A vascular inflammation-related protein marker for assisting diagnosis or detection of cerebral hemorrhage and its application

Abstract

The invention discloses a vascular inflammation-related protein marker for auxiliary diagnosis or detection of cerebral hemorrhage and an application thereof. The protein marker is characterized in that the protein marker is a combination of KIM1 protein, IL6 protein, PTX3 protein and Gal-9 protein or KIM1 protein. The prepared kit for auxiliary diagnosis or detection of cerebral hemorrhage comprises RT-qPCR quantitative amplification primers of KIM1 transcriptome, IL6 transcriptome and PTX3 transcriptome. The advantage is that the detection kit based on detecting the transcriptome / protein expression level of KIM1 marker, IL6, PTX3, KIM1, Gal-9 combination marker and IL6, PTX3, and KIM1 combination marker can conveniently and quickly realize the detection of cerebral hemorrhage at the molecular level, and has high detection efficiency, high sensitivity and strong pertinence.

Description

Technical Field

[0001] The present invention relates to the technical field of auxiliary diagnosis of cerebral hemorrhage, and in particular to a vascular inflammation-related protein marker for auxiliary diagnosis or detection of cerebral hemorrhage and an application thereof. Background Art

[0002] Cerebral hemorrhage (abbreviated as ICH) is also called cerebral hemorrhage. It has various causes, most of which are caused by rupture of blood vessels due to hypertensive arteriosclerosis, so some people also call it hypertensive cerebral hemorrhage. Cerebral hemorrhage is a common acute cerebrovascular disease in middle-aged and elderly people, with high mortality and disability rates.

[0003] Brain damage after intracerebral hemorrhage includes primary damage caused by physical trauma and blood infiltration into the brain parenchyma to form hematomas that compress surrounding tissues, as well as secondary brain damage such as cerebral edema, blood-brain barrier destruction, inflammation, and impaired neuronal conduction. During secondary brain damage after intracerebral hemorrhage, the body's defense response relies heavily on inflammation. This involves the activation and polarization of glial cells (such as microglia or macrophages), the infiltration of peripheral inflammatory cells, and the release of proinflammatory factors. The activation of microglia / macrophages leads to polarization and subsequent release of inflammatory mediators such as interleukin 1β (IL-1β), tumor necrosis factor α (TNF-α), etc. These inflammatory mediators lead to neuronal apoptosis and blood-brain barrier destruction, causing severe brain damage.

[0004] Clinically, the diagnosis of cerebral hemorrhage mainly relies on imaging examinations. Currently, the most commonly used examination methods include head CT scans, head MRI scans, cerebral angiography, etc. These technologies can only be completed if the patient is sent to the hospital for treatment in time after the onset of the disease. Therefore, we urgently need a technology that can diagnose and predict cerebral hemorrhage at an early stage.

[0005] The detection of protein biomarkers plays a vital role in disease prediction, screening, treatment and long-term outcome assessment. Existing studies have identified many plasma predictors of ICH, such as interleukin-6 (IL-6), pentraxin-related protein 3 (PTX3), galectin-9 (Gal-9), etc. IL6 is a proinflammatory cytokine that has been widely studied as an indicator of systemic inflammation in acute brain injury. IL6 levels are associated with poor neurological function and large amounts of bleeding after ICH. PTX3 is an acute phase response protein that plays a key role in the inflammatory process and immune system regulation and is rapidly produced after infection, injury or inflammatory stimulation. Studies have found that elevated PTX3 levels in patients in the early stages of ICH are associated with adverse outcomes of ICH, including higher mortality and disability rates. Gal-9 is a glycoprotein belonging to the galectin family that has been shown to play a key regulatory role in inflammation and immune responses. Current studies have shown that in rat models of cerebral hemorrhage, Gal-9 levels are elevated, accompanied by an increase in M2 microglia and related anti-inflammatory factors. The determination of the levels of these protein factors has certain clinical value in the diagnosis and treatment of cerebral hemorrhage. Hepatitis A virus cellular receptor 1 (KIM1) is an immunoglobulin superfamily protein that is usually increased in the proximal tubules of damaged or diseased kidneys. Most studies on KIM1 have focused on kidney injury, emphasizing its high expression in renal tubular epithelial cells. However, studies on KIM1 in the nervous system are relatively limited, probably because its expression level in brain tissue is low and therefore has not been widely explored. Summary of the invention

