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Application of piperine to preparing medicines for regulating AMPK immunometabolism pathways of immune cells

A technology for regulating immunity and piperine, applied in the field of medicine, can solve problems that have not been reported before, and achieve the effect of repairing cellular immune metabolism and preventing functional exhaustion.

Inactive Publication Date: 2017-01-04
JINAN UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0008] The application of piperine as a drug for regulating the AMPK signaling pathway has not yet been reported

Method used

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  • Application of piperine to preparing medicines for regulating AMPK immunometabolism pathways of immune cells
  • Application of piperine to preparing medicines for regulating AMPK immunometabolism pathways of immune cells
  • Application of piperine to preparing medicines for regulating AMPK immunometabolism pathways of immune cells

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0034] (1) Cell culture and treatment

[0035] Mouse macrophages J774A.1 were cultured in complete DMEM medium. After the cells grew to the logarithmic growth phase, they were planted in 6-well plates with a cell density of 2.5×10 5 / well (2mL culture medium), cultivated for 24 hours. The cells were pretreated with different concentrations of piperine (20, 40, 80 μmol / L) for 4 hours, then 20 μl of 50 μg / ml LPS was added to make the final concentration 500 ng / ml, and the cells were activated for 4 hours. After washing with DMEM medium containing 0.1% FBS, the cell culture medium was replaced with DMEM medium (1.2 mL) containing 0.1% FBS and 4 mmol / L ATP for 1 hour.

[0036] (2) Detection method of AMPK activity:

[0037]After the cells were treated as above, the cells were lysed with 2×SDS-loading buffer (2×SDS-loading buffer), and the total protein level of AMPK in the cells was detected by Western blotting (detected with AMPK antibody ) and phosphorylated AMPK protein lev...

Embodiment 2

[0042] (1) Cell culture and drug treatment:

[0043] The mouse mononuclear macrophage J774A.1 cell culture method is the same as "Example 1". After the cells grew to the logarithmic growth phase, they were planted in 6-well plates with DMEM complete medium, and the cell density was 2.5×10 5 / well (2mL medium), pretreated with DMEM complete medium containing 80μmol / L piperine for 4 hours, added 50μg / ml LPS 20μl to make the final concentration 500ng / ml, and treated for 4 hours to activate the cells. Then, after washing with DMEM medium containing 0.1% FBS, the cell culture solution was replaced with DMEM medium (1.2 ml) containing 0.1% FBS and 1 mmol / L metformin (Metformin, AMPK activator) for 1 hour. Then add 48 μl of ATP solution with a concentration of 100 mmol / L to make the final concentration 4 mmol / L, and treat with ATP for 1 hour.

[0044] (2) Detection method of AMPK activity:

[0045] Same as "Example 1".

[0046] (3) Experimental results

[0047] image 3 It is t...

Embodiment 3

[0049] (1) Cell culture and treatment

[0050] Mouse fibroblast L929 cells were purchased from the Cell Bank of Kunming Institute of Zoology, Chinese Academy of Sciences. The cell culture method refers to the operation procedure of Monack laboratory (Stanford), and 1 × 10 8 Cells are cultured on a bottom area of ​​500 cm 2 In the three-layer culture flask, the medium is DMEM complete medium, 200mL, and cultivated for one week.

[0051] The culture method of mouse bone marrow-derived macrophages (BMDM): C57BL / 6 mice, 6-8 weeks old, female, were purchased from the Experimental Animal Center of Southern Medical University. After being sacrificed by cervical dislocation, the femur was taken, and the bone marrow cells were washed out with DMEM medium with a syringe, and centrifuged at 300×g. The obtained cells were cultured in a bacterial culture dish (density of 5 × 10 6 / dish, medium volume is 8-10mL). After culturing for 3 days, add 10 mL of new BM-Mac medium; on the 5th da...

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Abstract

The invention belongs to the field of medicines, and discloses application of piperine to preparing medicines for regulating AMPK immunometabolism pathways of immune cells. The piperine is used as an active constituent in the medicines for regulating the AMPK immunometabolism pathways of the immune cells. AMPK metabolism signal pathways of the cells can be changed when the immune cells are stimulated by inflammation such as bacterial infection, AMPK metabolism signal balance can be broken under stimulation effects of second signals of ATP (adenosine triphosphate) and the like, the activity can be subjected to sharp up-regulation, but the AMPK activity of the cells treated by the piperine can be inhibited, and the cells can be prevented from pyroptosis death. The application has the advantages that the piperine has definite AMPK immunometabolism pathway regulation activity and has important significance on preventing functional depletion of the cells in the aspect of cell immunometabolism repair.

Description

technical field [0001] The invention belongs to the field of medicines, and in particular relates to the application of piperine as an immunoregulatory medicine in the preparation of medicines for regulating the AMPK immune metabolism pathway of immune cells. Background technique [0002] AMPK is a key molecule that regulates energy metabolism. It not only regulates immune metabolism (immunometabolism), but also regulates cell survival and affects a series of cellular metabolic activities. For example, AMPK activates the key kinase of lipid metabolism ACC through phosphorylation, and inhibits lipid synthesis; AMPK can also inhibit the activity of mTORC1 by phosphorylating Raptor (the constituent protein of mTORC1 complex) or TSC2 (mTORC1 inhibitor), thereby limiting the protein synthesis and cell proliferation. [0003] Microbial infection, high-fat diet, etc. can change the energy metabolism of innate immune cells, which in turn affects the function of innate immune cells....

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K31/4525A61P37/02
CPCA61K31/4525
Inventor 何贤辉欧阳东云梁译丹徐丽慧李陈广魏红霞潘浩
Owner JINAN UNIVERSITY
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