Method for analyzing correlation between ANXA2 and chronic cerebral hypoperfusion blood-brain barrier damage index and inflammatory injury index

Abstract

The invention provides a correlation analysis method for ANXA2 and chronic cerebral hypoperfusion blood-brain barrier damage indexes and inflammatory injury indexes, and belongs to the field of data analysis. The invention aims to solve the problems of limitation and low authenticity due to the fact that mechanisms and influence factors of ANXA2 are mostly explored by constructing CCH animal models during correlation research of ANXA2 and chronic cerebral hypoperfusion. A CCH model is constructed through a patient suffering from carotid artery stenosis, two lesions MCI and WML caused by the CCH are taken as dependent variables, the ANXA2 level is taken as an independent variable, and the correlation between the ANXA2 level and MCI and WML is researched, so that the correlation between the ANXA2 level and brain barrier damage and inflammatory injury of the CCH is researched. Compared with a traditional CCH animal model, the clinical test flexibility is better, and the credibility is higher.

Description

Technical Field

[0001] The present invention relates to the technical field of data analysis, and in particular to a method for analyzing the correlation between ANXA2 and chronic cerebral hypoperfusion blood-brain barrier destruction indicators and inflammatory injury indicators. Background Art

[0002] Chronic cerebral hypoperfusion (CCH) refers to a pathological and physiological state in which the perfusion of brain tissue is lower than the physiological threshold for a long time, causing the brain tissue to be in a state of ischemia and hypoxia for a long time. Its imaging features are reduced cerebral blood flow leading to white matter lesions (WML). The mechanisms of CCH include oxidative stress, neuroinflammation, and abnormal function of the neurotransmitter system. The above pathological and physiological processes can also lead to damage of the blood-brain barrier (BBB). CCH can lead to progressive and persistent neurological and cognitive impairment (MCI). Compared with acute stroke, CCH has a longer intervention time window. Its early diagnosis is of great significance for the prevention, treatment and improvement of prognosis of MCI and WML. ANXA2 is a calcium-dependent phospholipid-binding protein that is mainly distributed on the cell membrane and participates in multiple biochemical processes, such as cell membrane transport, endocytosis, exocytosis, tissue remodeling, angiogenesis and immune regulation.

[0003] At present, many studies have explored the mechanisms of chronic cerebral hypoperfusion (CCH) leading to white matter lesions (WML) and mild cognitive impairment (MCI), including microglial activation, BBB destruction, myelin loss, neuroinflammatory response, etc. A review of the literature found that ANXA2 can affect BBB permeability and neuroinflammatory response in many neurological diseases, affecting the course and prognosis of the disease. However, most of the existing studies were conducted through animal models of CCH, and there is no research on the correlation between ANXA2 and blood-brain barrier destruction and inflammatory damage in CCH. For example, Li and Tian found that rats had obvious cognitive impairment, BBB ultrastructure destruction, Claudin-5 downregulation, and increased IL-1β expression due to CCH. Kenta Tezuka constructed a mouse model of multiple sclerosis and found that the expression of ANXA2 and a series of molecules involved in ANXA2 signaling were upregulated when the BBB was destroyed. Qin-YuHan et al. constructed a CCH mouse model by bilateral common carotid artery ligation and found that Claudin-5 expression was reduced under CCH conditions. EunChae Lee constructed a rat model of chronic cerebral hypoperfusion and used PCR to detect the transcription levels of Claudin-5, IL-6, and IL-1β in brain tissues. It was found that the transcription level of Claudin-5 was significantly reduced and the transcription level of IL-1β was significantly increased. Based on this, we established the following hypothesis: under CCH conditions, due to BBB destruction and neuroinflammatory damage, ANXA2 and IL-1β are upregulated, and Claudin-5 is reduced, leading to the occurrence of WML and MCI.

[0004] The animal model of CCH was chosen for research because CCH is generally in the late stage when discovered, so it is difficult to obtain a large number of early or initial CCH patients for correlation research, which has major limitations and affects the authenticity. Summary of the invention

[0005] The technical problems to be solved by the present invention are:

[0006] In order to solve the problem that most studies on the correlation between ANXA2 and chronic cerebral hypoperfusion explore its mechanism and influencing factors by constructing CCH animal models, which has limitations and affects the authenticity.