[0006] The technical problem to be solved by the present invention is to provide a vascular inflammation-related protein marker and its application for auxiliary diagnosis or detection of cerebral hemorrhage with high detection efficiency, high sensitivity and strong specificity.

[0007] The technical solution adopted by the present invention to solve the above technical problem is: a vascular inflammation-related protein marker for auxiliary diagnosis or detection of cerebral hemorrhage, wherein the protein marker is KIM1 protein.

[0008] Furthermore, the above-mentioned vascular inflammation-related protein marker is used in the preparation of a kit for auxiliary diagnosis or detection of cerebral hemorrhage, wherein the kit includes RT-qPCR quantitative amplification primers for the KIM1 transcriptome, the nucleotide sequence of the forward amplification primer of the KIM1 transcriptome is shown in SEQ ID NO.1: 5′-GAATCTATGCTGGAGTCTGT-3′, and the nucleotide sequence of the reverse amplification primer of the KIM1 transcriptome is shown in SEQ ID NO.2: 5′-GTCTGATGTGCTGATGTCT-3′.

[0009] The present invention also provides a vascular inflammation-related protein marker for auxiliary diagnosis or detection of cerebral hemorrhage, wherein the protein marker is a combination of KIM1 protein, IL6 protein, PTX3 protein and Gal-9 protein.

[0010] The present invention also provides a vascular inflammation-related protein marker for auxiliary diagnosis or detection of cerebral hemorrhage, wherein the protein marker is a combination of KIM1 protein, IL6 protein and PTX3 protein.

[0011] Further, the above-mentioned vascular inflammation-related protein markers are used in the preparation of a kit for auxiliary diagnosis or detection of cerebral hemorrhage, wherein the kit includes RT-qPCR quantitative amplification primers for KIM1 transcriptome, IL6 transcriptome and PTX3 transcriptome, the nucleotide sequence of the forward amplification primer of the KIM1 transcriptome is shown in SEQ ID NO.1: 5′-GAATCTATGCTGGAGTCTGT-3′, and the nucleotide sequence of the reverse amplification primer of the KIM1 transcriptome is shown in SEQ ID NO.2: 5′-GTCTGATGTGCTGATGTCT-3′;

[0012] The nucleotide sequence of the forward amplification primer of the IL6 transcriptome is shown in SEQ ID NO.3: 5′-TAGAGTACCTCCAGAACAGATT-3′, and the nucleotide sequence of the reverse amplification primer of the IL6 transcriptome is shown in SEQ ID NO.4: 5′-AATAGTGTCCTAACGCTCATAC-3′;

[0013] The nucleotide sequence of the forward amplification primer of the PTX3 transcriptome is shown in SEQ ID NO.5: 5′-GCATAATAGGAACACTTGAGAC-3′, and the nucleotide sequence of the reverse amplification primer of the PTX3 transcriptome is shown in SEQ ID NO.6: 5′-CTGACAGAGACACAGCATT-3′.

