Marker for atherosclerosis diagnosis and application thereof

Abstract

The invention discloses a marker for atherosclerosis diagnosis and application thereof, and belongs to the field of biological medicine. The marker is GS-23alpha, and the marker is applied to preparation of a detection reagent for diagnosing atherosclerosis. The detection reagent is used for quantitatively detecting the expression quantity of the marker GS-23alpha. Experimental results show that the expression level of GS-23alpha is consistent with the severity of atherosclerosis, and it can be further proved that GS-23alpha can be used as a marker for detecting the atherosclerosis process.

Description

Technical Field

[0001] The present invention belongs to the field of biomedicine, and particularly relates to a biomarker for the diagnosis of atherosclerosis and its application. Background Art

[0002] Atherosclerosis (AS) is a chronic inflammatory disease, and its typical feature is the formation of fibro-fatty lesions under the arterial intima. AS is the most important cause of cardiovascular, cerebrovascular and peripheral vascular diseases (such as myocardial infarction, ischemic stroke, lower limb gangrene, etc.). The susceptibility of AS in different parts of the human arterial system is significantly different. Among them, the abdominal aorta, iliac artery, carotid artery and coronary artery are the four most susceptible parts of AS, while the subclavian artery, aortic arch and thoracic aorta are not prone to AS. Due to the lack of an ideal large animal model that can simulate human AS, the understanding of the characteristics and pathogenesis of AS-susceptible parts is currently very limited. Constructing an ideal large animal model similar to the human AS lesion site and comparing the characteristic differences, cell composition differences, cell proportion differences and gene expression differences between AS-susceptible parts and non-susceptible parts is an effective way to elucidate its pathogenesis. Further exploring biomarkers that can be used for the clinical diagnosis of atherosclerosis is of great significance. Summary of the Invention

[0003] The present invention provides a biomarker for the diagnosis of atherosclerosis and its application.

[0004] The present invention adopts the following technical solutions: On the one hand, the present invention relates to a biomarker for the diagnosis of atherosclerosis, and the biomarker is GS-23α, and its amino acid sequence is as shown in SEQ ID No.1, specifically Ala-Cys-Asp-His-Pro-Arg-Met-Glu-Cys-Asp-Ser-Ile-Asp-Met-Tyr-Asp-Asp-Asp-Ser-Lys-Asp-Ser-Asp-Ser-Lys-His-Asp-Ser-Leu-Lys-Asp-Ala-Tyr-Asp-Ser-Ala-Trp-Ala-Met-Pro-Glu-Arg-Cys-Thr-Tyr-Tyr-Asp-Tyr-Ala-Ser-Pro-Arg-Ser-Ala-Lys-Asn-Met-Asp-Pro-Glu-Pro-Ser-Ala-Ala-Ala-Pro-Glu-Met-Lys-Ser-Ile-Trp-Asp-Ser-Lys-Phe-Ala-Asp-Phe-Pro-Phe-Ala-Asp-Ser-Phe-Pro-Asp-Ala-Ser-Phe-Pro-Asp-Ser-Ala-Ile-Phe-Pro-Asp-Ser。

[0005] On the other hand, the present invention also relates to the use of the above-mentioned marker GS-23α in the preparation of a detection reagent for diagnosing atherosclerosis.

[0006] In a preferred embodiment of the present invention, the detection reagent is used for quantitatively detecting the expression level of the marker GS-23α.

[0007] In a preferred embodiment of the present invention, the detection reagent is used for quantitatively detecting the expression level of the marker GS-23α in peripheral blood cells.

[0008] In a preferred embodiment of the present invention, the detection reagent is a kit, a gene chip or a test strip.

[0009] On the other hand, the present invention also relates to a detection reagent which is used for quantitatively detecting the expression level of the marker GS-23α in peripheral blood cells.

[0010] Beneficial effects: Experimental results show that the expression level of GS-23α is consistent with the severity of atherosclerosis, which can further prove that it can be used as a marker for detecting the process of atherosclerosis. Description of the Drawings

[0011] Figure 1 It is a result diagram showing the high expression of GS-23α in atherosclerotic plaques in the examples of the present invention; Figure 2 It is a comparison diagram of the expression levels of GS-23α in peripheral blood monocytes of atherosclerotic patients and normal people. Detailed Embodiments

[0012] To facilitate the understanding of the present invention, a more comprehensive description will be given below, and preferred embodiments of the present invention will be provided. However, the present invention can be implemented in many different forms and is not limited to the embodiments described herein.

[0013] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by those skilled in the technical field to which the present invention belongs. Unless otherwise specified, various materials and reagents used in the examples are common materials and reagents in the art and can be obtained through conventional commercial channels; the methods used are all conventional methods well-known to those skilled in the art.