[0007] The present invention adopts the following technical solutions to solve the above technical problems:

[0008] The present invention provides a method for analyzing the correlation between ANXA2 and chronic cerebral hypoperfusion blood-brain barrier destruction index and inflammatory injury index, comprising the following steps:

[0009] S100. Construct a chronic cerebral hypoperfusion model, including dividing patients with carotid stenosis into a carotid stenosis group and a control group, and the carotid stenosis group was divided into a moderate stenosis group and a severe stenosis group according to the severity of carotid stenosis; the subjects in the carotid stenosis group were divided into an MCI group and a non-MCI group to analyze the correlation between MCI and peripheral blood ANXA2, Claudin-5 and IL-1β levels in chronic cerebral hypoperfusion; the subjects in the carotid stenosis group were divided into a mild WML group, a moderate WML group and a severe WML group to analyze the correlation between the degree of WML and peripheral blood ANXA2, Claudin-5, IL-1β levels and MCI in chronic cerebral hypoperfusion;

[0010] S200, correlation experiment division, including S210 correlation analysis of blood-brain barrier and inflammatory damage under different degrees of carotid artery stenosis, S220 correlation analysis of MCI with blood-brain barrier and neuroinflammation in chronic cerebral hypoperfusion, and S230 correlation analysis of WML with blood-brain barrier and neuroinflammation in chronic cerebral hypoperfusion.

[0011] Furthermore, step S210 performs correlation analysis on the blood-brain barrier and inflammatory damage under different degrees of carotid artery stenosis, including:

[0012] S211. Correlation analysis was performed on the peripheral blood ANXA2 levels, IL-1β levels and Claudin-5 levels in the moderate carotid stenosis group, severe carotid stenosis group and control group to prove whether the changes in ANXA2 levels, IL-1β levels and Claudin-5 levels in carotid stenosis and chronic cerebral hypoperfusion are the same, and further prove whether patients with carotid stenosis can replace patients with chronic cerebral hypoperfusion as a model of chronic cerebral hypoperfusion;

[0013] S212, after it is proved that the carotid stenosis patients can be used as chronic cerebral hypoperfusion models according to step S211, the MCI prevalence rates of the moderate carotid stenosis group, the severe carotid stenosis group and the control group are compared to prove whether the carotid stenosis is related to the prevalence of MCI, and further prove whether MCI can be used as an indicator for determining the chronic cerebral hypoperfusion model;

[0014] S213. Comparison of the prevalence of WML in the moderate carotid stenosis group, severe carotid stenosis group and control group was used to prove whether the chronic cerebral hypoperfusion model is related to the prevalence of WML, and further prove whether WML can be used as an indicator to judge the chronic cerebral hypoperfusion model.

[0015] Further, step S220 performs correlation analysis on MCI, blood-brain barrier and neuroinflammation during chronic cerebral hypoperfusion, including:

[0016] S221. Correlation analysis was performed on the peripheral blood ANXA2 level, Claudin-5 level and IL-1β level in the MCI group and the non-MCI group to prove whether MCI is correlated with ANXA2 level, Claudin-5 level and IL-1β level, and then to determine whether the chronic cerebral hypoperfusion model is correlated with ANXA2 level, Claudin-5 level and IL-1β level;

[0017] S222. When MCI is correlated with ANXA2 levels, Claudin-5 levels, and IL-1β levels, a multivariate logistic regression equation was constructed for the ANXA2 and IL-1β levels in peripheral blood serum. Multivariate binary logistic regression analysis was performed to determine whether ANXA2 and IL-1β were risk factors for MCI. Based on the analysis results, it was determined whether ANXA2 and IL-1β levels could be used to diagnose MCI.

[0018] Further, step S230 performs correlation analysis on WML, blood-brain barrier and neuroinflammation during chronic cerebral hypoperfusion, including:

[0019] S231. Correlation analysis was performed on the peripheral blood ANXA2 level, Claudin-5 level and IL-1β level in the mild WML group, moderate WML group and severe WML group to prove whether WML is correlated with ANXA2 level, Claudin-5 level and IL-1β level, and then to determine whether the chronic cerebral hypoperfusion model is correlated with ANXA2 level, Claudin-5 level and IL-1β level;

[0020] S232. When WML is associated with ANXA2, Claudin-5 and IL-1β levels, a multivariate logistic regression equation was constructed for ANXA2, IL-1β and Claudin-5 levels in peripheral blood serum. Multivariate binary logistic regression analysis was used to demonstrate whether ANXA2 and IL-1β were risk factors for WML. Based on the analysis results, it was demonstrated whether ANXA2, IL-1β and Claudin-5 levels could be used to diagnose WML.