[0014] Compared with the prior art, the advantages of the present invention are: the present invention is a vascular inflammation-related protein marker for auxiliary diagnosis of cerebral hemorrhage and its application, and discloses IL6, PTX3, KIM1, Gal-9 and IL6, PTX3, KIM1 and Gal-9 combined markers for auxiliary diagnosis of cerebral hemorrhage for the first time, and the transcriptome and protein levels of IL6, PTX3, KIM1 and Gal-9 are expressed in the blood of patients with cerebral hemorrhage. The mechanism may be that these vascular inflammation-related proteins continue to be highly expressed after inflammatory damage to the blood vessels, and participate in the pathological development process of cerebral vascular rupture and brain damage by increasing the secretion level of inflammatory factors. Therefore, the detection kit based on the detection of KIM1 marker, IL6, PTX3, KIM1, Gal-9 combined marker and IL6, PTX3, and KIM1 combined marker transcriptome / protein expression level can conveniently and quickly realize the detection of cerebral hemorrhage at the molecular level, with high detection efficiency, high sensitivity, strong pertinence, and is conducive to the early detection and timely treatment of patients with cerebral hemorrhage. BRIEF DESCRIPTION OF THE DRAWINGS

[0015] Figure 1 The differential expression diagrams of all vascular inflammation-related biomarkers between the cerebral hemorrhage group and the control group, where A is the volcano diagram of 76 inflammation-related biomarkers, and B is the expression level difference diagram of 18 inflammation-related biomarkers, *P<0.05, **P<0.01, ***P<0.001;

[0016] Figure 2 GO and KEGG enrichment analysis diagrams were performed for differentially expressed vascular inflammation-related proteins, where A is the top 20 GO enrichments based on the background of all annotated proteins, and B is the top 20 KEGG enrichment pathways based on the background of all annotated proteins;

[0017] FIG3 is a heat map showing the correlation between differentially expressed vascular inflammation-related proteins in patients with cerebral hemorrhage;

[0018] Figure 4 is a heat map of the correlation between differentially expressed proteins and clinical characteristics, where A is the correlation diagram between 18 inflammatory proteins and clinical characteristics in all groups, B is the correlation diagram between 18 inflammatory proteins and clinical characteristics in the cerebral hemorrhage group, and C is the correlation diagram between 18 inflammatory proteins and clinical characteristics in the control group, *P<0.05, **P<0.01;

[0019] Figure 5 The ELISA test results of IL6 protein, KIM1 protein, PTX3 protein and Gal-9 protein in the plasma of the control group and the cerebral hemorrhage group, where A is IL6, B is KIM1, C is PTX3, D is Gal-9, *P<0.05, **P<0.01, ***P<0.001;

[0020] Figure 6 The ROC curve analysis diagram of ELISA validation of the early diagnostic value of IL6 protein, PTX3 protein, KIM1 protein and Gal-9 protein and their combination for cerebral hemorrhage;

[0021] Figure 7 The relative expression results of RT-qPCR of KIM1, IL6 and PTX3 transcriptome in the control group and blood leukocytes; A is KIM1, B is IL6, C is PTX3, ****P<0.0001;

[0022] Figure 8 ROC curve diagram for RT-qPCR validation of IL6, PTX3, and KIM1 transcriptome expression levels in blood leukocytes. DETAILED DESCRIPTION

[0023] The present invention is further described in detail below with reference to the accompanying drawings.

[0024] Specific example 1: Screening of vascular inflammation-related proteins in cerebral hemorrhage.

[0025] 1. Clinical data of volunteers in the screening phase: This study collected volunteers from the Department of Neurosurgery, the First Affiliated Hospital of Ningbo University, including 26 patients with cerebral hemorrhage diagnosed by cerebral angiography (53.04±10.79, 13 males), and 26 control groups (53.08±6.67, 13 males) who were clinically confirmed to have no cerebrovascular lesions. The case group and the control group were strictly matched one to one in terms of gender and age, and fasting venous blood samples were collected from all participants for the detection of general biochemical indicators such as blood lipids and blood sugar and the detection of nucleic acid content in the blood. Their clinical data including gender, age, history of smoking and drinking, hypertension, and the results of routine blood biochemical tests were recorded. The clinical data of the first batch of cerebral hemorrhage cases and control groups used for screening differential proteins are shown in Table 1 below.

