A biomarker for assessing the risk of lipid metabolism disorders and its application

Abstract

The present invention discloses a biomarker for assessing the risk of abnormal lipid metabolism and its application, which belongs to the field of molecular biology. The application of serum Sparcl1 as a biomarker for abnormal lipid metabolism in the preparation of a reagent for assessing the risk of abnormal lipid metabolism, and the method for assessing the risk of abnormal lipid metabolism includes: detecting the serum Sparcl1 expression level of the subject by a detection reagent or a kit, calculating the odds ratio (OR) and 95% confidence interval (CI) of abnormal lipid metabolism by multivariate logistic regression analysis, assessing the association between serum Sparcl1 protein and the risk of abnormal lipid metabolism, wherein a high serum Sparcl1 level indicates a low risk of abnormal lipid metabolism, and the serum Sparcl1 level is an independent risk factor for abnormal lipid metabolism, which can be used as a biomarker for abnormal lipid metabolism to assess the risk of abnormal lipid metabolism, and has important scientific and clinical value.

Description

Technical Field

[0001] The present invention belongs to the field of molecular biology, and in particular relates to a biomarker for assessing the risk of disease caused by abnormal lipid metabolism and its application. Background Art

[0002] Abnormal lipid metabolism is an important risk factor for type 2 diabetes and cardiovascular disease. In the past few decades, the prevalence of abnormal lipid metabolism in China has increased rapidly and has now reached 34.0%. More than half of Chinese adults have reduced high-density lipoprotein cholesterol (HDL-c), and 20.9% have elevated triglycerides (TG). The causes of abnormal lipid metabolism are multifaceted, including lifestyle and obesity. In-depth exploration of its mechanism of occurrence and the search for potential biomarkers have important scientific and clinical significance, and may provide effective therapeutic targets for disease intervention.

[0003] Sparcl1 is a secretory matrix cell glycoprotein. The gene is located on human chromosome 4q22. The gene size is 35kb, consisting of 11 exons and 10 introns. The mRNA length is 3kb, encoding 644 amino acids. The protein molecular weight is about 71kDa. Its molecular structure consists of three domains with specific functions, including the N-terminal acidic domain, the follistatin-like domain and the extracellular calcium ion binding domain, and there are multiple glycosylation sites. Sparcl1 is widely expressed in normal human tissues and can participate in a variety of physiological functions. It promotes lymphocyte transport by regulating the adhesion of endothelial cells and plays an important role in human growth and development. In recent years, studies have found that the Sparcl1 gene is closely related to tumorigenesis and is considered to be a potential tumor suppressor gene that participates in the occurrence and development of a variety of malignant tumors. It is known that Sparcl1 has different effects in different tumors. It is downregulated in non-small cell lung cancer, prostate cancer, ovarian cancer, colorectal cancer, gastric cancer, and pancreatic cancer, but upregulated in liver cancer and is closely related to tumor metastasis, recurrence and other prognoses. In addition, Sparcl1 may also be involved in the post-injury repair of Alzheimer's disease and ischemic stroke. The serum Sparcl1 level in the human population is independently associated with the severity of ischemic stroke. Animal experiments have found that after transient cerebral ischemia in adult rats, the expression of Sparcl1 in astrocytes is upregulated, which may be involved in angiogenesis and the occurrence and repair of synapses. However, there are currently few studies on whether Sparcl1 is involved in the physiological and pathological processes of metabolic diseases. Recent studies have reported that plasma Sparcl1 levels are elevated in patients with non-alcoholic steatohepatitis (NASH). Further studies have found that Sparcl1 activates the NF-κB / p65 signaling pathway by binding to toll-like receptor 4 (TLR4), increasing the expression of CC motif chemokine ligand 2 (CCL2), and thus participating in the progression of NASH. However, no studies have yet explored the role of Sparcl1 in the occurrence and development of abnormal lipid metabolism. Summary of the invention

[0004] The present invention provides application of Sparcl1 as a biomarker for abnormal lipid metabolism.

