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TRAF4 (Tumor Necrosis Factor Receptor-associated Factor 4) and application of inhibitor thereof in preparing medicine for treating fatty liver and associated diseases

A technology of tumor necrosis factor and related factors, which is applied in the direction of drug combination, pharmaceutical formula, medical preparations containing active ingredients, etc.

Active Publication Date: 2018-07-27
WUHAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there is no report on the direct relationship between TRAF4 and fatty liver and related diseases

Method used

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  • TRAF4 (Tumor Necrosis Factor Receptor-associated Factor 4) and application of inhibitor thereof in preparing medicine for treating fatty liver and associated diseases
  • TRAF4 (Tumor Necrosis Factor Receptor-associated Factor 4) and application of inhibitor thereof in preparing medicine for treating fatty liver and associated diseases
  • TRAF4 (Tumor Necrosis Factor Receptor-associated Factor 4) and application of inhibitor thereof in preparing medicine for treating fatty liver and associated diseases

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0114] [Example 1] In the human liver cell line L02 cells, a stable transfection cell line of TRAF4 knockdown (shTRAF4) was established

[0115] According to the steps of establishing the L02 stable transgenic cell line in the embodiment, a stable transgenic cell line with TRAF4 knockdown (shTRAF4) was established. Afterwards, the cells were collected, and the expression of TRAF4 was verified by Western Blot. The result is as figure 1 As shown, it can be seen that in L02 cells infected with the shTRAF4 lentivirus system, the expression of TRAF4 was significantly reduced, indicating that the cell line was established successfully.

Embodiment 2

[0116] [Example 2] In the human liver cell line L02 cells, the effect of TRAF4 knockdown on hepatic fat accumulation

[0117] L02 cells were divided into 6 groups: pLKO.1 control group, shTRAF4 control group, pLKO.1 experimental group×2, shTRAF4 experimental group×2. After the cells adhered to the wall, palmitate (PA) and oleic acid (OA) (PA 0.1mM+OA 0.2mM), (PA 0.2mM+OA 0.4mM) were added to the experimental group for stimulation, and the same amount of BSA, oil red O staining after 12h.

[0118] The results of Oil Red O staining were as follows: figure 2 As shown, the cells in the control group had no obvious staining, but after adding PA+OA stimulation, the cells stained by Oil Red O increased significantly compared with the control group, and the increase in the staining area of ​​the shTRAF4 experimental group was smaller than that of the pLKO.1 experimental group . The results indicated that the reduction of TRAF4 expression could inhibit the lipid accumulation in PA-...

Embodiment 3

[0119] [Example 3] In primary mouse hepatocytes, the effect of TRAF4 knockdown on fat accumulation in primary hepatocytes 1) Three adenoviruses constructed by Hanheng Company were used to infect primary mouse hepatocytes, and Western Blot was completed Detect the expression of TRAF4, the result is as follows image 3 As shown, it can be seen that the adenovirus effect of shTRAF4-3 is the most obvious.

[0120] 2) The mouse primary hepatocytes were divided into two groups: NC group and shTRAF4-3 group. After 36 hours of adenovirus infection, palmitate (PA) and oleic acid (OA) (PA 0.1 mM+OA 0.2 mM) were added, and oil red O staining was performed after 12 hours of action.

[0121] The results of Oil Red O staining were as follows: Figure 4 As shown, after adding PA+OA stimulation, the cells in the NC group were significantly stained, while the stained area of ​​the shTRAF4-3 group was significantly smaller than that in the NC group. The results indicated that the reduction o...

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Abstract

The invention discloses a TRAF4 (Tumor Necrosis Factor Receptor-associated Factor 4) and application of an inhibitor thereof in preparing medicine for treating fatty liver and associated diseases. Bytaking a human normal liver L02 cell line, a TRAF2 knocked-down L02 cell line, primary hepatocyte and TRAF4 knocked-down primary hepatocyte as research objects and stimulating and inducing a liver cell lipid accumulation model by PA (Palmitic Acid) and OA (Oleic Acid), functions of a TRAF4 gene can be researched. The research discovers that when expression of the TRAF4 is reduced, L02 cell lipid accumulation stimulated and induced by the PA and the OA is remarkably reduced, i.e., the TRAF4 gene is capable of promoting occurrence and development of the fatty liver and the associated diseases, so that the TRAF4 provides a target for developing the medicine for preventing, relieving and / or treating the fatty liver and the associated diseases.

Description

technical field [0001] The invention belongs to the field of gene function and application, in particular to the function of a tumor necrosis factor receptor-associated factor 4 (Tumor necrosis factor receptor-associated factor 4, TRAF4) as a drug target in the screening of drugs for the treatment of fatty liver and related diseases and applications, as well as the application of TRAF4 inhibitors in the preparation of drugs for preventing, alleviating and / or treating fatty liver and related diseases. Background technique [0002] The liver is the central organ of lipid metabolism in the body, and the fat in the liver mainly comes from food and peripheral adipose tissue. Normal human liver tissue contains a small amount of fat, whose weight is about 3%-5% of the liver weight. When the liver's ability to synthesize fat and / or reduce the ability to transport it into the blood, lipids accumulate too much in the liver , more than 5% of the liver weight or histologically more tha...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K45/00A61K31/7088A61P1/16
CPCA61K31/7088A61K45/00
Inventor 李红良李枫
Owner WUHAN UNIV
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