[0026] 2. Plasma extraction and biochemical analysis: During the clinical sample collection process of this study, about 6 mL of blood was drawn from the patient within 6 hours after admission. After centrifugation at 3200 rpm for 15 minutes at 4°C, the blood was separated and the upper plasma layer and the middle buffy coat layer were extracted. Blood glucose, total cholesterol, triglycerides, high-density lipoprotein, and low-density lipoprotein were quantitatively analyzed using an automatic biochemical analyzer (Olympus AU2700, Japan). The concentrations of apolipoprotein A, apolipoprotein B, and apolipoprotein E were measured using turbidimetric immunoassay technology, and the results are shown in Table 1.

[0027] Table 1. Comparison of clinical data between the cerebral hemorrhage case group and the control group in the first screening phase

[0028]

[0029] As shown in Table 1, the blood glucose level of patients with cerebral hemorrhage was significantly higher than that of the control group (P=0.018). There were no statistical differences in age, gender distribution, triglycerides, total cholesterol, high-density lipoprotein, low-density lipoprotein, apolipoprotein A, apolipoprotein B and apolipoprotein E between the two groups (P>0.05).

[0030] 3. Analysis of vascular inflammation-related proteins in cerebral hemorrhage by ultrasensitive multiplex targeted protein detection analysis (abbreviated as Olink): According to the manufacturer's guidelines, plasma samples from 26 patients with cerebral hemorrhage and 26 control patients were analyzed using the Olink® target 76 CVD II panel (Olink Proteomics AB, Sweden). This technology involves highly specific binding of target proteins to antibody probes labeled with dual oligonucleotides, followed by detection and quantification using a microfluidic real-time PCR instrument (Biomark HD, USA). The final detection readout is displayed as a standardized protein expression value, which is used for various biological analyses after log2 transformation.

[0031] Based on the Olink CVD II panel analysis, we found that 18 inflammation-related proteins were differentially expressed between the two groups of patients. Figure 1 As shown in center A, 14 proteins were upregulated in the ICH group, including IL6, PTX3, KIM1, Gal-9, and interleukin-4 receptor alpha (IL-4RA). In addition, B-type natriuretic peptide (BNP), TNF receptor superfamily member 10A (TNFRSF10A), heat shock protein 27 (HSP27), interleukin-1 receptor antagonist protein (IL-1ra), tumor necrosis factor receptor family member 10B (TRAIL-R2), carcinoembryonic antigen-related cell adhesion molecule 8 (CEACAM8), angiotensin-converting enzyme 2 (ACE2), growth hormone (GH), and P-selectin glycoprotein ligand-1 (PSGL-1) were all upregulated. In contrast, in the ICH group, four proteins were downregulated: CDO brother homolog (BOC), lipoprotein lipase (LPL), chymotrypsin-C (CTRC), and stem cell factor (SCF). Figure 1 The bar graph in center B further illustrates the fold change differences in protein expression. Table 2 shows the detailed information of the 18 differentially expressed proteins between ICH cases and controls.

[0032] Table 2 shows the detailed information of vascular inflammatory proteins that changed significantly between the ICH group and the control group

[0033]

[0034] Note: The fold change (FC) between the ICH group and the control group was calculated as log2 value. The p value (P<0.05) was calculated using t-test, indicating that there was a significant difference in the protein.

[0035] 4. Enrichment analysis and correlation analysis of differentially expressed proteins: When analyzing the differentially expressed protein (DEP) set between the ICH group and the control group, we used R software (Version 4.1.3) using the R package “Olink® Analyze” to determine the significant differentially expressed proteins. We used the ggplot2 package in R for data visualization and analysis, generating heat maps and volcano maps to visualize the data. In addition, we used the versatile data visualization package ggplot2 in R to perform gene ontology (GO) enrichment analysis and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis. In addition, in order to study the relationship between the protein expression patterns of the two groups, Spearman correlation analysis was performed. We constructed a protein-protein interaction (PPI) network for the differentially expressed proteins. Subsequently, we used igraph [1.4.1] and ggraph [2.1.0] to visualize these networks. Finally, the ROC curve was generated using the ROCR package, and two or more indicators were combined using Logistic regression analysis. The area under the curve (AUC) value of the combined diagnosis was displayed in the legend in the lower right corner to evaluate the classification performance of the data.