[0021] Furthermore, in step S211, the correlation between the serum ANXA2, IL-1β and Claudin-5 levels in the carotid artery stenosis group and the control group was analyzed by rank sum test.

[0022] Furthermore, in step S212, the chi-square test is used to analyze whether the chronic cerebral hypoperfusion model is narrow and the correlation between the degree of narrowing and the prevalence of MCI.

[0023] Furthermore, in step S221, the correlation between MCI and ANXA2 level, IL-1β level and Claudin-5 level was analyzed by Wilcoxon Z rank sum test.

[0024] Furthermore, in step S231, the correlation between the severity of WML and the levels of ANXA2, Claudin-5 and IL-1β was analyzed by Kruskal-Wallis H rank sum test.

[0025] Furthermore, in step S100, the carotid artery stenosis degree of the moderate stenosis group is 50%-69%, and the carotid artery stenosis degree of the severe stenosis group is 70%-99%.

[0026] Compared with the prior art, the present invention has the following beneficial effects:

[0027] The present invention discloses a correlation analysis method between ANXA2 and blood-brain barrier destruction index and inflammatory injury index of chronic cerebral hypoperfusion. Patients with carotid artery stenosis greater than 50% are used as CCH models. Experiments prove that the two have similar changes in ANXA2 level, IL-1β level and Claudin-5 level in vivo. Patients with carotid artery stenosis can be used as CCH models. When performing correlation analysis between ANXA2 and blood-brain barrier destruction and inflammatory injury of CCH, two lesions MCI and WML caused by CCH are used as dependent variables, and ANXA2 level is used as an independent variable. By studying the correlation between ANXA2 level and CCH blood-brain barrier destruction and inflammatory injury, the correlation between ANXA2 level and CCH blood-brain barrier destruction and inflammatory injury is analyzed. The correlation between ANXA2 level and MCI and WML was studied, and then the correlation between ANXA2 level and brain barrier destruction and inflammatory damage in CCH was studied. This is because chronic cerebral hypoperfusion CCH leads to white matter lesions WML and mild cognitive impairment MCI, while brain protein lesions WML and mild cognitive impairment MCI cause BBB destruction and neurological verification damage; in this process, Claudin-5 levels decrease, IL-1β levels increase, and ANXA2 levels increase; compared with traditional CCH animal models, clinical trials have better flexibility, and the credibility of correlation research conclusions is higher and more objective. BRIEF DESCRIPTION OF THE DRAWINGS

[0028] Figure 1 This is a diagram of periventricular white matter cystic changes in an embodiment of the present invention;

[0029] Figure 2 This is a deep brain white matter lesion diagram in an embodiment of the present invention;

[0030] Figure 3 This is a curve diagram of predicting MCI through the ROC curve of ANXA2 and IL-1β in an embodiment of the present invention;

[0031] Figure 4This is a curve diagram of predicting WML through the ROC curve of ANXA2, IL-1β and Claudin-5 in the embodiment of the present invention. DETAILED DESCRIPTION

[0032] In order to make the above-mentioned objects, features and advantages of the present invention more obvious and easy to understand, specific embodiments of the present invention are described in detail below with reference to the accompanying drawings.

[0033] Specific implementation plan 1: Combine Figures 1 to 4 As shown, the present invention provides a correlation analysis method between ANXA2 and chronic cerebral hypoperfusion blood-brain barrier destruction index and inflammatory injury index, comprising the following steps:

[0034] S100. Sample selection and sample division. A certain number of elderly patients aged 55 to 80 years old were randomly selected. All the patients were required to complete cranial MR examination and carotid color Doppler ultrasound examination. The patients were divided into a carotid stenosis group and a control group. The carotid stenosis group was further divided into a moderate stenosis group (50%-69%) and a severe stenosis group (70%-99%) according to the severity of carotid stenosis. The subjects in the carotid stenosis group were divided into an MCI group and a non-MCI group to study the correlation between MCI and peripheral blood ANXA2, Claudin-5 and IL-1β levels in chronic cerebral hypoperfusion. The subjects in the carotid stenosis group were divided into a mild WML group, a moderate WML group and a severe WML group to study the correlation between the degree of WML and peripheral blood ANXA2, Claudin-5, IL-1β levels and MCI in chronic cerebral hypoperfusion.

