Biomarker for treating post-stroke depression and application thereof

Abstract

The invention discloses a biomarker for treating post-stroke depression and application thereof, and belongs to the technical field of biomedicine. The biomarker is a micro ribonucleic acid-146 (miR-146) family member, and specifically comprises miR146b (micro ribonucleic acid)-3p, miR146a (micro ribonucleic acid)-5p, miR146a (micro ribonucleic acid)-3p and miR146b (micro ribonucleic acid)-5p. Experiments prove that the expression of miR146b-3p and miR146a-5p in hippocampal tissues of mice in a PSD state is up-regulated, the expression of miR146a-3p and miR146b-5p is down-regulated, and the expression of the subtypes can be regulated to a normal level by using radix bupleuri liver soothing powder (CSS). The biomarker can be used for PSD diagnosis, illness state evaluation and treatment effect monitoring, provides a target spot for screening and research and development of PSD treatment drugs, and has important clinical application value and scientific research significance.

Description

Technical Field

[0001] The present invention discloses a biomarker for treating post-stroke depression and its application, belonging to the field of biomedical technology. Background Art

[0002] Post-stroke depression (PSD) is a common mood disorder among stroke survivors, characterized by persistent low mood, loss of interest, etc., which seriously hinders the recovery of the patient's motor and cognitive functions, increases the risk of stroke recurrence and mortality. The global incidence rate of PSD is 25% - 79%, and the combined incidence rate is 29% - 31%, bringing a huge burden to the patient's family and society.

[0003] The pathogenesis of PSD is complex, involving neurobiological changes, psychological factors and other aspects. At present, clinical treatment mostly uses antidepressant drugs such as fluoxetine, but there are problems such as limited efficacy and obvious side effects. The traditional Chinese medicine compound Chaihu Shugan San (CSS) has been proven to have a good therapeutic effect on PSD, but its mechanism of action has not been fully elucidated.

[0004] MicroRNA (miRNA) plays an important role in gene expression regulation and participates in multiple physiological processes such as neuroinflammation and synaptic plasticity. As an important carrier of intercellular communication, exosomes can carry miRNA to transmit biological information between cells and are closely related to the pathophysiological process of PSD. Therefore, finding specific biomarkers related to PSD is of great significance for the early diagnosis of the disease, monitoring of treatment effects and new drug research and development. Summary of the Invention

[0005] Aiming at the deficiencies of the prior art, the purpose of the present invention can be achieved through the following technical solutions: I. Determination of Biomarkers The present invention has confirmed through in vitro and in vivo experiments that four subtypes of the miR-146 family (miR146b-3p, miR146a-5p, miR146a-3p and miR146b-5p) can be used as biomarkers for treating PSD.

[0006] In in vitro experiments, an oxygen-glucose deprivation / reoxygenation (OGD / R) BV2 microglial cell model was constructed, and it was found that exosomes secreted by microglia in the OGD / R state could be taken up by hippocampal neurons and were related to the transmission of miR-146. In in vivo experiments, a PSD mouse model was constructed by combining photothrombotic stroke (PT) with chronic unpredictable mild stress (CUMS). Through RT-qPCR detection, it was found that: compared with healthy mice, the expressions of miR146b-3p and miR146a-5p in the hippocampal tissue of PSD mice were up-regulated, and the expressions of miR146a-3p and miR146b-5p were down-regulated; after treatment with Chaihu Shugan San, the expressions of each subtype were restored to normal levels.

[0007] II. Application of Biomarkers Diagnosis of PSD: By detecting the expression levels of miR146b-3p, miR146a-5p, miR146a-3p, and miR146b-5p in the hippocampus or blood of a subject, it can be determined whether the subject has PSD or assess the severity of the condition. If the test results show upregulation of miR146b-3p and miR146a-5p and downregulation of miR146a-3p and miR146b-5p, it suggests that the subject may have PSD or that the condition is in an active phase.