[0014] Example 1: High expression of GS-23α in atherosclerotic plaques Twenty C57BL / 6 mice on a normal diet, twenty C57BL / 6 mice on a high-fat diet, twenty spontaneous atherosclerotic ApoE - / - mice on a normal diet, and twenty spontaneous atherosclerotic ApoE - / - After one week of adaptation, the mice on a high-fat diet were fed with a high-fat diet (containing 0.25% cholesterol). After five weeks of feeding, the aortic tissues of the mice were isolated, and total RNA of the tissues was extracted. The mRNA level of GS-23α was detected by real-time quantitative PCR. The C57BL / 6 mice on a normal diet and the C57BL / 6 mice on a high-fat diet did not show obvious atherosclerotic plaques. Obvious atherosclerotic plaques had appeared in the aortas of the ApoE - / - mice on a normal diet, and the atherosclerotic degree of the aortas of the ApoE - / - mice on a high-fat diet was more severe.

[0015] The detection was carried out as described in Figure 1 , and the detection results showed that, compared with the tissues of C57BL / 6 mice on a normal diet without atherosclerotic plaques and C57BL / 6 mice on a high-fat diet, the mRNA level of GS-23α was significantly increased in the tissues of ApoE - / - mice on a normal diet with atherosclerotic plaques, and the expression level was further increased in the aortic tissues of ApoE - / - mice on a high-fat diet. P < 0.05, P < 0.01; n.s., no difference. The p value was calculated by the Student's t test for statistical analysis, and the data were expressed in the form of mean plus or minus standard error. The above results suggest that the expression level of GS-23α is consistent with the severity of atherosclerosis and is expected to be used as a biomarker for the detection of the atherosclerotic process.

[0016] Example 2: Detecting and comparing the expression levels of GS-23α in peripheral blood mononuclear cells of normal individuals and patients with atherosclerosis Peripheral blood mononuclear cells were isolated from the lymphocyte separation medium (purchased from Tianjin Haoyang Biological Company) of normal individuals (10 cases as the control group) and patients with atherosclerosis (10 cases as the experimental group). After centrifugation, the buffy coat in the middle was aspirated. Then, the cells were washed twice with 0.9% sodium chloride solution and resuspended in X-VIVO culture medium containing 10% PBS, and the cell concentration was adjusted to 1×10 6 cells / mL. The expression of GS-23α was detected by qRT-PCR, and the detection results are as Figure 2 shown.

[0017] As Figure 2 can be seen, there were significant differences in the expression of GS-23α in peripheral blood mononuclear cells between healthy individuals (NC) and patients with atherosclerosis (AS) (P<0.05), and the expression of GS-23α in peripheral blood mononuclear cells of patients with atherosclerosis showed an obvious upward trend. The above results suggest that the expression level of GS-23α is consistent with the severity of atherosclerosis, which can further prove that it can be used as a marker for detecting the process of atherosclerosis.

[0018] The preferred embodiments of the present invention have been described above, but they are not intended to limit the present invention. Those skilled in the art can make improvements and changes to the embodiments disclosed herein without departing from the scope and spirit of the present invention.

Claims

1. A marker for the diagnosis of atherosclerosis, characterized in that: The marker is GS-23α, and its amino acid sequence is specifically as follows: Ala-Cys-Asp-His-Pro-Arg-Met-Glu-Cys-Asp-Ser-Ile-Asp-Met-Tyr-Asp-Asp-Asp-Ser-Lys-Asp-Ser-Asp-Ser-Lys-His-Asp-Ser-Leu-Lys-Asp-Ala-Tyr-Asp-Ser-Ala-Trp-Ala-Met-Pro-Glu-Arg-Cys-Thr-Tyr-Tyr-Asp-Tyr-Ala-Ser-Pro-Arg-Ser-Ala-Lys-Asn-Met-Asp-Pro-Glu-Pro-Ser-Ala-Ala-Ala-Pro-Glu-Met-Lys-Ser-Ile-Trp-Asp-Ser-Lys-Phe-Ala-Asp-Phe-Pro-Phe-Ala-Asp-Ser-Phe-Pro-Asp-Ala-Ser-Phe-Pro-Asp-Ser-Ala-Ile-Phe-Pro-Asp-Ser.

2. Use of the marker according to claim 1 in the preparation of a detection reagent for diagnosing atherosclerosis.

3. The application according to claim 2, wherein: The detection reagent is used for quantitatively detecting the expression level of the marker GS-23α.

4. The application according to claim 2, wherein: The detection reagent is used for quantitatively detecting the expression level of the marker GS-23α in peripheral blood cells.

5. The application according to claim 2, characterized in that: The detection reagent is a kit, a gene chip or a test strip.

6. A detection reagent, characterized in that: The detection reagent is used for quantitatively detecting the expression level of the marker GS-23α in peripheral blood cells.

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