is taken, and the expression of HnRNPA2B1, Fibrillarin (FBL) and Spi-1 proto-oncogene (SPI1) genes is detected by real-time fluorescent quantitative PCR.
[0062] The results of behavior detection: after behavior detection, it is found that the model is successfully established, and the results are shown in Figure 3
[0063] The primers used in the gene expression test are as follows:
[0064]
[0065]
[0066] Gene expression: the mRNA expression of three genes in the whole blood is detected by qRT-PCR, and the detection results are shown in Figure 4
[0067] The results show that the above genes in the postpartum depression model mice have obvious expression difference compared with the normal control, which confirms that the gene expression level detection reagent of the above genes is suitable as the postpartum depression detection reagent.
[0068] Those skilled in the art will readily understand that the above description is only the preferred embodiment of the present application, and is not intended to limit the present application. Any modification, equivalent replacement and improvement within the spirit and principle of the present application should be included in the protection scope of the present application.
Claims
1. Application of FBL gene expression level detection material in the preparation of postpartum depression detection products.
2. The application according to claim 1, wherein, The gene expression level detection material is a gene mRNA expression level detection material, which includes primers or gene detection chips.
3. The application according to claim 2, wherein, The sequence of the primer is as follows: Forward primer: 5'-TCTGGCAACCAGGTATTCAAGGC-3', Reverse primer: 5'-CATCGGCATTGCAACGTTTGGACT-3'.
4. The application according to claim 2, wherein, The gene detection chip is a GSE45603 chip.
5. Application of the combination of FBL gene expression level detection substance with at least one of IL10, RAD51, UBA52, NHP2, and RPL13A gene expression level detection substances in the preparation of postpartum depression detection products.
6. The application according to claim 5, wherein, The gene expression level detection material is a gene mRNA expression level detection material.
7. A postpartum depression detection system, characterized in that, include The gene expression level acquisition module is used to obtain the expression level parameters of the FBL gene in the sample and provide them to the judgment module; The judgment module uses the FBL gene expression level parameter as input to a linear classifier to determine whether the sample comes from a patient with postpartum depression.
8. The postpartum depression detection system according to claim 7, characterized in that, Gene expression level detection refers to the detection of gene mRNA expression levels.