[0008] Screening for drugs to treat PSD: Using the expression levels of the four subtypes of the miR-146 family as indicators, potential drugs that can regulate their expression to normal levels can be screened. The specific screening steps include constructing a PSD model, drug intervention, and detecting miRNA expression levels. If a drug can reverse the abnormal expression of miR-146 subtypes in the PSD model, it is considered a potential therapeutic drug.

[0009] Monitoring the efficacy of PSD treatment: For PSD patients receiving treatment, the expression changes of miR146b-3p, miR146a-5p, miR146a-3p, and miR146b-5p in the body were dynamically monitored. If the expression of each subtype gradually returned to normal after treatment, it indicated that the treatment regimen was effective; if the expression did not improve significantly or remained abnormal, it suggested that the treatment regimen needed to be adjusted.

[0010] III. Drug Screening Kit This invention also provides a drug screening kit for treating PSD, comprising reagents such as specific primers, probes, or antibodies for detecting the expression levels of miR146b-3p, miR146a-5p, miR146a-3p, and miR146b-5p. This kit can rapidly and accurately detect the expression levels of target miRNAs in a model, providing a tool for high-throughput screening of drugs for treating PSD.

[0011] IV. Application of Bupleurum Liver-Soothing Powder Experiments have confirmed that Chaihu Shugan Powder can effectively regulate the expression of four subtypes of the miR-146 family in a dose-dependent manner. Among them, the high-dose (4.2 g / kg / day) Chaihu Shugan Powder showed the best regulatory effect, significantly improving depressive symptoms in PSD mice, reducing neuroinflammation, and promoting angiogenesis and synaptic function recovery. Therefore, Chaihu Shugan Powder can be used to prepare drugs that regulate the expression of the above biomarkers for the treatment of PSD.

[0012] The beneficial effects of this invention are: 1. This invention is the first to demonstrate that four subtypes of the miR-146 family exhibit specific abnormal expression in PSD (upregulation of miR146b-3p and miR146a-5p, and downregulation of miR146a-3p and miR146b-5p), and that this abnormality can be reversed by PSD treatment drugs (such as CSS). Figure 1-8 The experimental data validated the specificity of the biomarker from multiple dimensions, including in vitro cell models, in vivo animal models, behavioral, pathological, and molecular biology, providing an objective indicator for PSD diagnosis.

[0013] 2. Diagnostic methods developed based on this biomarker can achieve early diagnosis of PSD; drug screening kits can quickly screen potential therapeutic drugs and shorten the research and development cycle; efficacy monitoring methods can dynamically evaluate treatment effects and provide a basis for adjusting individualized treatment plans, solving the pain points of difficult diagnosis, slow drug screening, and subjective efficacy evaluation in the current diagnosis and treatment of PSD.

[0014] 3. Experiments have confirmed that CSS can treat PSD by regulating the expression of miR-146 subtypes, thereby inhibiting neuroinflammation, promoting angiogenesis, and improving synaptic function. Moreover, the high dose (4.2g / kg / day) is more effective than fluoxetine, providing clear dosage reference and mechanism support for the clinical application of CSS, and helping to promote the standardized application of traditional Chinese medicine compound in the treatment of PSD. Attached Figure Description

[0015] To more clearly illustrate the technical solutions in the embodiments of this disclosure or the prior art, the accompanying drawings used in the description of the embodiments or the prior art will be briefly introduced below. Obviously, those skilled in the art can obtain other drawings based on these drawings without creative effort.

[0016] Figure 1 The following is a diagram of the network pharmacology analysis results. a) CSS components screened by LC-MS; b) CSS drug-target network; c) Protein-protein interaction (PPI) network of CSS components and PSD-related proteins; d) Top ten core target genes in the PPI network; e) KEGG pathway enrichment analysis results; f) GO enrichment analysis results (biological process BP, cellular composition CC, molecular function MF).

