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Application of deubiquitinase 1 containing otu functional domain in the preparation of drugs for the treatment of fatty liver and related diseases

A technology for deubiquitinating enzymes and functional domains, used in the preparation of drugs for preventing, relieving and/or treating fatty liver, in the field of deubiquitinating enzymes, and achieving high safety effects

Active Publication Date: 2020-06-09
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 OTUD1 in fatty liver in the present invention

Method used

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  • Application of deubiquitinase 1 containing otu functional domain in the preparation of drugs for the treatment of fatty liver and related diseases
  • Application of deubiquitinase 1 containing otu functional domain in the preparation of drugs for the treatment of fatty liver and related diseases
  • Application of deubiquitinase 1 containing otu functional domain in the preparation of drugs for the treatment of fatty liver and related diseases

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0101] [Example 1] Establishment of L02 cell line stably transfected with OTUD1

[0102] According to the steps of establishing the L02 stable transfection cell line in the embodiment, the L02 stable transfection cell line with OTUD1 overexpression and knockdown was established. Afterwards, the cells were collected, and the expression of OTUD1 was verified by WB. The result is as figure 1 As shown, in L02 cells infected with OTUD1 overexpression lentivirus system, the expression of OTUD1 was significantly increased; in L02 cells infected with OTUD1 knockdown lentivirus system, the expression of OTUD1 was significantly decreased, indicating that the cell line was successfully established.

Embodiment 2

[0103] [Example 2] Effect of OTUD1 knockdown on fat accumulation in liver cells

[0104] (1) Grouping of experimental cells: normal L02 cell control group, OTUD1 stable knockdown L02 cell control group, normal L02 cell experimental group, and OTUD1 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, but the area of ​​red fat droplets in the cells in the experimental group was significantly increased after adding PA+OA stimulation, and the area of ​​red fat droplets in the OTUD1 stably ...

Embodiment 3

[0107] [Example 3] Effect of OTUD1 overexpression on fat accumulation in liver cells

[0108] 1. Experimental cell grouping: normal L02 cell control group, OTUD1 stable overexpression L02 cell control group, normal L02 cell experimental group, OTUD1 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 have no obvious red color, and when the experimental group is stimulated by adding PA+OA, the cells in the area of ​​red fat droplets in the experimental group are significantly increased com...

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Abstract

The invention discloses an application of OTU Domain-Containing Deubiquitinase 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 OTUD1 knockout and overexpression vector system, and an OTUD1 gene knockout or overexpressed L02 cellis obtained. The function of the OTUD1 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 OTUD1 gene, the red fat droplets in the L02 cell knockouted by the OTUD1 gene are large and multiple; and on the contrary, and the red fat droplets in the L02 cell overexpressed by the OTUD1 gene are small and less. The result shows that the OTUD1 gene can obviously inhibit liver lipid deposition and inhibit generation of fatty liver.

Description

technical field [0001] The present invention belongs to the field of gene function and application, and particularly relates to the function and application of a deubiquitinase, that is, deubiquitinase 1 (OTU Deubiquitinase 1, OTUD1) containing an OTU functional domain, in non-alcoholic fatty liver. And the application of OTUD1 as a target gene in the preparation of drugs for preventing, alleviating and / or treating fatty liver. Background technique [0002] With the development of the economy and the improvement of living standards, the incidence of non-alcoholic fatty liver disease (NAFLD) is increasing year by year, and it has become one of the most common liver diseases in the world. NAFLD is a clinicopathological syndrome characterized by hepatic steatosis and lipid storage in the absence of a history of excessive alcohol consumption [1][2] According to the pathological development process, it can be divided into simple non-alcoholic fatty liver, non-alcoholic steatohep...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K38/48A61K45/00A61K48/00G01N33/573A61P1/16A61P5/50A61P3/00A61P3/04A61P3/10A61P3/06A61P35/00
CPCA61K38/4813A61K45/00A61K48/005C12Y304/19012G01N33/573G01N2333/948G01N2500/00
Inventor 李红良
Owner WUHAN UNIV
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