[0005] Preferably, Sparcl1 is used as a biomarker for abnormal lipid metabolism in the preparation of a reagent for assessing the risk of abnormal lipid metabolism.

[0006] The present invention also provides the use of a Sparcl1 detection reagent in preparing a reagent for assessing the risk of developing abnormal lipid metabolism. The Sparcl1 detection reagent is a reagent for detecting the expression level of Sparcl1 protein in a biological sample.

[0007] Preferably, the biological sample is serum, plasma or blood.

[0008] Preferably, the Sparcl1 detection reagent includes a reagent for detecting the expression level of Sparcl1 protein by immunohistochemistry, Western-Blot, protein immunoblotting, enzyme-linked immunosorbent assay (ELISA), flow cytometry, ultraviolet spectrophotometry-near infrared spectroscopy, high performance liquid chromatography, colorimetry or mass spectrometry.

[0009] The present invention also provides a detection reagent, comprising the Sparcl1 detection reagent.

[0010] The present invention also provides a detection kit, comprising the detection reagent.

[0011] The present invention also provides a method for assessing the risk of dyslipidemia, comprising the following steps:

[0012] (1) detecting the expression level of serum Sparcl1 protein in the subject using the detection reagent or the detection kit;

[0013] (2) Based on the Sparcl1 protein expression level detected in step (1), multivariate logistic regression analysis was used to calculate the odds ratio (OR) and 95% confidence interval (CI) of abnormal lipid metabolism, and to evaluate the association between serum Sparcl1 protein and the risk of abnormal lipid metabolism. A high serum Sparcl1 protein expression level indicates a low risk of abnormal lipid metabolism.

[0014] Preferably, the expression level of serum Sparcl1 protein is linearly negatively correlated with the risk of dyslipidemia, where the linear P = 0.0083, and the correlation trend between serum Sparcl1 protein and the reduced risk of dyslipidemia is consistent in each subgroup of the population (interaction P values ​​are all > 0.05), that is, the correlation between serum Sparl1 protein and the reduced risk of dyslipidemia does not change significantly due to the age, gender, smoking status, physical activity and body mass index level of the population.

[0015] Preferably, the OR value (95% CI) for the risk of dyslipidemia is 0.65 (0.49-0.87), and each standard deviation increase in serum Sparcl1 protein (501.40 ng / mL) is associated with a 14% decrease in the risk of dyslipidemia (OR, 0.86; 95% CI, 0.78-0.96).

[0016] The present invention has found through research that compared with normal people, the serum Sparcl1 level of patients with abnormal lipid metabolism is significantly reduced, and the prevalence of abnormal lipid metabolism is linearly negatively correlated with the serum Sparcl1 level, wherein the linear P=0.0083. The results of multivariate logistic regression show that after correction of confounding factors, the serum Sparcl1 level is negatively correlated with the risk of abnormal lipid metabolism, that is, a higher serum Sparcl1 level is significantly correlated with a lower risk of abnormal lipid metabolism. The serum Sparcl1 level is an independent risk factor for abnormal lipid metabolism and can be used as a biomarker for abnormal lipid metabolism to assess the risk of abnormal lipid metabolism, which has important scientific and clinical value. BRIEF DESCRIPTION OF THE DRAWINGS

[0017] Figure 1 This is the difference in expression of serum Sparcl1 levels between patients with abnormal lipid metabolism and a normal control group in the example.

[0018] Figure 2 It is the result of the prevalence of abnormal lipid metabolism grouped according to the serum Sparcl1 quartiles in the embodiment.

[0019] Figure 3 It is the restricted cubic spline between serum Sparcl1 and the risk of dyslipidemia in the embodiment; wherein, the solid line a represents the fitting relationship, the dotted line b represents the 95% confidence interval, the dotted line c represents the reference line of no association (odds ratio = 1.0), and area D represents the density distribution of serum Sparcl1. The model corrects indicators such as age, gender, smoking status, drinking status, physical activity, education level and body mass index (BMI).

