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Application of ubiquitin aldehyde-binding protein 1 containing otu functional domain in the preparation of drugs for treating fatty liver and related diseases

A technology that combines proteins and functional domains, and is used in medical preparations containing active ingredients, metabolic diseases, drug combinations, etc., to achieve the effect of protecting fatty liver

Active Publication Date: 2020-12-01
WUHAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

But there is no report about the function and application of OTUB1 of the present invention in NAFLD

Method used

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  • Application of ubiquitin aldehyde-binding protein 1 containing otu functional domain in the preparation of drugs for treating fatty liver and related diseases
  • Application of ubiquitin aldehyde-binding protein 1 containing otu functional domain in the preparation of drugs for treating fatty liver and related diseases
  • Application of ubiquitin aldehyde-binding protein 1 containing otu functional domain in the preparation of drugs for treating fatty liver and related diseases

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0101] [Example 1] Establishment of OTUB1 stably transfected L02 cell line

[0102] According to the steps of establishing the L02 stable transfection cell line in the embodiment, the L02 stable transfection cell line with OTUB1 overexpression and knockdown was established. Afterwards, the cells were collected, and the expression of OTUB1 was verified by WB. The result is as figure 1 As shown, it can be seen that the expression of OTUB1 was significantly increased in the L02 cells infected with the OTUB1 overexpression lentivirus system; the expression of OTUB1 was significantly decreased in the L02 cells infected with the OTUB1 knockdown lentivirus system, indicating that the cell line was successfully established.

Embodiment 2

[0103] [Example 2] Effect of OTUB1 knockdown on fat accumulation in liver cells

[0104] (1) Grouping of experimental cells: normal L02 cell control group, OTUB1 stable knockdown L02 cell control group, normal L02 cell experimental group, and OTUB1 stable knockdown L02 cell experimental group.

[0105] (2) Establishment and detection of fatty liver cell model: as soon as the cells adhere to the wall, after the cells are cultured to 50% healing, add palmitate (PA) and oleic acid (OA) (PA 0.2mM +OA 0.4mM) for stimulation, the same amount of BSA was added to the control group, and the cell samples of each group were collected after 12h for Oil Red O staining.

[0106] The results of Oil Red O staining were as follows: figure 2 As shown, the cells in the control group had no obvious red color, and the area of ​​red fat droplets in the cells in the experimental group was significantly increased after adding PA+OA stimulation, and the increase in the area of ​​red fat droplets in ...

Embodiment 3

[0107] [Example 3] Effect of OTUB1 overexpression on fat accumulation in liver cells

[0108] 1. Experimental cell grouping: normal L02 cell control group, OTUB1 stable overexpression L02 cell control group, normal L02 cell experimental group, OTUB1 stable overexpression L02 cell experimental group.

[0109] 2. Establishment and detection of fatty liver cell model: After the cells adhere to the wall and the cells are cultured to 50% healing, add palmitate (PA) and oleic acid (OA) (PA 0.2mM+OA) to the two experimental groups 0.4mM) stimulation, the same amount of BSA was added to the control group, and the cell samples of each group were collected after 12h for Oil Red O staining.

[0110] The results of Oil Red O staining were as follows: image 3 As shown, the cells in the control group had no obvious red color, but when the experimental group was stimulated by adding PA+OA, the cells with the area of ​​red fat droplets in the experimental group were significantly increased ...

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Abstract

The invention discloses an application of OTU Domain-Containing Ubiquitin Aldehyde-Binding Protein 1 in preparation of medicines for treating fatty liver and related diseases. According to the invention, a normal human hepatocyte L02 cell line is taken as a research object, lentivirus can be used for constructing an OTUB1 knockout and overexpression vector system, and an OTUB1 gene knockout or overexpressed L02 cell is obtained. The function of the OTUB1 gene in fatty degeneration is researched through a palmitic acid (PA) and oleic acid (OA) combination-induced fatty liver cell model. The result displays that under same PA+OA stimulation, compared with the L02 cell normally expressed by the OTUB1 gene, the red fat droplets in the L02 cell knockouted by the OTUB1 gene are large and multiple; on the contrary, and the red fat droplets in the L02 cell overexpressed by the OTUB1 gene are small and less. The result shows that the OTUB1 gene can obviously inhibit liver lipid deposition and inhibit generation of fatty liver.

Description

technical field [0001] The invention belongs to the field of gene function and application, in particular to a deubiquitinating enzyme, that is, OTU Domain-Containing Ubiquitin Aldehyde-Binding Protein 1 (OTUB1) in fatty liver The function and application of OTUB1, and the application of OTUB1 as a target gene in the preparation of drugs for preventing, alleviating and / or treating fatty liver. Background technique [0002] Non-alcoholic fatty liver disease (NAFLD) is a clinicopathological syndrome characterized by fatty degeneration and lipid accumulation in hepatocytes without a history of excessive alcohol consumption[1][2]. With the development of human society and economy and the improvement of medical level, the incidence of various chronic liver diseases has remained stable or even decreased, while the incidence of NAFLD has shown a rapid upward trend. The latest epidemiological survey shows that NAFLD is the leading disease of chronic liver disease worldwide, with an...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K38/17A61K48/00A61P1/16A61P5/50A61P3/00A61P3/04A61P3/10A61P3/06A61P35/00G01N33/68
CPCA61K38/1709A61K48/005G01N33/68G01N2333/435G01N2500/00
Inventor 李红良李枫
Owner WUHAN UNIV
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