[0017] Figure 2 In vitro experiments. (a) Morphology of microglia under normal culture conditions and after oxygen-glucose deprivation / reoxygenation / re-glucose (OGD / R) treatment; (b) Cell viability under different doses of serum containing ginseng and senna (CSS); Data are expressed as mean ± standard deviation. p<0.001, compared with the control group; ###p<0.001, compared with group A; ^^^p<0.001, compared with group B; +++p<0.001, compared with group C; &&&p<0.001, compared with group D, n=7; (c) Images taken by transmission electron microscopy (TEM); (d) Results of nanoparticle tracking analysis (NTA); (e) Results of flow cytometry detection of CD9, CD63 and CD81; (f) Fluorescent images of PKH26 labeling.

[0018] Figure 3 Relevant assessments of the primary sinus node disease (PSD) model: (a) a graph showing the percentage reduction in cerebral perfusion on the embolic side; (b) a graph showing the mean cerebral perfusion as measured by the PeriCam PSI system; (c) representative images of blood flow perfusion observed by the PeriCam PSI system in the control group; (d) a graph showing the difference in body weight between the day before the start of chronic unpredictable mild stress (CUMS) and the last day of drug treatment; (e) a graph showing the percentage of sucrose preference; (ab) data are expressed as mean ± standard deviation, ###p<0.001, compared with the control group; p < 0.001, compared with the model group, n = 3; (de) data are expressed as mean ± standard deviation, ### p < 0.001, compared with the control group; p < 0.001, n = 7 compared to the model group.

[0019] Figure 4 Behavioral assessment of mice with primary sinoatrial node disease (PSD); (a) Tail Suspension Test (TST): Graph of the percentage of time mice spent in a state of despair; (b) Forced Swimming Test (FST): Graph of representative movement trajectories; (c) Forced Swimming Test (FST): Graph of the percentage of time mice spent in a state of despair; (d) Open Field Test (OFT): Graph of total time spent at rest; (e) Open Field Test (OFT): Graph of distance traveled in the central region; (f) Open Field Test (OFT): Graph of distance traveled in the peripheral region. (g) Open field test (OFT): Representative motion trajectory records for each group; (h) Hematoxylin-eosin staining (HE): Representative images of hematoxylin-eosin staining; (i) Hematoxylin-eosin staining (HE): Statistical graph of semi-quantitative analysis of hematoxylin-eosin staining in the CA3 region of the hippocampus; (j) Hematoxylin-eosin staining (HE): Statistical graph of semi-quantitative analysis of hematoxylin-eosin staining in the cortical region; (a, cf) Data are expressed as mean ± standard deviation, p < 0.001, compared with the control group; p<0.001, compared with the model group, n=7; (ij) data are expressed as mean ± standard deviation, ###p<0.001, compared with the blank group; p<0.01, p<0.01, p < 0.001, n = 3 compared to the model group.

[0020] Figure 5 Immunofluorescence detection results of CD9 and CD63; (a) Representative immunofluorescence images of CD9 and CD63; (b) Statistical graph of CD9 fluorescence area; (c) Statistical graph of CD63 fluorescence area; Data are expressed as mean ± standard deviation, #p<0.05, ##p<0.01, compared with the blank group; p<0.05, p<0.01, p < 0.001, n = 3 compared to the model group.

[0021] Figure 6 Immunofluorescence detection results of CD81 and tumor susceptibility gene 101 (TSG101); (a) representative immunofluorescence images of CD81 and tumor susceptibility gene 101 (TSG101); (b) statistical graph of CD81 fluorescence area; (c) statistical graph of tumor susceptibility gene 101 (TSG101) fluorescence area; data are expressed as mean ± standard deviation, #p<0.05, ###p<0.001, compared with the control group; p<0.01, p < 0.001, n = 3 compared to the model group.

[0022] Figure 7(ab) Immunofluorescence assay results of calnexin and messenger RNA (mRNA) expression of exosome-associated marker proteins: (a) Representative immunofluorescence images of calnexin; (b) Statistical graph of calnexin fluorescence area; (cg) Quantitative analysis of messenger RNA (mRNA) expression, including (c) CD9, (d) CD63, (e) CD81, (f) tumor susceptibility gene 101 (TSG101), and (g) Alix protein; (hk) From reverse transcription to quantitative polymerization Statistical graph of relative expression levels of microRNA-146 obtained by enzyme chain reaction (RT-qPCR) experiments; (h) microRNA-146a-3p (miRNA-146a3p); (i) microRNA-146a-5p (miRNA-146a5p); (j) microRNA-146b-3p (miRNA-146b3p); (k) microRNA-146b-5p (miRNA-146b5p); Data are expressed as mean ± standard deviation, #p<0.05, ###p<0.001, compared with the control group; p<0.05, p<0.01, p < 0.001, n = 3 compared to the model group.

