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Biomarker for predicting II-type diabetic vascular complication risk and application thereof

A technology for diabetes and complications, applied in cardiovascular system diseases, biological tests, biochemical equipment and methods, etc., can solve problems such as inability to choose treatment options, high mortality, and poor treatment effects of cardiovascular and cerebrovascular complications

Inactive Publication Date: 2017-06-30
FUDAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

As mentioned above, cardiovascular complications are the main cause of death in diabetes, and antiplatelet drugs are the main means to prevent and treat cardiovascular complications of diabetes. However, the reason for the increased platelet reactivity in diabetic patients is not clear, so it is not yet possible in clinical practice. The selection of targeted treatment options and drugs, as well as the inability to carry out precise treatment, lead to poor treatment effect and high mortality rate of type II diabetes mellitus and cerebrovascular complications

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  • Biomarker for predicting II-type diabetic vascular complication risk and application thereof
  • Biomarker for predicting II-type diabetic vascular complication risk and application thereof
  • Biomarker for predicting II-type diabetic vascular complication risk and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0034] Example 1. Detection of P2Y12 receptor mRNA expression level in platelets of type II diabetic subjects by real-time fluorescent quantitative PCR method

[0035] The purpose of this example is to prove that the expression level of P2Y12 receptor mRNA in platelets of subjects with type II diabetes is significantly higher than that in platelets of healthy subjects. In this example, the preparation of platelets from patients with type II diabetes and healthy subjects: the platelets come from volunteers who have signed the informed consent form, and the volunteers have not taken any antiplatelet drugs such as aspirin or clopidogrel within 20 days before blood collection. drug. The anticoagulant was ACD (85mmol L-1 sodium citrate, 71.38 mmol L-1 citric acid, and 27.78mmol L-1 glucose). Centrifuge at 300g for 20 minutes to obtain platelet-rich plasma, and then centrifuge at 900g for 10 minutes to obtain platelets after taking the supernatant. The total mRNA was extracted by ...

Embodiment 2

[0038] Example 2. Detection of P2Y12 receptor expression level in platelets of type II diabetic subjects by Western Blot method

[0039] The purpose of this example is to prove that the expression level of P2Y12 receptor protein in platelets of subjects with type II diabetes is significantly higher than that in platelets of healthy subjects.

[0040] In this example, the preparation method of platelets from type II diabetic subjects and healthy subjects is the same as that in Example 1. The platelets were lysed and resuspended by adding SDS lysis buffer, boiled for later use. The expression level of P2Y12 receptor protein was determined by Western Blot western blotting method, and the formula was:

[0041] P2Y12 receptor protein expression level=P2Y12 receptor protein band density / GAPDH protein band density

[0042] The results of the expression level of P2Y12 receptor protein in the subject's platelet samples are as follows: figure 2 As shown, the P2Y12 receptor protein e...

Embodiment 3

[0043] Example 3. ADP-induced platelet aggregation rate of platelet samples from type II diabetic subjects

[0044] The purpose of this example is to prove that the platelet aggregation rate of subjects with type II diabetes is significantly higher than that of healthy subjects.

[0045] In this example, the preparation method of platelets from type II diabetic subjects and healthy subjects is the same as that in Example 1. Platelets were resuspended in Tyrode buffer (138mmol L -1 NaCl, 2.7mmolL -1 KCl, 2 mmol L -1 MgCl 2 ,0.42mmol L -1 NaH 2 PO 4 ,5mmolL -1 Glucose, 10mmol L -1 HEPES, 0.2% bovine serum albumin, and 0.02 unit mL -1 apyrase, pH7.4), count platelets, and adjust the number of platelets to 2.5×10 8 platelets / mL;

[0046] Determination of platelet aggregation: Platelet aggregation was performed in a platelet aggregation instrument (Model 400VS, Chrono-Log, Haverston, PA), parameter settings: stirring speed 900rpm, temperature 37°C, before adding different ...

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Abstract

The invention belongs to the technical field of biology and diagnosis, and relates to a receptor of a definite, effective and visualized platelet reactivity biomarker P2Y12 and an application thereof for diagnosing and predicting the II-type diabetic vascular complication risk. The platelet activation biomarker P2Y12 can detect expression levels of platelet through fluorescent quantitative polymerase chain reaction (PCR), Western blot, enzyme linked immunosorbent assay (ELISA), flow cytometry, UV spectrophotometry-near infrared spectrometry, high performance liquid chromatography, colorimetry, a mass spectrum identification method and the like and detect content of P2Y12 in whole blood. The invention also provides a method of biomarker P2Y12 for diagnosing and predicting vascular complication of subjects as well as a medicine for treating II-type diabetic vascular complication and a screening method of the medicine. The invention further provides an intervene plane for individualizing P2Y12 inverse agonist and accurately treating the vascular complication of II-type diabetic patients.

Description

technical field [0001] The invention belongs to the technical field of biology and diagnostics, and specifically relates to the technical field of prediction and treatment of type II diabetes cardiovascular complications risk. In particular, it relates to biomarkers for diagnosing and predicting the risk of cardiovascular complications in patients with type II diabetes; and methods for diagnosing and predicting the risk of cardiovascular complications in subjects. The present invention further relates to drugs for the treatment of cardiovascular complications in patients with type II diabetes (increased risk of suffering from this disease), and methods for screening such drugs. Background technique [0002] In recent years, with the improvement of living standards and population aging in my country, the incidence and mortality of type II diabetes have also increased year by year. According to statistics, the prevalence of type II diabetes among adults in my country is about...

Claims

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Application Information

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IPC IPC(8): C12Q1/68G01N33/68G01N15/14G01N21/33G01N21/359G01N30/02G01N27/62G01N21/00A61K31/7064A61P3/10A61P9/00
CPCA61K31/7064C12Q1/686G01N15/14G01N21/00G01N21/33G01N21/359G01N27/62G01N30/02G01N33/68C12Q2563/107C12Q2545/114C12Q2531/113
Inventor 丁忠仁胡亮张艳张思
Owner FUDAN UNIV
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