[0020] Figure 4 This is a subgroup analysis of the association between each SD increment of serum Sparcl1 and the risk of dyslipidemia in the embodiment, and the model corrects indicators such as age, gender, smoking status, drinking status, physical activity, education level and BMI (except stratification factors). DETAILED DESCRIPTION

[0021] In order to make the purpose, technical solution and advantages of the embodiment of the present invention clearer, the technical solution of the embodiment of the present invention will be clearly and completely described in conjunction with the drawings of the embodiment of the present invention. Obviously, the described embodiment is a part of the embodiment of the present invention, not all of the embodiments. Based on the described embodiment of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work belong to the scope of protection of the present invention.

[0022] Example 1

[0023] This example includes 1633 people from the Shanghai community, and serum Sparcl1 levels were measured by enzyme-linked immunosorbent assay. Abnormal lipid metabolism was defined using the National Cholesterol Education Program Adult Treatment Panel III criteria, and patients were defined as having abnormal lipid metabolism if they had any of the following: (a) total cholesterol ≥6.22mmol / L; (b) low-density lipoprotein cholesterol ≥4.14mmol / L; (c) triglycerides ≥2.26mmol / L; (d) high-density lipoprotein cholesterol <1.04mmol / L. Multivariate logistic regression analysis was used to calculate the odds ratio (OR) and 95% confidence interval (CI) between the fourth and lowest quartiles of abnormal lipid metabolism, thereby evaluating the association between serum Sparcl1 levels and the risk of abnormal lipid metabolism. The model adjusted for age, gender, smoking status, drinking status, physical activity, education level, and body mass index (BMI). SAS 9.4 was used for statistical analysis, and a two-sided P value of <0.05 was statistically significant. The results are as follows Figure 1-4 , as shown in Table 1 and Table 2.

[0024] Among the 1633 participants included, the mean (SD) age was 55.66±8.34, of whom 577 (35.33%) had dyslipidemia, and the average serum Sparcl1 level was 701.92±501.40 ng / mL. The comparison of serum Sparcl1 levels between patients with dyslipidemia and the normal control group showed that the serum Sparcl1 level in patients with dyslipidemia was significantly lower than that in normal controls ( Figure 1 ).

[0025] The baseline characteristics of the population according to the serum Sparcl1 quartiles are shown in Table 1: As the serum Sparcl1 level increased, the triglyceride level of the participants gradually decreased, while the high-density lipoprotein cholesterol (HDL-C) level increased (trends P < 0.05; Table 1). The results of the prevalence of abnormal lipid metabolism grouped by serum Sparcl1 quartiles showed that as the serum Sparcl1 level increased, the prevalence of abnormal lipid metabolism decreased significantly. The prevalence of abnormal lipid metabolism from the 1st quartile to the 4th quartile was 40.69%, 35.05%, 34.72%, 30.88% ( Figure 2 ).

[0026] Table 1: Basic information of the subjects grouped according to serum Sparcl1 quartile levels

[0027]

[0028] The results of logistic regression analysis showed that compared with the participants in the lowest quartile of serum Sparcl1, the risk of dyslipidemia in the highest quartile of serum Sparcl1 was significantly reduced, with an OR value (95% CI) of 0.65 (0.49-0.87) for the risk of dyslipidemia. Each standard deviation increase in serum Sparcl1 (501.40 ng / mL) was associated with a 14% decrease in the risk of dyslipidemia (OR, 0.86; 95% CI, 0.78-0.96). After further adjustment for age, gender, lifestyle factors and BMI, the correlation between serum Sparcl1 levels and the risk of lipid metabolism was still significant. The results of each component of abnormal lipid metabolism showed that the participants in the highest quartile of serum Sparcl1 had significantly reduced risks of elevated triglycerides (OR, 0.54; 95% CI, 0.35-0.83) and reduced high-density lipoprotein cholesterol (OR, 0.67; 95% CI, 0.45-0.99), but had no significant association with the risk of elevated total cholesterol and low-density lipoprotein cholesterol (Table 2). After adjusting for age, sex, smoking status, drinking status, physical activity, education level, and body mass index, the restricted cubic spline results between serum Sparcl1 and the risk of abnormal lipid metabolism showed that serum Sparcl1 was linearly negatively correlated with the risk of abnormal lipid metabolism, with a linear P = 0.0083 ( Figure 3 ); The results of stratified analysis showed that the correlation trend between serum Sparl1 and the reduced risk of dyslipidemia was consistent in all subgroups (interaction P values ​​were all > 0.05), that is, the relationship between serum Sparl1 and the risk of dyslipidemia did not change significantly due to age, gender, smoking status, physical activity and body mass index level ( Figure 4 ).