[0023] Figure 8 Quantitative analysis of mRNA expression levels including (a) S100A8, (b) interleukin-1β (IL-1β), (c) interleukin-6 (IL-6), (d) interleukin-10 (IL-10), (e) tumor necrosis factor-α (TNF-α), (f) vascular endothelial growth factor C (VEGFC), (g) vascular endothelial growth factor receptor 3 (VEGFR3), (h) synaptotropic protein 1, (i) synaptotropic protein 1, and (j) synaptotropic protein 14; data are expressed as mean ± standard deviation, #p<0.05, ##p<0.01, ###p<0.001, compared with the control group; p<0.05, p<0.01, p < 0.001, n = 3 compared to the model group. Detailed Implementation

[0024] The technical solutions of the embodiments of the present invention will be clearly and completely described below with reference to the accompanying drawings. Obviously, the described embodiments are only some embodiments of the present invention, and not all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of the present invention.

[0025] I. Experimental Materials laboratory animals BALB / c mice: male, 5 weeks old, weighing 18-22g, purchased from Beijing Vital River Laboratory Animal Technology Co., Ltd., license number: SCXK (Guangzhou) 2022-0063.

[0026] Sprague-Dawley (SD) rats: male, 7 weeks old, weighing 220-250g, purchased from the same company, used to prepare CSS-containing serum.

[0027] Rearing conditions: Specific pathogen-free (SPF) environment, constant temperature 25℃, light / dark cycle 12h / 12h, free access to food and water, and experiments began after 1 week of acclimatization.

[0028] Drugs and reagents Chaihu Shugan San (CSS): Composed of Bupleurum, Tangerine Peel, Ligusticum chuanxiong, Cyperus rotundus, Bitter Orange Peel, White Peony Root, and Licorice Root in a mass ratio of 4:4:3:3:3:3:1. The raw materials were purchased from China Kangmei Pharmaceutical Co., Ltd. The medicinal materials were mixed with distilled water at a ratio of 1:10 (mass / volume), refluxed and boiled for 2 hours, repeated twice, concentrated by rotary evaporation, and stored at -80℃ for later use.

[0029] Fluoxetine hydrochloride: Sigma-Aldrich, USA, catalog number #PHR1394, used as a positive control.

[0030] Key reagents: Bengal rose sodium salt (Coollab., China, catalog number #CR9581), thiazolyl blue (MTT, BioFroxx, Germany, catalog number #1334MG250), PBS buffer (Beyotime, China, catalog number #BL601A), HE staining kit (Beyotime, China, catalog number #C0105), RT-qPCR related reagents (miRNA first-strand cDNA synthesis kit, Novizan, Nanjing, catalog number #MR201; ChamQ Universal SYBR qPCR Master Mix, Novizan, Nanjing, catalog number #Q711).

[0031] Cells and Instruments Cells: BV2 microglia and HT22 hippocampal neurons were purchased from the Cell Bank of the Chinese Academy of Sciences (Kunming) and cultured in DMEM high glucose medium containing 10% fetal bovine serum (FBS) and 1% penicillin-streptomycin at 37°C in a 5% CO2 incubator.

[0032] Instruments: Liquid Chromatography-Mass Spectrometry (LC-MS, Waters Corporation ACQUITY I-Class system), Transmission Electron Microscope (TEM, Hitachi HT-7700), Nanoparticle Tracking Analyzer (NTA, ParticleMetrix ZetaView® PMX120), Real-time Quantitative PCR System (Bio-Rad Laboratories CFX96Touch), Behavioral Analysis System (Shanghai Xinruan, models XR-XQ202, XR-XQ203, XR-XZ301).

