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Application of minimal heterodimer partner in blood lipid regulation

A heterodimer and partner technology, applied in the field of treatment of hyperlipidemia, can solve problems such as inability to bind, not yet found, and lack of SHP

Inactive Publication Date: 2012-05-16
THE INST OF BASIC MEDICAL SCI OF CHINESE ACAD OF MEDICAL SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although highly similar to other nuclear receptor sequences, SHP is a relatively special member of the nuclear receptor family, and its ligand has not been found so far, so SHP is called an orphan receptor
In addition, SHP lacks the classical DNA-binding domain and cannot directly bind to DNA

Method used

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  • Application of minimal heterodimer partner in blood lipid regulation
  • Application of minimal heterodimer partner in blood lipid regulation
  • Application of minimal heterodimer partner in blood lipid regulation

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preparation example Construction

[0057] Preparation of Adeasier-1 Competent: Transform the Adeasy-1 plasmid into BJ5183, coat the plate (the culture plate contains Amp and streptomycin), pick a single clone to expand culture (the culture medium contains Amp and streptomycin), use 10% The BJ5183 electroporation competent was prepared by conventional method with glycerol, and stored at -80°C for later use. The table was digested with PmeI for 4 hours or overnight, fully linearized, and the digested DNA was purified with a gel recovery kit. The recovered linearized cells were transformed into Adeasier-1 BJ5183 electroporation-competent cells, spread on kanamycin-resistant LB plates, and cultured in a 37°C incubator for 14-16 hours. Pick 15-20 smaller clones, inoculate them in 5 ml of Kanna-resistant liquid LB medium, and shake overnight at 37°C. Harvest the bacteria in the next morning, extract the plasmid immediately (avoid storing the bacterial liquid at any temperature, but it can be made into glycerol bacte...

Embodiment 1

[0073]Experiments have shown that overexpression of HNF4 increases the promoter activity of ACAT2. In HuH7 cells, pGL3-hACAT2-1299, phRL-TK reporter gene plasmids were co-transfected with HNF4 expression plasmids or control plasmids. After 36 hours, the cells were collected and the activity of dual luciferase was detected. HNF4 overexpression significantly up-regulated the promoter activity of ACAT2 (see figure 1 ).

Embodiment 2

[0075] Further experiments demonstrated that adenoviral overexpression of HNF4 increased ACAT2 transcript levels. Adenovirus infection of HuH7 cells. Such as figure 2 As shown in Part A, the Western blot results showed that the HNF4 protein level in the adenovirus overexpression group (Ad-HNF4) was significantly higher than that in the control group (Ad-GFP). figure 2 Part B in the middle shows the real-time PCR results. The expression level of ACAT2 in the adenovirus overexpression group (Ad-HNF4) was about 5 times higher than that in the control group (Ad-GFP) when the adenovirus overexpressed HNF4.

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Abstract

The invention relates to an application of a minimal heterodimer partner in blood lipid regulation. Through the modes of reporter genes and adenovirus over expression, the inventor finds that a minimal heterodimer partner (SHP) can down-regulate the expression of acyl-CoA cholesterol acyltransferase 2 (ACAT 2), and thus finds that the minimal heterodimer partner polynucleotide molecule or protein is applicable to the preparation of medicaments for treating hyperlipidemia diseases. Through the reduction of ACAT 2 by SHP adenovirus over expression, a new approach for blood lipid regulation is provided.

Description

technical field [0001] The invention relates to the field of treatment of hyperlipidemia. Specifically, the present invention relates to the treatment of hyperlipidemia diseases by regulating fatty acyl-CoA cholesterol acyltransferase, an important regulator in lipid metabolism. Background technique [0002] The full name of ACAT is acyl-coA cholesterol acyltransferase (acyl-coA cholesterol acyltransferase), which catalyzes the esterification reaction, that is, adding fatty acyl groups to the hydroxyl groups of cholesterol to generate cholesterol esters. Cholesterol is a polar lipid molecule, while cholesteryl ester is a highly non-polar molecule. Therefore, the reaction catalyzed by ACAT greatly increases the hydrophobicity of cholesterol, and cholesteryl esters with strong hydrophobicity can accumulate with each other by means of hydrophobic interactions, which greatly increases the packing density of cholesteryl esters. [0003] The reaction catalyzed by ACAT has its im...

Claims

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Application Information

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IPC IPC(8): A61K48/00A61K38/17A61K31/7088A61P3/06A61P1/16A61P3/10A61P9/10A61P9/12
Inventor 刘德培张祝琴习杨杨瑞锋陈厚早
Owner THE INST OF BASIC MEDICAL SCI OF CHINESE ACAD OF MEDICAL SCI
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