[0036] Functional enrichment analysis was used to elucidate the potential functions of differentially expressed proteins in plasma of ICH and control groups. Figure 2 As shown in Figure A, GO analysis showed that the differentially expressed proteins were mainly localized in the extracellular space, involved in the binding of TNF-related apoptosis-inducing ligand (TRAIL) to its receptor, regulating cell population proliferation, positively regulating T helper 2 (Th2) cell differentiation, and acting as virus receptors. Figure 2 As shown in B, the results of KEGG analysis showed that the differentially expressed inflammation-related protein factors were mainly concentrated in cytokine-cytokine receptor interactions, hematopoietic cell lineage, measles, PI3K-AKT and JAK-STAT signaling pathways.

[0037] The correlations among 18 inflammation-related proteins that were significantly different between the case and control groups, such as Figure 3As shown, BOC and IL6 showed the most significant negative correlation. At the same time, CTRC and PTX3, CTRC and CEACAM8, CTRC and HSP27, ACE2 and LPL, ACE2 and SCF, SCF and BNP, SCF and IL-1ra, IL-1ra and LPL all showed negative correlations. The other protein factors all showed positive correlations, and TRAIL-R2 and TNFRSF10A showed the most significant positive correlation. The correlations between these different factors may be closely related to the occurrence or poor prognosis of cerebral hemorrhage.

[0038] 5. Correlation between different inflammation-related proteins and clinical characteristics: We studied the association between 18 inflammation-related proteins and clinical characteristics of patients in each cohort. As shown in Figure 4A, the age of all participants was negatively correlated with PTX3 (P<0.05), gender was positively correlated with LPL and GH, and negatively correlated with ACE2 (P<0.05). It is worth noting that, Figure 4 As shown in Figures B and C, in the ICH group, PTX3 was negatively correlated with age and positively correlated with ApoE (P<0.05), but there was no significant correlation in the control group (P>0.05); similarly, Gal-9 was closely correlated with triglycerides, total cholesterol, low-density lipoprotein, and apolipoprotein B in the control group (P<0.05, Figure 4 C), but there was no significant correlation in the ICH group (P < 0.01; Figure 4 Middle B).

[0039] In summary, we collected blood from 26 pairs of patients with clinical cerebral hemorrhage and a control group, and used Olink ultra-sensitive multiple targeted protein detection and analysis technology to detect 76 vascular inflammation-related proteins in the patients' plasma. A total of 18 inflammation-related proteins were found to be differentially expressed between the two groups of patients, namely IL6, PTX3, KIM1, Gal-9, BOC, LPL, IL-4RA, BNP, TNFRSF10A, CTRC, HSP 27, SCF, IL-1ra, TRAIL-R2, CEACAM8, ACE2, GH, and PSGL-1.

[0040] Specific Example 2: Verify the diagnostic value of the vascular inflammation-related proteins screened in Specific Example 1 for cerebral hemorrhage.

[0041] The second batch of validation set clinical case group and control group samples were collected, with a total of 26 pairs of gender-age matched cerebral hemorrhage patients and non-cerebrovascular patients hospitalized in neurosurgery as the control group. The four proteins IL6, PTX3, KIM1, and Gal-9 with the most significant protein expression differences between the case group and the control group screened in Specific Example 1 were taken as the detection target proteins, and the difference in protein expression in patient plasma was verified by enzyme-linked immunosorbent assay (ELISA).

