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Method for detecting acetylated PINK1 in vascular endothelial cell

A technology of vascular endothelium and endothelial cells, applied in measuring devices, biological tests, material inspection products, etc., can solve the problems of inability to accurately detect vascular endothelial damage early and accurately reflect the functional status of endothelial cells

Inactive Publication Date: 2019-01-11
NINGBO UNIV
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AI Technical Summary

Problems solved by technology

Previously, the number of circulating endothelial cells (CEC) was used to measure the degree of damage to the vascular endothelium. The disadvantage of this method is that it cannot accurately reflect the functional status of endothelial cells.
Neither method can accurately detect early endothelial injury

Method used

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  • Method for detecting acetylated PINK1 in vascular endothelial cell
  • Method for detecting acetylated PINK1 in vascular endothelial cell
  • Method for detecting acetylated PINK1 in vascular endothelial cell

Examples

Experimental program
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Effect test

Embodiment Construction

[0017] (1) Acetylated PINK1 impairs mitophagy and leads to damage of vascular endothelial function

[0018] 1) Acetylated PINK1 interferes with PINK1 phosphorylation and impairs mitophagy

[0019] The mesenteric arterial tissue adjacent to the tumor resection was collected from colon cancer patients with diabetes mellitus and hyperlipidemia, the blood samples of the patients were collected, and the medical history data were collected. Establish high-fat feeding plus STZ-treated diabetic model mice and db / db diabetic model mice, and use immunohistochemistry, immunofluorescence, real-time PCR and Western Blot to detect patients with metabolic diseases such as diabetes and hyperlipidemia and type 2 diabetes model mouse vascular endothelial cell PCAF level and mtDNA copy number; observe the expression of PCAF induced by inflammatory factors such as IL-6 and ox-LDL and lipotoxic substances in HAEC cells, and observe the expression of PI3K / The activity status of AKT signaling path...

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Abstract

The invention discloses a semi-quantitative test paper for detecting an acetylated PINK1 protein in a vascular endothelial cell. The semi-quantitative test paper is mainly composed of five parts including a sample pad, a conjugate pad, a water absorption pad, a bottom plate and nitrocellulose membrane; two parallel antibody capturing lines are respectively drawn on the nitrocellulose membrane, theantibody capturing line close to the conjugate pad contains a concentration-determined acetylated PINK1 protein I monoclonal antibody in a first anti-vascular endothelial cell and is used as a detection line, the other antibody capturing line contains a concentration-determined goat anti-rabbit IgG antibody and is used as a standard concentration color control line, and the distance between the two lines is 4-7 mm; and the conjugate pad is sprayed with two substances including a colloidal gold or colored latex labeled acetylated PINK1 protein I monoclonal antibody in a second anti-vascular endothelial cell and a gold-labeled rabbit IgG.

Description

technical field [0001] The invention relates to a method for detecting acetylated PINK1 in vascular endothelial cells, in particular to a rapid and convenient fluorescent immunochromatographic rapid detection method and a semi-quantitative detection test paper for acetylated PINKI protein. Background technique [0002] The incidence of metabolic diseases in my country has risen rapidly this year, and these diseases are often the main culprits of vascular diseases, such as hyperlipidemia, diabetes and hyperuricemia. Studies have shown that the risk of cardiovascular disease in patients with type 2 diabetes (T2DM) is 2-4 times higher than that in non-diabetic patients, and the incidence of cardiovascular disease in middle-aged diabetics can be as high as 44%, and the death rate of cardiovascular diseases in diabetics The rate reached 2.7%. Adipose tissue has been re-recognized as a new endocrine organ. Obesity, especially central obesity, accumulates visceral fat that will se...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/68G01N33/531
CPCG01N33/6893G01N33/531
Inventor 步世忠陈晓敏方亮
Owner NINGBO UNIV
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