[0035] S200, peripheral blood sample collection, fasting venous blood was collected from all research subjects, each sample was no less than 9 ml, and was naturally allowed to stand at room temperature for 30 minutes, 4°C, 3000 rpm, and centrifuged for 20 minutes to collect the supernatant, which was packaged into EP tubes, marked with groups and names, and stored in a -80°C refrigerator. If precipitation occurred during the storage process, it needed to be centrifuged again; during the experiment, the double antibody sandwich enzyme-linked immunosorbent assay was used for operation;

[0036] S300, correlation experiment division, including:

[0037] S310. Correlation analysis of blood-brain barrier and inflammatory damage under different degrees of carotid artery stenosis, including:

[0038] S311. Correlation analysis was performed on the levels of ANXA2, IL-1β and Claudin-5 in peripheral blood in the moderate carotid stenosis group, severe carotid stenosis group and control group to prove whether the changes of ANXA2, IL-1β and Claudin-5 levels in carotid stenosis and chronic cerebral hypoperfusion were the same;

[0039] S312. Comparison of the prevalence of MCI in the moderate carotid stenosis group, severe carotid stenosis group and control group to prove whether carotid stenosis is associated with the prevalence of MCI;

[0040] S313. Comparison of the prevalence of WML in the moderate carotid stenosis group, severe carotid stenosis group and control group was used to prove whether carotid stenosis is associated with the prevalence of WML.

[0041] S320. Correlation analysis between MCI and blood-brain barrier and neuroinflammation in chronic cerebral hypoperfusion, including:

[0042] S321. Correlation analysis was performed on peripheral blood ANXA2 levels, Claudin-5 levels and IL-1β levels in the MCI group and the non-MCI group to prove whether MCI is correlated with ANXA2 levels, Claudin-5 levels and IL-1β levels;

[0043] S322. When MCI is associated with ANXA2, Claudin-5 and IL-1β levels, a multivariate logistic regression equation was constructed for the ANXA2 and IL-1β levels in peripheral blood serum. Multivariate binary logistic regression analysis was performed to prove whether ANXA2 and IL-1β were risk factors for MCI. Based on the analysis results, it was proved whether ANXA2 and IL-1β levels could be used to diagnose MCI.

[0044] S330. Correlation analysis between WML and blood-brain barrier and neuroinflammation in chronic cerebral hypoperfusion, including:

[0045] S331. Correlation analysis was performed on peripheral blood ANXA2 levels, Claudin-5 levels and IL-1β levels in the mild WML group, moderate WML group and severe WML group to prove whether WML is correlated with ANXA2 levels, Claudin-5 levels and IL-1β levels;

[0046] S332. When WML is associated with ANXA2 levels, Claudin-5 levels, and IL-1β levels, a multivariate logistic regression equation was constructed for ANXA2 levels, IL-1β levels, and Claudin-5 levels in peripheral blood serum. Multivariate binary logistic regression analysis was used to demonstrate whether ANXA2 and IL-1β were risk factors for WML. Based on the analysis results, it was demonstrated whether ANXA2 levels, IL-1β levels, and Claudin-5 levels could be used to diagnose WML.

[0047] The other combinations and connection relationships of this embodiment are the same as those of the first embodiment.

[0048] experiment

[0049] Study subjects:

[0050] A total of 65 patients hospitalized in the Department of Neurology, the First Affiliated Hospital of Harbin Medical University from January 2023 to December 2023 were selected, including 49 males and 16 females. All enrolled patients were required to have complete clinical baseline data and complete cranial MRI examination and carotid artery color Doppler ultrasound examination.

[0051] Inclusion criteria:

[0052] Patients aged 55-80 years with complete clinical and statistical data, and all patients or their families signed informed consent.

[0053] Exclusion criteria:

[0054] (1) Patients with acute cerebral hemorrhage or a history of cerebral hemorrhage shown in brain CT. (2) Patients with congenital developmental abnormalities, vascular malformations or aneurysms. (3) Patients with severe hypotension, anemia, endocrine and hypoxic diseases. (4) Patients with a history of Alzheimer's disease, epilepsy or Parkinson's disease. (5) Patients with severe dysfunction of the heart, liver and kidney. (6) Patients with white matter lesions due to other reasons (genetic, immune, tumor and toxic). (7) Patients with contraindications to cranial MRI examination. (8) Pregnant women. (9) Patients with mental illness or other diseases who are unwilling to cooperate with the examination and are taking sedatives, anti-anxiety and anti-depressant drugs.