[0029] Table 2: Correlation between serum Sparcl1 levels and the risk of dyslipidemia

[0030]

[0031] In summary, compared with normal people, the serum Sparcl1 level of patients with abnormal lipid metabolism was significantly lower, and the prevalence of abnormal lipid metabolism was negatively correlated with the serum Sparcl1 level; after adjusting for confounding factors, the serum Sparcl1 level was negatively correlated with the risk of abnormal lipid metabolism, and a higher serum Sparcl1 level was significantly correlated with a reduced risk of abnormal lipid metabolism, indicating that serum Sparcl1 level is an independent risk factor for abnormal lipid metabolism, and can be used as a biomarker for abnormal lipid metabolism to assess the risk of abnormal lipid metabolism, or can provide a potential effective therapeutic target for disease intervention, which has important scientific and clinical value.

[0032] The above description of the embodiments is to facilitate the understanding and use of the present invention by those skilled in the art. It is obvious that those skilled in the art can easily make various modifications to these embodiments and apply the general principles described herein to other embodiments without creative work. Therefore, the present invention is not limited to the above embodiments. Improvements and modifications made by those skilled in the art based on the principles of the present invention without departing from the scope of the present invention should be within the protection scope of the present invention.

Claims

1. Application of Sparcl1 as a biomarker of abnormal lipid metabolism in the preparation of reagents for assessing the risk of abnormal lipid metabolism.

2. Use of a Sparcl1 detection reagent in the preparation of a reagent for assessing the risk of developing abnormal lipid metabolism, wherein the Sparcl1 detection reagent is a reagent for detecting the expression level of Sparcl1 protein in a biological sample.

3. The use according to claim 2, characterized in that: The biological sample is serum, plasma or blood.

4. The use according to claim 2, characterized in that: The Sparcl1 detection reagent includes a reagent for detecting the expression level of Sparcl1 protein by immunohistochemistry, Western-Blot, protein immunoblotting, enzyme-linked immunosorbent assay, flow cytometry, ultraviolet spectrophotometry-near infrared spectroscopy, high performance liquid chromatography, colorimetry or mass spectrometry identification.

5. The use according to claim 2, characterized in that: The method for evaluating the risk of abnormal lipid metabolism by using the Sparcl1 detection reagent comprises the following steps: (1) using the serum of the subject as a biological sample, and detecting the expression level of the serum Sparcl1 protein of the subject using the Sparcl1 detection reagent; (2) Based on the serum Sparcl1 protein expression level detected in step (1), multivariate logistic regression analysis was used to calculate the OR and 95% CI of abnormal lipid metabolism, and the association between the serum Sparcl1 protein expression level and the risk of abnormal lipid metabolism was evaluated, where a high serum Sparcl1 protein expression level indicated a low risk of abnormal lipid metabolism.

6. The use according to claim 2, characterized in that: The expression level of serum Sparcl1 protein was linearly negatively correlated with the risk of dyslipidemia, with linear P=0.0083. The correlation trend between serum Sparcl1 protein and the reduced risk of dyslipidemia was consistent in each subgroup, and the interaction P values ​​were all greater than 0.

05.

7. The use according to claim 2, characterized in that: The OR value of the risk of dyslipidemia in the fourth quartile and the lowest quartile of serum Sparcl1 protein level was 0.65, 95% CI: 0.49-0.

87. Every 501.40 ng / mL increase in serum Sparcl1 protein level was associated with a 14% decrease in the risk of dyslipidemia, OR: 0.86; 95% CI: 0.78-0.96.

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