[0042] Table 3 is a comparison of clinical data between the cerebral hemorrhage case group and the control group in the second validation phase

[0043]

[0044] As shown in Table 3, the plasma glucose content in the cerebral hemorrhage group was significantly higher than that in the control group (P = 0.021). Figure 5 As shown in Figures A and B, the results of ELISA analysis showed that the levels of free IL6 and KIM1 proteins in the plasma of patients with cerebral hemorrhage were significantly higher than those in the control group (P<0.001). Figure 5 As shown in Figures C and D, the levels of PTX3 (P<0.01) and Gal-9 protein (P<0.05) in the plasma of patients with cerebral hemorrhage were both elevated and statistically significant. Subsequently, ROC analysis was performed to evaluate the diagnostic potential of each protein at different concentrations in the plasma of patients with cerebral hemorrhage. Figure 6 As shown in the figure, the area under the curve (AUC) of IL6 was 0.765, the 95% confidence interval was 0.621-0.909, the sensitivity was 76.92%, and the specificity was 80.77%. The AUC of PTX3 was 0.735, the 95% confidence interval was 0.591-0.879, the sensitivity was 61.54%, and the specificity was 84.62%. Among them, the AUC of KIM1 was the highest, 0.766, and the 95% confidence interval was 0.591-0.879. The diagnostic value was higher than other proteins, with a sensitivity of 76.92% and a specificity of 73.07%. The AUC of Gal-9 was 0.704, the 95% confidence interval was 0.560-0.849, the sensitivity was 65.39%, and the specificity was 80.77%. Interestingly, combining all four proteins in a multi-marker ROC analysis and subsequent logistic regression model significantly improved the diagnostic value AUC = 0.941, 95% confidence interval 0.885-0.997, and this combined approach also produced excellent sensitivity 76.93% and specificity 96.15%. These results highlight the potential of KIM1 as a diagnostic biomarker for ICH and also demonstrate that diagnostic performance can be improved through multi-marker combination.

[0045] Specific Example 3: Expand the real-time quantitative polymerase chain reaction RT-qPCR verification experiment of the case and control groups.

[0046] According to the ELISA test results in Specific Example 2, three inflammatory proteins (IL6, PTX3 and KIM1) with the highest ROC prediction values ​​were selected and verified at the mRNA level using RT-qPCR. Total RNA was isolated from blood buffy coat samples using TRIzol reagent. Subsequently, cDNA was synthesized from the RNA template using a cDNA reverse transcription kit (TransGen Biotech, China). RT-qPCR was performed on a Roche LightCycler 480 system using a SYBR Green SuperMix kit (TransGen Biotech, China). RT-qPCR primers were created using Primer6 software (Premier Biosoft, USA). The primer sequences are as follows:

[0047] The nucleotide sequence of the forward amplification primer of the KIM1 transcriptome is shown in SEQ ID NO.1: 5′-GAATCTATGCTGGAGTCTGT-3′, and the nucleotide sequence of the reverse amplification primer of the KIM1 transcriptome is shown in SEQ ID NO.2: 5′-GTCTGATGTGCTGATGTCT-3′; the nucleotide sequence of the forward amplification primer of the IL6 transcriptome is shown in SEQ ID NO.3: 5′-TAGAGTACCTCCAGAACAGATT-3′, and the nucleotide sequence of the reverse amplification primer of the IL6 transcriptome is shown in SEQ ID NO.4: 5′-AATAGTGTCCTAACGCTCATAC-3′; the nucleotide sequence of the forward amplification primer of the PTX3 transcriptome is shown in SEQ ID NO.5: 5′-GCATAATAGGAACACTTGAGAC-3′, and the nucleotide sequence of the reverse amplification primer of the PTX3 transcriptome is shown in SEQ ID NO.6: 5′-CTGACAGAGACACAGCATT-3′; the nucleotide sequence of the forward amplification primer of the internal reference gene ACTB is shown in SEQ ID NO. The nucleotide sequence of the reverse amplification primer of the internal reference gene ACTB is shown in SEQ ID NO.7: 5′-ATTGCCGACAGGATGCAGA-3′, and the nucleotide sequence of the reverse amplification primer of the internal reference gene ACTB is shown in SEQ ID NO.8: 5′-CAGGAGGAGCAATGATCTTGAT-3′; 2 −△△Ct The relative expression of mRNA was analyzed by the method.