[0055] Grouping method:

[0056] The subjects were divided into groups according to the different research purposes: (1) divided into carotid stenosis group and control group. The carotid stenosis group was further divided into moderate stenosis group (50%-69%) and severe stenosis group (70%-99%) according to the severity of carotid stenosis. The correlation between carotid stenosis and peripheral blood ANXA2, Claudin-5, IL-1β levels, WML degree and MCI was studied. (2) The subjects with carotid stenosis >50% were divided into MCI group and non-MCI group to study the correlation between MCI and peripheral blood ANXA2, Claudin-5 and IL-1β levels during chronic cerebral hypoperfusion. (3) The subjects with carotid stenosis >50% were divided into mild WML group, moderate WML group and severe WML group to study the correlation between WML degree and peripheral blood ANXA2, Claudin-5, IL-1β levels and MCI during chronic cerebral hypoperfusion.

[0057] Data collection of enrolled patients

[0058] (1) General information collection: gender, age, medical history of hypertension, diabetes, hyperlipidemia, hyperhomocysteinemia, smoking and drinking, etc.

[0059] (2) Clinical data collection: diagnosis (whether it is cerebral infarction), complications (whether there is concurrent infection), and the presence or absence of mild cognitive impairment (Moca-B scale).

[0060] (3) Examination and test data collection: head MRI (Frzekas score), color Doppler ultrasound of neck vessels.

[0061] Among them, the scoring criteria for the Moca-B scale and frzekas are as follows:

[0062] (1) Montreal Cognitive Assessment-Basic (MoCA-B): The total score is 30 points. The cut-off value for identifying MCI is: ≤19 points for illiterate and primary school group; ≤22 points for secondary school group; ≤24 points for college group.

[0063] (2) Fazekas grading: Based on the FLAIR image score, score = A + B. 1-2 points for mild; 3-4 points for moderate; 5-6 points for severe. A. Periventricular white matter lesions (PVWM): 0 points for no lesions; 0 points for cap-shaped or pencil-like thin lesions ( Figure 1 a) is 1 point; the lesion is a smooth halo ( Figure 1 b) is 2 points; irregular periventricular high signal extending to the deep white matter ( Figure 1 c) is 3 points. B Deep white matter signal (DWM): No lesion is 0 points; punctate lesions ( Figure 2 a) is 1 point; the lesion begins to fuse ( Figure 2 b) is 2 points; large-area fusion of lesions ( Figure 2 c) is 3 points.

[0064] SPSS 26.0 software was used for statistical analysis. The measured data that conformed to the normal distribution were expressed as mean ± standard deviation. The data were expressed as P25, P75, and the inter-group comparison was performed by two independent sample T test (t), and the inter-group comparison was performed by F test (F); the measurement data that did not conform to the normal distribution were expressed as M (P25, P75), and the inter-group comparison was performed by Mann-Whitney U (Z) rank sum test, and the inter-group comparison was performed by Kruskal Wallis (H) rank sum test; the categorical variables were expressed as frequency (percentage, %), and the inter-group comparison was performed by chi-square test or Fisher's exact probability method; the measurement data that conformed to the linear trend and normal distribution, the correlation between samples was analyzed by Pearson correlation analysis. The measurement data or rank data that did not conform to the normal distribution, the correlation between samples was analyzed by Spearman correlation analysis. The variables with statistically significant inter-group differences (P < 0.05) were used to construct the Logistic regression model. The diagnostic statistical significance of each detection index was analyzed by ROC curve, and the optimal critical value of the biomarker was determined according to the Youden index, and the sensitivity and specificity were calculated. All the above test methods were considered statistically significant when the test level was P < 0.05.

[0065] Experimental results:

[0066] After peripheral blood sample collection and double antibody sandwich ELISA in step S200, the experimental results obtained are as follows:

[0067] (1) When analyzing the correlation between blood-brain barrier and inflammatory damage at different degrees of carotid artery stenosis, the comparative experiments and results are as follows:

[0068] ① The levels of ANXA2, IL-1β, and Claudin-5 in peripheral blood of the moderate carotid artery stenosis group, severe carotid artery stenosis group, and control group were compared. In order to determine whether there were differences in the levels of ANXA2, IL-1β, and Claudin-5 in serum between the carotid artery stenosis group and the control group, the rank sum test was used to compare the differences between the two groups.

