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Application of otub2 and its inhibitors in the preparation of drugs for the treatment of fatty liver and related diseases

A technology of OTUB2 and fatty liver, which is applied in the field of gene function and application, and can solve problems such as transient activation, reduced cell sensitivity, and consumption

Active Publication Date: 2020-08-25
WUHAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In addition, studies have also shown that depletion of OTUB2 can also transiently activate the G2 checkpoint, thereby reducing the sensitivity of cells to aging
However, the role of OTUB2 in fatty liver has not yet been reported

Method used

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  • Application of otub2 and its inhibitors in the preparation of drugs for the treatment of fatty liver and related diseases
  • Application of otub2 and its inhibitors in the preparation of drugs for the treatment of fatty liver and related diseases
  • Application of otub2 and its inhibitors in the preparation of drugs for the treatment of fatty liver and related diseases

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0109] [Example 1] Construction and verification of stable transfected cell lines L02-phage and L02-OTUB2-phage-3×flag

[0110] Build verification results such as figure 1 : The size of the endogenous OTUB2 is 27KDa. It was detected by both the body antibody and the tag antibody. Compared with the vector control group, it can be clearly seen that the overexpression cell line was successfully constructed.

Embodiment 2

[0111] [Example 2] Effect of overexpression of OTUB2 on fat accumulation in L02 cells

[0112] Cell treatment: After the overexpression OTUB2 cell line and the phage cells in the control group adhered to the wall, they were starved for 12 hours, then treated with 0.25 / 0.5mM PA / OA for 24 hours, and then stained with oil red.

[0113] The staining result is as figure 2 A, 2B: Compared with the phage control group, the staining of the overexpressed OTUB2 cell line was significantly deeper. The results indicate that overexpression of OTUB2 can significantly promote fat accumulation.

Embodiment 3

[0114] [Example 3] L02-siRNA transfection and western blot detection

[0115] Cell treatment: L02 cells were divided into 5 groups, one group was blank cells, and the other four groups were transfected with siNC control, siOTUB2#S1, siOTUB2#S2, and siOTUB2#S1+S2. After the cells adhered to the wall and grew to a transfectable density, they were transfected with siRNA, and then transfected again after 24 hours. The samples were collected 48 hours after transfection, and then Western Blot was used to detect the knockdown effect.

[0116] The result is as image 3 : Compared with L02 blank cells and siNC control group, the expression of OTUB2 in the experimental group was significantly decreased. This result indicates that siOTUB2 is effective.

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Abstract

The invention discloses application of OTUB2 and an OTUB2 inhibitor to preparing a medicine for treating fatty liver and related diseases. An LO2 cell is taken as the experimental subject to respectively construct OTUB2 overexpressed and OTUB2 knock-down LO2 cell models, and oil red O staining is conducted after the simulation by palmitic acid / oleic acid (PA / OA). The staining result shows that: compared with a phage control group, the staining of an OTUB2 overexpression group is deepened, and the staining of an OTUB2 knock-down group becomes shallow. The result shows that the fat accumulationis promoted and the fatty liver is aggravated after the overexpression of an OTUB2 gene; the fat accumulation is inhibited after the OTUB2 gene is knocked out, and the occurrence of the fatty liver isimproved. Therefore, the OTUB2 provides a target for the preparation of the medicine used for preventing, relieving and / or treating the fatty liver and the related diseases.

Description

technical field [0001] The invention belongs to the field of gene function and application, and particularly relates to the application of a deubiquitinating enzyme OTUB2 as a drug target in the screening of drugs for the treatment of fatty liver, and the use of OTUB2 inhibitors in the preparation of prevention, alleviation and / or treatment of fatty liver and Drug application in related diseases. Background technique [0002] With the continuous improvement of people's living standards, great changes have taken place in the diet structure, which gradually tends to be high in protein and high in fat. The liver occupies an important position in the body's fat metabolism, and it participates in many important links in the lipid metabolism process, including the intake and synthesis of fatty acids, the processing, storage, oxidative decomposition and output of lipids. When the amount of fatty acid obtained by the liver exceeds its processing capacity, lipids are deposited in th...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K45/00A61P1/16A61P35/00
CPCA61K45/00
Inventor 李红良李枫
Owner WUHAN UNIV
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