[0033] II. Experimental Methods Preparation of CSS-containing serum SD rats were randomly divided into a blank group and a CSS-containing serum group, with 10 rats in each group.

[0034] CSS-containing serum group: CSS decoction (4.2 g / kg) was administered by gavage twice daily for 7 consecutive days; control group: an equal volume of physiological saline was administered by gavage.

[0035] Two hours after the last administration, blood was collected from the abdominal aorta of rats, centrifuged at 3000×g for 15 min at 4℃, and repeated twice. The supernatant serum was collected, filtered through a 0.22μm filter, inactivated at 56℃ for 30 min, and then stored at -80℃ for later use.

[0036] In vitro experiments: Construction of OGD / R-BV2 cell model and detection of miR-146 Cell grouping: BV2 cells were seeded in 96-well plates (7000 cells / well), and after adhesion, they were divided into 10 groups (6 wells per group): Normal culture groups: control group (10% FBS), group A (7.5% FBS + 2.5% CSS-S), group B (5% FBS + 5% CSS-S), group C (2.5% FBS + 7.5% CSS-S), and group D (10% CSS-S).

[0037] OGD / R groups: Model group (OGD / R + 10% FBS), Group E (OGD / R + 7.5% FBS + 2.5% CSS-S), Group F (OGD / R + 5% FBS + 5% CSS-S), Group G (OGD / R + 2.5% FBS + 7.5% CSS-S), Group H (OGD / R + 10% CSS-S).

[0038] OGD / R treatment: OGD / R group cells were placed in sugar-free DMEM medium and incubated at 37°C for 3 hours in a 95% N2 + 5% CO2 incubator. Then, the medium was replaced with normal medium containing the corresponding concentration of CSS-S and cultured under normoxic conditions for 24 hours.

[0039] Cell viability assay (MTT assay): Add 5 mg / ml MTT reagent to each well, incubate at 37°C for 2 h, add 100 μl DMSO to dissolve formazan crystals, measure absorbance at 450 nm wavelength, and calculate cell viability.

[0040] Exosome extraction and identification: OGD / R-BV2 cell supernatant was collected, and cell debris was removed by centrifugation at 500×g for 10 min, 1000×g for 10 min, and 10000×g for 30 min. Exosome precipitate was then obtained by ultracentrifugation at 100000×g for 70 min. The precipitate was analyzed by TEM (excitosome analysis). Figure 2 c), NTA ( Figure 2 d) Flow cytometry Figure 2 e) Identify exosomes.

[0041] Exosome uptake assay: PKH26-stained exosomes were co-cultured with HT22 cells for 24 h, nuclei were stained with DAPI, and uptake was observed under a fluorescence microscope. Figure 2 f).

[0042] miR-146 detection: Total RNA was extracted from OGD / R-BV2 cells and exosomes, and the expression of miR146b-3p, miR146a-5p, miR146a-3p, and miR146b-5p was detected by RT-qPCR.

[0043] In vivo experiments: Construction and related tests of the PSD mouse model Model building: PT modeling: Mice were intravenously injected with 1% Bengal rose red (30mg / kg), and after anesthesia induced by 2% isoflurane, the left motor cortex was irradiated with a 534nm beam (60mW / cm²) for 20min to establish a focal thrombotic stroke model; 24h later, mice with neurological deficits (score >0) were screened using the Zea-Longa score.

[0044] CUMS modeling: Mice that successfully modeled PT were randomly divided into a model group, a fluoxetine group (10 mg / kg / day), a high-dose CSS group (4.2 g / kg / day), and a low-dose CSS group (2.1 g / kg / day), with 7 mice in each group; the blank group (7 mice) did not undergo PT modeling and were only fed normally.

[0045] CUMS stimulation: Mice were subjected to random stress (7 types of stress, including fasting for 24 hours, water deprivation for 24 hours, tilting the cage at 45° for 7 hours, etc., without repetition within 72 hours) for 5 consecutive weeks, while the mice were administered drugs by gavage daily. The model group was given an equal volume of water by gavage.