[0069] The results are as follows: The levels of ANXA2, IL-1β, and Claudin-5 in the carotid stenosis group were higher than those in the control group, and the difference was highly statistically significant (P < 0.01). Spearman correlation was used to determine the correlation between ANXA2, IL-1β, and Claudin-5 and carotid stenosis. It was found that ANXA2, IL-1β, and Claudin-5 were positively correlated with the degree of carotid stenosis, and the difference was highly statistically significant (P < 0.01). See Tables 1 and 2 for details.

[0070] Table 1 Comparison of serum ANXA2, IL-1β and Claudin-5 levels among three groups

[0071]

[0072] Table 2 Correlation analysis between ANXA2, IL-1β and Claudin-5 and carotid artery stenosis

[0073]

[0074] ②Comparison of the prevalence of MCI in the moderate carotid stenosis group, severe carotid stenosis group and control group. In order to clarify whether there is a difference in the prevalence of MCI between the moderate carotid stenosis group, severe carotid stenosis group and control group, a chi-square test was performed. The results are as follows: the prevalence of MCI in the control group was 30%, the prevalence of MCI in the moderate carotid stenosis group was 58.8%, and the prevalence of MCI in the severe carotid stenosis group was 67.9%. The difference in the prevalence of the three groups was statistically significant (P < 0.05). See Table 3 for details.

[0075] Table 3 Comparison of MCI rates between the carotid stenosis group and the control group

[0076]

[0077] (2) When analyzing the correlation between MCI and blood-brain barrier and neuroinflammation during chronic cerebral hypoperfusion, the comparative experiments and results are as follows:

[0078] ① The peripheral blood ANXA2, Claudin-5, and IL-1β levels in the MCI group and the non-MCI group were compared. In order to clarify whether there were differences in the serum ANXA2, IL-1β, and Claudin-5 levels and the degree of MCI between the MCI group and the non-MCI group, the Wilcoxon (Z) rank sum test was used to compare the differences between the two groups. It was found that the peripheral blood serum ANXA2 and IL-1β levels in the MCI group were higher than those in the non-MCI group, and the difference was highly statistically significant (P < 0.01). Spearman correlation analysis was used to compare the correlation between the two groups, and the results showed that the ANXA2 and IL-1β levels were correlated with MCI, and the difference was highly statistically significant (P < 0.01). The correlation between the Claudin-5 level and MCI was not significant (P > 0.05). See Tables 4 and 5 for details.

[0079] Table 4 Comparison of peripheral serum ANXA2, Claudin-5, and IL-1β levels between the MCI group and the non-MCI group

[0080]

[0081] Table 5 Correlation analysis of peripheral serum ANXA2, IL-1β, and Claudin-5 levels in the MCI group and the non-MCI group

[0082]

[0083] ②Multivariate binary logistic regression analysis was performed on the influencing factors of MCI, and the ANXA2 and IL-1β levels in peripheral blood serum were included to construct a multivariate logistic regression equation. The results showed that increased ANXA2 and IL-1β levels were risk factors for MCI, and the differences were statistically significant (P < 0.05). See Table 6 for details.

[0084] Table 6 Multivariate binary logistic regression analysis of factors affecting MCI

[0085]

[0086] The MCI group was set as a positive event, and the non-MCI group was set as a negative event. Figure 3 As shown, the AUC of ANXA2 was 0.836 (95% CL: 0.720-0.952), which had diagnostic value, P < 0.01, and its Cut-off value was 3.365 ng / ml, with a sensitivity of 72.4% and a specificity of 93.7%; the AUC of IL-1β was 0.830 (95% CL: 0.706-0.953), which had diagnostic value, P < 0.01, and its Cut-off value was 17.202 pg / ml, with a sensitivity of 79.3% and a specificity of 75%.

