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Application of tgf-β1 expressed by dendritic cells in the preparation of anti-atherosclerotic drugs

A technology of atherosclerosis and dendritic cells, applied in the medical field, can solve problems that are unclear and affect the formation of atherosclerotic plaques

Inactive Publication Date: 2016-11-16
TAISHAN MEDICAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Studies have reported that DC also has the effect of lowering blood lipids. It is not clear whether DC lowers blood lipids through the expression of TGF-β1 and ultimately affects the formation of atherosclerotic plaques.

Method used

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  • Application of tgf-β1 expressed by dendritic cells in the preparation of anti-atherosclerotic drugs
  • Application of tgf-β1 expressed by dendritic cells in the preparation of anti-atherosclerotic drugs
  • Application of tgf-β1 expressed by dendritic cells in the preparation of anti-atherosclerotic drugs

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0018] Example 1 Using cre-lox technology to construct DCs that do not express TGF-β1 mice

[0019] The principle of cre-lox technology is: Cre is a recombinase that can catalyze the recombination of two loxp sites, thereby deleting the gene between the loxp sites. In order to use the cre-lox technology to construct mice whose DCs do not express TGF-β1, two kinds of mice, CD11c cre (Catalog No.: 007567) and TGF-β1 flox mice (product number: 010721), and then use the principle of cre-lox technology to cross the two kinds of mice. The principle of hybridization is as follows: figure 1 . The F1 generation mice were identified by PCR, and the genotypes of the offspring were selected as CD11c-cre (transgenic) and TGF-β1-flox (heterozygous) mice and then crossed; in the F2 generation mice, CD11c-cre was selected as the transgene The mice that are homozygous for TGF-β1-flox are then crossed to expand breeding; CD11c-cre (transgenic, hemizygous) and TGF-β1-flox (homozygous) in the ...

Embodiment 2

[0059] Example 2 Using bone marrow chimeric technology to construct Apoe in which DC does not express TGF-β1 - / - mouse

[0060] Apoe - / - Mice are susceptible mice for atherosclerosis. In this experiment, Apoe - / - The mouse is the recipient mouse, and the CD11c constructed above cre -TGF-β1 flox The mice were used as donor mice, and through bone marrow chimera technology, Apoe in which DCs did not express TGF-β1 was established. - / - mice. This technique first aseptically washes out the above-mentioned constructed CD11c cre -TGF-β1 flox Mouse bone marrow, preparation of bone marrow cells due to TGF-β1 flox Genetic background of mice and Apoe - / - Mice have different genetic backgrounds. In order to prevent transplant rejection during bone marrow transplantation, the prepared bone marrow cells are further removed by magnetic bead sorting technology to remove T cells. The bone marrow cells sorted by magnetic beads were reinfused into Apoe irradiated with 12Gray lethal dos...

Embodiment 3

[0083] Example 3 Induction of Apoe through bone marrow chimerism by high-fat diet - / - Atherosclerosis in mice

[0084]The normal feed was fed for 6 weeks after bone marrow chimerism, so that the transplanted bone marrow cells could be rebuilt. After 6 weeks, a high-fat diet (containing 0.15% cholesterol and 21% fat) was continued for 12 weeks to induce the formation of atherosclerotic plaques in mice. The feed was Co60 irradiated and purchased from Beijing Keao Xieli Feed Co., Ltd. At 12 weeks, the plasma total cholesterol and triglyceride levels, atherosclerotic plaque area and plaque / lumen ratio in the aortic root were detected. The specific methods and results are as follows.

[0085] 1. Reagent preparation

[0086] PBS (pH7.2, 0.01M) was prepared by conventional methods; EDTA anticoagulant was prepared by conventional methods; Oil Red O staining kit was purchased from Nanjing Jiancheng Bioengineering Institute; OCT used for embedding tissues was American Cherry Blossom...

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Abstract

The invention discloses an application of TGF-β1 expressed by dendritic cells in the preparation of anti-atherosclerosis drugs. In the present invention, firstly, through cre-lox technology, a mouse in which dendritic cells (DC) do not express TGF-β1 is successfully hybridized, and the bone marrow of this mouse is chimerized into an atherosclerosis-susceptible mouse by bone marrow transplantation Apoe ‑ / ‑ , induced atherosclerosis in mice by a high-fat diet. It was found that DCs not expressing TGF‑β1 caused an increase in total plasma cholesterol and a significant increase in the area of ​​atherosclerotic plaque. These results indicated that TGF‑β1 expressed by DCs could play an anti-atherosclerotic effect by reducing plasma cholesterol levels. Based on the anti-atherosclerosis effect of TGF-β1 expressed by DCs, the TGF-β1 expressed by DCs can be used to prepare anti-atherosclerotic drugs, which provides a new direction for the development of anti-atherosclerotic drugs.

Description

technical field [0001] The invention belongs to the field of medicine and relates to the application of TGF-β1 expressed by dendritic cells in the preparation of anti-atherosclerotic drugs. Background technique [0002] TGF-β can be secreted by a variety of cells, and its family includes three members, TGF-β1, 2 and 3. The most predominant isoform expressed by the immune system is TGF-β1. A large number of studies have shown that the expression of TGF-β1 is related to the formation of atherosclerosis. Some studies have shown that TGF-β1 is an anti-atherosclerotic vascular protective cytokine, which can limit the growth of atherosclerotic plaques, prevent aortic expansion and stabilize plaque structure. However, some studies also believe that TGF-β has the effect of promoting atherosclerosis, and found that TGF-β1 can accelerate atherosclerosis by increasing the accumulation of extracellular matrix and lipid retention in blood vessels. The above research results suggest th...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K38/18A61P9/10
Inventor 李雅林唐华宋文刚刘有旺
Owner TAISHAN MEDICAL UNIV
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