[0046] Behavioral tests: Weight measurement: Mice were weighed before and after CUMS, and the weight difference was calculated. Figure 3 d).

[0047] Sugar water preference test (SPT): During the training period (3 days), mice had free access to plain water and 1% sucrose solution. During the testing period, mice were housed individually, and their water intake was recorded over 24 hours. The sugar water preference rate was calculated. Figure 3 e).

[0048] Tail Suspension Test (TST): The mouse tail is suspended 10cm above the ground, and a 5-minute video is recorded. The duration of stillness in the last 4 minutes is analyzed. Figure 4 a).

[0049] Forced swimming test (FST): Mice were placed in a 30cm × 18cm cylindrical container (water temperature 25±2℃, water depth 15cm), and a 6-minute video was recorded. The last 4 minutes of immobility were analyzed. Figure 4 b, 4c).

[0050] Open field test (OFT): Mice were placed in the center of a 50cm×50cm×40cm open field, and a 5-minute video was recorded. The total immobility time and the distance moved in the central area were analyzed. Figure 4 d, 4e, 4g).

[0051] Pathological examination: Brain tissue was collected from mice after sacrifice, fixed in 4% formaldehyde for 7 days, embedded in paraffin and sectioned (5μm), stained with hematoxylin and eosin (HE), and the pathological changes in the hippocampus and cortex were observed. Figure 4 h), and performed semi-quantitative analysis ( Figure 4 i、4j).

[0052] Molecular biological detection: Immunofluorescence: After dewaxing and hydration of brain tissue sections, antigen retrieval and blocking were performed. Primary antibodies (CD9, CD63, CD81, TSG101, and calcium-linked protein) were added and incubated overnight at 4°C. Secondary fluorescent antibody was then incubated for 1 hour. Nuclei were stained with DAPI, and observation was performed under a fluorescence microscope. Figure 5 , Figure 6 , Figure 7 a).

[0053] RT-qPCR: Total RNA was extracted from hippocampal tissue, reverse transcribed to synthesize cDNA, and RT-qPCR was used to detect the miR-146 subtype ( Figure 7mRNA expression of hk), inflammatory factors (S100A8, IL1β, etc.), angiogenic factors (VEGFC, VEGFR3), and synapse-related proteins (Synphilin-1, etc.) Figure 8 ).

[0054] III. Experimental Results In vitro experimental results Cell viability: The survival rate of BV2 cells decreased significantly after OGD / R treatment (P<0.001), and increased after the addition of CSS-S. The 10% CSS-S group (H group) had the highest survival rate (47%), which was 9% higher than that of the model group (38%) (P<0.001). Figure 2 b).

[0055] Exosome identification: TEM showed that the exosomes were teacup-shaped. Figure 2 c), NTA showed particle sizes of 75-200 nm ( Figure 2 d), flow cytometry detected positive expression of CD9, CD63, and CD81 ( Figure 2 e) confirms successful exosome extraction.

[0056] Exosome uptake: PKH26-labeled red fluorescence was observed in HT22 cells, and the fluorescence increased with increasing exosome concentration. Figure 2 f), indicating that exosomes can be taken up by neurons.

[0057] miR-146 expression: In OGD / R-BV2 cells, the expression of miR146b-3p and miR146a-5p was upregulated, while the expression of miR146a-3p and miR146b-5p was downregulated (P<0.001 compared with the control group); after adding 10% CSS-S, the expression of each subtype returned to normal levels (P<0.001).

[0058] In vivo experimental results Behavioral studies: Compared with the model group, mice in the high-dose CSS group experienced reduced weight loss. Figure 3 d) Increased preference for sugary drinks ( Figure 3 e) The TST and FST stationary time is shortened ( Figure 4 a, 4c), OFT total stationary time decreased and central area movement distance increased ( Figure 4 The differences between d and 4e were statistically significant (P<0.001).

[0059] Pathology: In the model group mice, hippocampal neurons showed disordered arrangement and nuclear dissolution, and red necrotic neurons were visible in the cortex. Figure 4 h); Neuronal damage was significantly reduced in the high-dose CSS group, and the number of intact cells in the hippocampal CA3 region and cortex increased (h); Figure 4 i, 4j, P<0.001).