[0087] (3) When studying the correlation between WML and blood-brain barrier and neuroinflammation in chronic cerebral hypoperfusion, the comparative experiments and results are as follows:

[0088] ① The levels of ANXA2, Claudin-5, and IL-1β in peripheral blood of mild WML group, moderate WML group, and severe WML group were compared. In order to clarify whether there were differences in the levels of ANXA2, IL-1β, and Claudin-5 in the serum of the three groups of patients, the KruskalWallis (H) rank sum test was used to compare the differences between the groups. The results were as follows: the higher the levels of ANXA2, Claudin-5, and IL-1β in peripheral blood, the more severe the degree of WML, and the difference was highly statistically significant (P < 0.01). Spearman correlation analysis was used to analyze the correlation between the three groups, and the results were as follows: the levels of ANXA2, Claudin-5, and IL-1β in peripheral blood were positively correlated with the severity of WML, and the difference was highly statistically significant (P < 0.01). See Tables 7 and 8 for details.

[0089] Table 7 Analysis of differences in peripheral blood ANXA2, Claudin-5, and IL-1β levels among the three groups

[0090]

[0091] Table 8 Correlation analysis of peripheral blood ANXA2, Claudin-5 and IL-1β levels in three groups

[0092]

[0093] ②Ordered Logistic regression analysis was performed on the influencing factors of WML, and the levels of ANXA2, Claudin-5, and IL-1β in peripheral blood were included to construct an ordered Logistic regression equation. The results showed that increased ANXA2, Claudin-5, and IL-1β were risk factors for WML, and the differences were statistically significant (P < 0.05). See Table 9 for details.

[0094] Table 9 Ordered Logistic Regression Analysis of WML Influencing Factors

[0095]

[0096] Mild WML was set as a negative event, and moderate to severe WML was set as a positive event. Figure 4 As shown, the AUC of ANXA2 was 0.745 (95% CL: 0.601-0.889), with a weak diagnostic value, P value = 0.011, its cut-off value was 3.619 ng / ml, sensitivity was 59.4%, and specificity was 92.3%. The curve of Claudin-5 had a P value of 0.106, P>0.05, and the difference was not statistically significant. The AUC of IL-1β was 0.760 (95% CL: 0.611-0.908), with a moderate diagnostic value, P value = 0.007, its cut-off value was 19.456 pg / ml, sensitivity was 65.6%, and specificity was 84.6%.

[0097] Although the present invention is disclosed as above, the protection scope of the present invention is not limited thereto. Those skilled in the art may make various changes and modifications without departing from the spirit and scope of the present invention, and these changes and modifications will fall within the protection scope of the present invention.

Claims

1. A method for analyzing the correlation between ANXA2 and blood-brain barrier destruction index and inflammatory injury index of chronic cerebral hypoperfusion, characterized in that: The following steps are involved: S100. Construct a chronic cerebral hypoperfusion model, including dividing patients with carotid stenosis into a carotid stenosis group and a control group, and the carotid stenosis group was divided into a moderate stenosis group and a severe stenosis group according to the severity of carotid stenosis; the subjects in the carotid stenosis group were divided into an MCI group and a non-MCI group to analyze the correlation between MCI and peripheral blood ANXA2, Claudin-5 and IL-1β levels in chronic cerebral hypoperfusion; the subjects in the carotid stenosis group were divided into a mild WML group, a moderate WML group and a severe WML group to analyze the correlation between the degree of WML and peripheral blood ANXA2, Claudin-5, IL-1β levels and MCI in chronic cerebral hypoperfusion; S200, correlation experiment division, including S210 correlation analysis of blood-brain barrier and inflammatory damage under different degrees of carotid artery stenosis, S220 correlation analysis of MCI with blood-brain barrier and neuroinflammation in chronic cerebral hypoperfusion, and S230 correlation analysis of WML with blood-brain barrier and neuroinflammation in chronic cerebral hypoperfusion.

2. The method for analyzing the correlation between ANXA2 and chronic cerebral hypoperfusion blood-brain barrier destruction index and inflammatory injury index according to claim 1, characterized in that: Step S210 performs correlation analysis on the blood-brain barrier and inflammatory damage under different degrees of carotid artery stenosis, including: S211. Correlation analysis was performed on the peripheral blood ANXA2 levels, IL-1β levels and Claudin-5 levels in the moderate carotid stenosis group, severe carotid stenosis group and control group to prove whether the changes in ANXA2 levels, IL-1β levels and Claudin-5 levels in carotid stenosis and chronic cerebral hypoperfusion are the same, and further prove whether patients with carotid stenosis can replace patients with chronic cerebral hypoperfusion as a model of chronic cerebral hypoperfusion; S212, after it is proved that the carotid stenosis patients can be used as chronic cerebral hypoperfusion models according to step S211, the MCI prevalence rates of the moderate carotid stenosis group, the severe carotid stenosis group and the control group are compared to prove whether the carotid stenosis is related to the prevalence of MCI, and further prove whether MCI can be used as an indicator for determining the chronic cerebral hypoperfusion model; S213. Comparison of the prevalence of WML in the moderate carotid stenosis group, severe carotid stenosis group and control group was used to prove whether the chronic cerebral hypoperfusion model is related to the prevalence of WML, and further prove whether WML can be used as an indicator to judge the chronic cerebral hypoperfusion model.

