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Application of nuclear factor kappa b inhibitory protein 3 combined with a20 in the preparation of drugs for treating fatty liver and related diseases
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A technology to inhibit protein and nuclear factor, applied in gene therapy, metabolic disease, drug combination, etc., to achieve high safety effect
Active Publication Date: 2020-06-09
WUHAN UNIV
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Problems solved by technology
But there is no report about the function and application of ABIN3 of the present invention in fatty liver disease
Method used
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Embodiment 1
[0097] [Example 1] Effect of ABIN3 overexpression on fat accumulation in L02 cells
[0098] Construct a lentiviral expression vector that overexpresses ABIN3, transfect HEK-293T cells, package the lentivirus, infect L02 cells to construct a stable cell line that overexpresses ABIN3, and overexpress the empty vector as a control, and detect whether the stable cell line expresses it by Western blot ABIN3.
[0099] After the detection, the cells were divided into 4 groups, L02 empty control group, ABIN3 overexpressed L02 stable cell line control group, L02-empty experimental group, ABIN3 overexpressed L02 stable cell line experimental group. After the cells adhered to the wall and the cells were cultured to 50% healing, palmitate (PA) and oleic acid (OA) (PA 0.2mM+OA 0.4mM) were added to the two experimental groups for stimulation, and the same amount was added to the control group. After 12 hours, the cell samples of each group were collected for Oil Red O staining experiment. ...
Embodiment 2
[0101] [Example 2] Effect of ABIN3 overexpression on fat accumulation in primary mouse liver cells
[0102] Construct a lentiviral expression vector that overexpresses ABIN3, transfect HEK-293T cells, package the lentivirus, and infect mouse primary liver cells to construct a cell model that overexpresses ABIN3, while overexpressing the empty vector as a control (Con), through Western Blot was used to detect whether ABIN3 was overexpressed in ABIN3-infected cells.
[0103] After the detection was completed, the cells were divided into 4 groups, namely: no-load control group, ABIN3 overexpressed cell control group, L02-no-load experimental group, and ABIN3 overexpressed cell experimental group. After the cells adhered to the wall and the cells were plated to a degree of 50% healing, palmitate (PA) and oleic acid (OA) (PA 0.2mM+OA 0.4mM) were added to the two experimental groups for stimulation, and the same amount was added to the control group. After 12 hours, the cell sample...
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Abstract
The invention discloses application of nuclear factor kappa B arrestin 3 combined with A20 in preparation of drugs for treating fatty liver and related diseases, and belongs to new use of ABIN3. An L02 cell line, an ABIN3 over-expressing L02 cell line, a mouse primary hepatocyte cell, and an ABIN3 over-expressing mouse primary hepatocyte cell are used as an experimental object, and a fatty liver cell model is induced through combination of palmitic acid and oleic acid. Oil red O staining results show that compared with L02 cells, an area of red fat droplets in ABIN3 over-expressing L02 cells is significantly reduced under combined stimulation of the palmitic acid and the oleic acid at the same concentration. A same trend of the oil red O staining results is also observed in the ABIN3 over-expressing mouse primary hepatocyte cell, indicating that ABIN3 overexpression significantly inhibits lipid accumulation in fatty liver cells induced by the palmitic acid and the oleic acid. Therefore, ABIN3 has the functions of inhibiting liver lipid accumulation and protecting liver.
Description
technical field [0001] The present invention belongs to the field of gene function and application, and in particular relates to a ubiquitin chain binding protein, that is, nuclear factor κB inhibitory protein 3 (A20binding inhibitor of NF-κB activation, ABIN3) combined with A20 in fatty liver and related diseases Function and application, and the application of ABIN3 as a target gene in the preparation of drugs for preventing, alleviating and / or treating fatty liver and related diseases. Background technique [0002] Non-alcoholic fatty liver disease (NAFLD) is a chronic liver disease characterized by fatty degeneration and fat accumulation in hepatic parenchymal cells despite no history of excessive alcohol consumption[1]. Studies have shown that about 10% of NAFLD patients will develop non-alcoholic steatohepatitis (NASH), and about 20% of these NASH patients will develop liver cirrhosis within 10 years. According to reports, the incidence of NAFLD in developed countries...
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