[0060] Immunofluorescence: In the model group, the expression of CD9, CD63, CD81, and TSG101 was decreased, while the expression of calcium-linked protein was increased in the hippocampus. Figure 5 , Figure 6 , Figure 7 a, 7b); the high-dose CSS group showed that the change was reversible (P<0.001).

[0061] RT-qPCR: miR146b-3p and miR146a-5p were upregulated and miR146a-3p and miR146b-5p were downregulated in the hippocampus of the model group. Figure 7 hk, P<0.001); the high-dose CSS group could restore the expression of all subtypes to normal (P<0.001); at the same time, the high-dose CSS group could reduce the mRNA level of inflammatory factors and restore the mRNA expression of VEGFC, VEGFR3 and synapse-related proteins (hk, P<0.001); Figure 8 (P<0.001) In the description of this specification, references to terms such as "an embodiment," "example," "specific example," etc., indicate that a specific feature, structure, material, or characteristic described in connection with that embodiment or example is included in at least one embodiment or example of the invention. In this specification, illustrative expressions of the above terms do not necessarily refer to the same embodiment or example. Furthermore, the specific features, structures, materials, or characteristics described may be combined in any suitable manner in one or more embodiments or examples.

[0062] The foregoing has shown and described the basic principles, main features, and advantages of the present invention. Those skilled in the art should understand that the present invention is not limited to the above embodiments. The embodiments and descriptions in the specification are merely illustrative of the principles of the invention. Various changes and modifications can be made to the invention without departing from its spirit and scope, and all such changes and modifications fall within the scope of the claimed invention.

Claims

1. A biomarker for the treatment of post-stroke depression, characterized in that, The biomarker is a micro ribonucleic acid-146 family member, including miR146b-3p, miR146a-5p, miR146a-3p and miR146b-5p.

2. The biomarker of claim 1, wherein, In the post-stroke depression state, the expression levels of miR146b-3p and miR146a-5p are up-regulated, and the expression levels of miR146a-3p and miR146b-5p are down-regulated; in the normal physiological state or after the post-stroke depression is effectively treated, the expression levels of each subtype return to the normal range.

3. Use of the biomarker of claim 1 or 2 in the preparation of a post-stroke depression diagnostic reagent.

4. Use according to claim 3, characterized in that, The diagnostic reagent determines whether a subject has post-stroke depression or assesses the severity of post-stroke depression by detecting the expression levels of miR146b-3p, miR146a-5p, miR146a-3p and miR146b-5p in the hippocampus tissue or blood of the subject.

5. Use of the biomarker of claim 1 or 2 in the screening of a post-stroke depression treatment drug.

6. Use according to claim 5, characterized in that, The use includes the following steps: (1) constructing a post-stroke depression cell model or animal model; (2) applying a drug to be screened to the model; (3) detecting the expression levels of miR146b-3p, miR146a-5p, miR146a-3p and miR146b-5p in the model; (4) if the drug to be screened can restore the abnormal expression of each miR-146 subtype in the model to the normal expression level, the drug is determined to be a potential post-stroke depression treatment drug.

7. Use of the biomarker of claim 1 or 2 in the preparation of a post-stroke depression treatment effect monitoring reagent.

8. Use according to claim 7, characterized in that, The monitoring reagent dynamically detects the expression level changes of miR146b-3p, miR146a-5p, miR146a-3p and miR146b-5p in a post-stroke depression patient receiving treatment, and evaluates the effectiveness of the treatment scheme.

9. A drug screening kit for treating post-stroke depression, characterized by, The kit contains reagents for detecting the expression levels of miR146b-3p, miR146a-5p, miR146a-3p and miR146b-5p, and the reagents include specific primers, probes or antibodies.

10. Use of Chaihu Shugan Powder in the preparation of a drug for regulating the expression of the biomarker of claim 1, and the use amount of the Chaihu Shugan Powder is 2.1 g / kg / day-4.2 g / kg / day.

Images