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Application of histone demethylase KDM5C (lysine(K) demethylase 5C) in screening of drugs for treating the fatty liver and related diseases of the fatty liver

A technology for demethylating enzymes and histones, applied in the field of gene function and application, can solve problems such as unclear functions, and achieve the effect of worsening fatty liver disease

Active Publication Date: 2018-06-29
武汉惠康达科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the role of KDM5C in hepatocytes is still unclear

Method used

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  • Application of histone demethylase KDM5C (lysine(K) demethylase 5C) in screening of drugs for treating the fatty liver and related diseases of the fatty liver
  • Application of histone demethylase KDM5C (lysine(K) demethylase 5C) in screening of drugs for treating the fatty liver and related diseases of the fatty liver
  • Application of histone demethylase KDM5C (lysine(K) demethylase 5C) in screening of drugs for treating the fatty liver and related diseases of the fatty liver

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0092] [Example 1] Establishing a KDM5C overexpressed L02 stable transfection cell line

[0093] According to the steps for establishing a stable transgenic cell line with overexpression of L02 in the embodiment, a stable transgenic cell line of L02 with overexpression of KDM5C was established. Afterwards, the cells were collected, and the expression of KDM5C was verified by WB. WB results such as figure 1As shown, compared with the control cells, the protein expression of KDM5C in KDM5C-TG L02 cells was significantly increased, indicating that the construction of stable transfected cell lines is effective.

Embodiment 2

[0094] [Example 2] Effect of KDM5C knockdown on fat accumulation in liver cells

[0095] 1. Grouping of experimental cells: Normal L02 cells and KDM5C-KO L02 cells were divided into 2 groups, namely normal L02 cell control group, KDM5C-KO L02 cell control group, normal L02 cell experimental group, and KDM5C-KOL02 cell experimental group.

[0096] 2. Establishment and detection of fatty liver cell model: when the cells adhere to the wall and the cell density is about 30%, add palmitate (palmitate, PA) and oleic acid (oleic acid, OA) ( PA 0.2mM+OA 0.4mM) were stimulated, and 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.

[0097] The results of Oil Red O staining were as follows: figure 2 As shown, the cells in the control group had no obvious red fat droplets, but when stimulated by adding PA+OA, the area of ​​cells stained red by Oil Red O was significantly increased compared with the...

Embodiment 3

[0098] [Example 3] Effect of KDM5C Gene Overexpression on Fat Accumulation in Hepatocytes

[0099] 1. Experimental cell grouping: Vector L02 cells and KDM5C-TG L02 cells were divided into 2 groups: VectorL02 cell control group, KDM5C-TG L02 cell control group, Vector L02 cell experimental group, and KDM5C-TG L02 cell experimental group.

[0100] 2. Establishment and detection of fatty liver cell model: when the cells adhered to the wall and the cell density reached about 30%, PA and oleic acid OA (PA 0.2mM+OA 0.4mM) were added to the two experimental groups for stimulation, and the control The same amount of BSA was added to the group, and Oil Red O staining was carried out after 12 hours. The staining procedure was as described in Example 2.

[0101] The results of Oil Red O staining were as follows: image 3 As shown, the cells in the control group had no obvious red fat droplets, but when stimulated by adding PA+OA, the fat droplets stained red by Oil Red O increased signi...

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Abstract

The invention discloses an application of histone demethylase KDM5C (lysine(K) demethylase 5C) in screening of drugs for treating the fatty liver and related diseases of the fatty liver. A human normal hepatocyte L02 cell line, an L02 cell line with overexpressed histone demethylase KDM5C and an L02 cell line with KDM5C (CRISPR-Cas9) knockout are adopted as experimental subjects, and the functionof the gene KDM5C is studied by a fatty liver cell model induced by palmitic acid and oleic acid jointly. Studies show that the accumulation of lipid in L02 cells induced by PA and OA is significantlyincreased during overexpression of KDM5C; the accumulation of lipid in the L02 cells induced by PA and OA is significantly decreased when expression of KDM5C is decreased, that is, expression of thegene KDM5C can promote generation and development of the fatty liver and related diseases. Based on the function of KDM5C in deterioration of the fatty liver, a target for development of drugs for prevention, alleviation and / or treatment of the fatty liver and related diseases is provided.

Description

technical field [0001] The invention belongs to the field of gene function and application, in particular to the application of a histone demethylase (lysine (K) demethylase 5C, KDM5C) as a drug target in screening drugs for the treatment of fatty liver, and the use of KDM5C inhibitors in Application in the preparation of medicines for treating fatty liver. Background technique [0002] With the improvement of residents' living standards, the incidence of fatty liver is also increasing year by year. Fatty liver is a clinicopathological syndrome characterized by excessive fat deposition in liver cells. Clinically, patients can be divided into alcoholic fatty liver disease and nonalcoholic fatty liver disease according to whether they have a history of excessive drinking, and the proportion of patients with alcoholic fatty liver disease is 5%. The patients with nonalcoholic fatty liver disease account for the vast majority And showing an upward trend year by year. Nonalcohol...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6883A61K45/00A61K49/00A61P1/16
CPCA61K45/00A61K49/0008C12Q1/6883C12Q2600/158
Inventor 李红良李枫
Owner 武汉惠康达科技有限公司
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