[0048] The results are as follows Figure 7 As shown in Figure 2, the RT-qPCR results were consistent with the ELISA results. Figure 7As shown in A, the expression level of IL6 transcriptome in the buffy coat of patients with cerebral hemorrhage was significantly higher than that in the control group (P<0.001). Figure 7 PTX3 in B (P<0.0001) and Figure 7 The transcriptome expression level of KIM1 (P<0.0001) in C was significantly increased in the blood leukocytes of patients with cerebral hemorrhage. Figure 8 As shown, the results of ROC curve analysis showed that the IL6 transcriptome (AUC=1.00), PTX3 transcriptome (AUC=1.00) and KIM1 transcriptome (AUC=0.98) in the blood leukocytes of patients with cerebral hemorrhage all had very high diagnostic values.

[0049] The above description is not intended to limit the present invention, and the present invention is not limited to the above examples. Any changes, modifications, additions or substitutions made by a person skilled in the art within the spirit and scope of the present invention shall also fall within the protection scope of the present invention.

Claims

1. Use of a vascular inflammation-related protein marker in the preparation of a kit for auxiliary diagnosis or detection of cerebral hemorrhage, characterized in that: The protein marker is KIM1 protein.

2. Use of a vascular inflammation-related protein marker according to claim 1 in preparing a kit for auxiliary diagnosis or detection of cerebral hemorrhage, characterized in that: The kit comprises RT-qPCR quantitative amplification primers for the KIM1 transcriptome, the nucleotide sequence of the forward amplification primer for the KIM1 transcriptome is shown in SEQ ID NO.1: 5′-GAATCTATGCTGGAGTCTGT-3′, and the nucleotide sequence of the reverse amplification primer for the KIM1 transcriptome is shown in SEQ ID NO.2: 5′-GTCTGATGTGCTGATGTCT-3′.

3. A vascular inflammation-related protein marker for auxiliary diagnosis or detection of cerebral hemorrhage, characterized in that: The protein marker is a combination of KIM1 protein, IL6 protein, PTX3 protein and Gal-9 protein.

4. A vascular inflammation-related protein marker for auxiliary diagnosis or detection of cerebral hemorrhage, characterized in that: The protein marker is a combination of KIM1 protein, IL6 protein and PTX3 protein.

5. Use of the vascular inflammation-related protein marker according to claim 4 in preparing a kit for auxiliary diagnosis or detection of cerebral hemorrhage, characterized in that: The kit includes RT-qPCR quantitative amplification primers for KIM1 transcriptome, IL6 transcriptome and PTX3 transcriptome, The nucleotide sequence of the forward amplification primer of the KIM1 transcriptome is shown in SEQ ID NO.1: 5′-GAATCTATGCTGGAGTCTGT-3′, and the nucleotide sequence of the reverse amplification primer of the KIM1 transcriptome is shown in SEQ ID NO.2: 5′-GTCTGATGTGCTGATGTCT-3′; The nucleotide sequence of the forward amplification primer of the IL6 transcriptome is shown in SEQ ID NO.3: 5′-TAGAGTACCTCCAGAACAGATT-3′, and the nucleotide sequence of the reverse amplification primer of the IL6 transcriptome is shown in SEQ ID NO.4: 5′-AATAGTGTCCTAACGCTCATAC-3′; The nucleotide sequence of the forward amplification primer of the PTX3 transcriptome is shown in SEQ ID NO.5: 5′-GCATAATAGGAACACTTGAGAC-3′, and the nucleotide sequence of the reverse amplification primer of the PTX3 transcriptome is shown in SEQ ID NO.6: 5′-CTGACAGAGACACAGCATT-3′.

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