3. The method for analyzing the correlation between ANXA2 and chronic cerebral hypoperfusion blood-brain barrier destruction index and inflammatory injury index according to claim 2, characterized in that: Step S220 performs correlation analysis on MCI, blood-brain barrier and neuroinflammation during chronic cerebral hypoperfusion, including: S221. Correlation analysis was performed on the peripheral blood ANXA2 level, Claudin-5 level and IL-1β level in the MCI group and the non-MCI group to prove whether MCI is correlated with ANXA2 level, Claudin-5 level and IL-1β level, and then to determine whether the chronic cerebral hypoperfusion model is correlated with ANXA2 level, Claudin-5 level and IL-1β level; S222. When MCI is correlated with ANXA2 levels, Claudin-5 levels, and IL-1β levels, a multivariate logistic regression equation was constructed for the ANXA2 and IL-1β levels in peripheral blood serum. Multivariate binary logistic regression analysis was performed to determine whether ANXA2 and IL-1β were risk factors for MCI. Based on the analysis results, it was determined whether ANXA2 and IL-1β levels could be used to diagnose MCI.

4. The method for analyzing the correlation between ANXA2 and chronic cerebral hypoperfusion blood-brain barrier destruction index and inflammatory injury index according to claim 3, characterized in that: Step S230 performs correlation analysis on WML, blood-brain barrier and neuroinflammation during chronic cerebral hypoperfusion, including: S231. Correlation analysis was performed on the peripheral blood ANXA2 level, Claudin-5 level and IL-1β level in the mild WML group, moderate WML group and severe WML group to prove whether WML is correlated with ANXA2 level, Claudin-5 level and IL-1β level, and then to determine whether the chronic cerebral hypoperfusion model is correlated with ANXA2 level, Claudin-5 level and IL-1β level; S232. When WML is associated with ANXA2, Claudin-5 and IL-1β levels, a multivariate logistic regression equation was constructed for ANXA2, IL-1β and Claudin-5 levels in peripheral blood serum. Multivariate binary logistic regression analysis was used to demonstrate whether ANXA2 and IL-1β were risk factors for WML. Based on the analysis results, it was demonstrated whether ANXA2, IL-1β and Claudin-5 levels could be used to diagnose WML.

5. The method for analyzing the correlation between ANXA2 and chronic cerebral hypoperfusion blood-brain barrier destruction index and inflammatory injury index according to claim 4, characterized in that: In step S211, the correlation between the serum ANXA2, IL-1β and Claudin-5 levels in the carotid artery stenosis group and the control group was analyzed by rank sum test.

6. The method for analyzing the correlation between ANXA2 and chronic cerebral hypoperfusion blood-brain barrier destruction index and inflammatory injury index according to claim 4, characterized in that: In step S212, the chi-square test is used to analyze whether the chronic cerebral hypoperfusion model is narrow and the correlation between the degree of narrowing and the prevalence of MCI.

7. The method for analyzing the correlation between ANXA2 and chronic cerebral hypoperfusion blood-brain barrier destruction index and inflammatory injury index according to claim 4, characterized in that: In step S221 , the correlation between MCI and ANXA2 level, IL-1β level and Claudin-5 level was analyzed by Wilcoxon Z rank sum test.

8. The method for analyzing the correlation between ANXA2 and chronic cerebral hypoperfusion blood-brain barrier destruction index and inflammatory injury index according to claim 4, characterized in that: In step S231, the correlation between the severity of WML and the levels of ANXA2, Claudin-5 and IL-1β was analyzed by Kruskal-Wallis H rank sum test.

9. The method for analyzing the correlation between ANXA2 and chronic cerebral hypoperfusion blood-brain barrier destruction index and inflammatory injury index according to claim 4, characterized in that: In step S100, the carotid artery stenosis degree of the moderate stenosis group is 50%-69%, and the carotid artery stenosis degree of the severe stenosis group is 70%-99%.

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