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Application of Metrnl protein or gene in treatment of endothelial function impairment

A technology of endothelial function and protein, applied in the field of molecular biology and biomedicine, which can solve the problems of no report, Metrnl protein and few genes

Active Publication Date: 2017-08-25
SECOND MILITARY MEDICAL UNIV OF THE PEOPLES LIBERATION ARMY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] At present, there are very few reports on the function of Metrnl protein and gene, especially the relationship with endothelial function.

Method used

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  • Application of Metrnl protein or gene in treatment of endothelial function impairment
  • Application of Metrnl protein or gene in treatment of endothelial function impairment
  • Application of Metrnl protein or gene in treatment of endothelial function impairment

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0052] Example 1 Study on the expression and distribution of Metrn1 in wild mouse vascular endothelial cells

[0053] 1 Experimental materials

[0054] C57 mice: purchased from Shanghai Southern Model Biotechnology Development Co., Ltd.

[0055] Primary antibody Metrnl: purchased from Abcam.

[0056] Primary antibody CD31: purchased from Wuhan Google Biotechnology Co., Ltd.

[0057] Secondary anti-goat anti-rabbit, secondary anti-donkey anti-goat: purchased from Jackson Company.

[0058] Goat serum: purchased from Wuhan Boster.

[0059] DAPI: purchased from Biyuntian Company.

[0060] Anti-fluorescence quenching agent: purchased from Biyuntian Company.

[0061] High-sugar DMEM: purchased from Hyclone Company.

[0062] Type Ⅰ collagenase: purchased from Invitrogen Company.

[0063] EGM-2 special medium for endothelial cells: purchased from Lonza Company.

[0064] Trypsin: purchased from Gibco.

[0065] CD31 antibody with FITC fluorescent labeling: purchased from ebiosc...

Embodiment 2

[0093] Example 2 Construction and Identification of Metrnl Endothelial-specific Knockout Mice

[0094] 1 Experimental materials

[0095] Tek-cre mice: purchased from The Jackson Laboratory, B6.Cg-Tg(Tek-cre)12Flv / J (StockNumber: 004128).

[0096] Rat tail genome extraction kit: purchased from Zymo Company.

[0097] 2 Experimental methods

[0098] 2.1 Breeding of Metrnl endothelial-specific knockout mice

[0099] 2.1.1 Metrnl loxP / loxP mouse breeding

[0100] Metrnl loxP / loxP The mouse was constructed in the early stage of the laboratory and has been used to breed a variety of tissue-specific knockout mice. For specific breeding methods, please refer to the reference: Diabetes.2015Dec; 64(12):4011-22, and can also be obtained from the Purchased from the Second Military Medical University.

[0101] 2.1.2 Metrnl loxP / loxP Tek-cre cultivation

[0102] Will Metrnl loxP / loxP Mice were crossed with Tek-cre mice to obtain Metrnl loxP / wt Tek-cre mice and Metrnl loxP / loxP ...

Embodiment 3

[0119] Example 3 Detection of vasodilation function in Metrnl endothelial-specific knockout mice

[0120] 1 Experimental materials

[0121] Potassium chloride: purchased from Sinopharm Group.

[0122] Acetylcholine: purchased from Sigma.

[0123] Phenylephrine: purchased from Sigma.

[0124] Sodium nitroprusside: purchased from Sigma.

[0125] 2 Experimental methods

[0126] Metrnl endothelial-specific knockout mice (EC-Metrnl - / - ) and littermate control mice (WT) were subjected to thoracotomy under pentobarbital sodium anesthesia, the heart was cut open and bled, and the thoracic aorta was quickly separated and the thoracic aorta section was cut. There are 2 rings, which are carefully connected to the tension transducer, and the changes in vascular tension are continuously recorded through the recording system. Adjust the resting tension to a predetermined value of 1.5g; stabilize for 1 hour, keep the resting tension at 1.5g, and replace the K-H solution (37°C, contain...

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Abstract

The invention relates to application of Metrnl protein or gene in treatment of endothelial function impairment. On one hand, based on a Metrnl endothelial specific knockout mouse model, it is proved that due to lacking of Metrnl in blood vessel endothelia, an endothelium-dependent relaxation function is lowered, while Metrnl has the function of dilating blood vessels; on the other hand, based on a human umbilical vein endothelial cell line and Metrnl endothelial specific knockout mouse primary endothelial cells, it is proved that the cell proliferation is lowered after knockout of blood vessel endothelia Metrnl, showing that Metrnl has the effect of promoting endothelial cell proliferation. Therefore, the Metrnl protein or gene and a synergist of the Metrnl protein or gene can be used for preparing reagents for treating endothelium-dependent blood vessel relaxation dysfunction or regulating endothelial cell proliferation, and are applied to development of drugs of diseases related to endothelial function impariment.

Description

technical field [0001] The present invention relates to the fields of molecular biology and biomedicine, in particular to the application of Metrn1 protein or gene in the treatment of impaired endothelial function. Background technique [0002] Endothelial cells (ECs for short) are a thin layer of specialized epithelial cells, which are composed of a layer of flat cells in a polygonal shape, with jagged edges and interlocking cells. It is located between the blood and the vascular tissue, forms the inner wall of the blood vessel, and is the interface between the blood in the lumen of the blood vessel and the wall of the blood vessel. Endothelial cells line the entire circulatory system, from the heart down to the smallest capillaries. [0003] Endothelial cells provide a barrier between blood and vessel walls, and can complete the metabolic exchange of plasma and tissue fluid, and can also synthesize and secrete a variety of biologically active substances to ensure the norm...

Claims

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Application Information

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IPC IPC(8): A61K38/18A61K48/00A61P9/12A61P9/10A61P3/10A61P3/06A61P7/02A61P3/04
CPCA61K38/1866A61K48/00A61K2300/00
Inventor 缪朝玉宋捷李志勇管云枫
Owner SECOND MILITARY MEDICAL UNIV OF THE PEOPLES LIBERATION ARMY
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