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Application of piperine in preparation of drugs for resisting pyroptosis and multiple organ injury

A multi-organ, piperine technology, applied in antibacterial drugs, drug combinations, antidote, etc., to prevent damage to multiple organs

Inactive Publication Date: 2016-10-26
JINAN UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] The effect of piperine on pyroptosis has not been reported yet

Method used

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  • Application of piperine in preparation of drugs for resisting pyroptosis and multiple organ injury
  • Application of piperine in preparation of drugs for resisting pyroptosis and multiple organ injury
  • Application of piperine in preparation of drugs for resisting pyroptosis and multiple organ injury

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] (1) Cell culture and treatment

[0027] Mouse mononuclear macrophage J774A.1 cells were purchased from the Shanghai Cell Bank of the Chinese Academy of Sciences and cultured in DMEM containing 10% fetal bovine serum and 100 U / ml penicillin (penicillin), 100 μg / ml streptomycin (streptomycin) ( complete medium). After the cells grow to the logarithmic growth phase, they are planted in a 24-well plate with a cell density of 3×10 4 / well (500μl culture medium), cultivated for 24h. Then (change the medium) add piperine of different concentrations (1, 5, 10, 20, 40, 80 μmol / L) each 500 μl, pretreatment for 24 hours. Then add 50 μg / ml LPS 5 μl to make the final concentration 500ng / ml, and treat for 4 hours to activate the cells. Finally, the cell culture medium was replaced with a complete medium containing 0.1% FCS and 5mmol / L ATP, the medium volume was 500 μl, and the ATP treatment time was 1 h.

[0028] (2) Detection and observation of pyroptotic cells

[0029] After t...

Embodiment 2

[0033] (1) Cell culture

[0034] L929 cells were purchased from the Cell Bank of Kunming Institute of Zoology, Chinese Academy of Sciences. The cell culture method refers to Monack Labprotocol (Stanford), and 1×10 8 Cells are cultured on a bottom area of ​​500 cm 2 In the Erlenmeyer flask, the culture medium is DMEM complete medium, 200ml.

[0035] 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 flushed out with DMEM medium with a syringe. 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 medium; on the 5th day of culturing, replace with the above-mentioned fresh medium. On the 7th day of culture, the cells were collected and ...

Embodiment 3

[0043] Mouse macrophage J774A.1 cells were planted in a 6cm-diameter petri dish with DMEM complete medium. After the cells entered the logarithmic growth phase, they were treated with DMEM Complete medium (4ml / well) was pretreated for 24h; then add 40μl LPS solution (working solution concentration 50μg / ml to make the final concentration reach 500ng / ml) for 4h; finally replace the medium with 0.1% FCS and 4mmol / well L ATP in DMEM medium (4ml) was treated for 1h. After the cells were treated as above, the expression of pyroptosis-related proteins or signaling molecules was detected by immunoblotting. from Figure 4 It can be seen that after the cells are treated with LPS+ATP (the third lane in the figure), caspase-1 is activated (Figure A, supernatant, pro-caspase-1 is processed into the active form of caspase-1), and pro-IL-1β is up-regulated The expression of PIP pretreated cells can inhibit the activation of caspase-1 up-regulated by LPS+ATP and the release of HMGB1 (a dang...

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Abstract

The invention discloses application of piperine in preparation of drugs for resisting pyroptosis and multiple organ injury. The invention is provided on the basis that an inventor of the invention firstly discovers that the piperine has effect of resisting pyroptosis and multiple organ injury. The inventor of the invention discovers that the piperine can remarkably inhibit pyroptosis induced by bacterial infection, release of LPS+ATP induced proinflammatory factors IL-1 beta and dangerous signal molecules HMGB1 is inhibited remarkably, and multiple organ injury caused by bacterial infection can be prevented effectively. At present, specific drugs which can effectively prevent pyroptosis and multiple organ injury which are induced by bacterial infection and fungal infection do not exist clinically. The application of the piperine in preparation of the drugs for preventing pyroptosis and multiple organ injury which are caused by infection has significance on clinical treatment of septicemia and the like, and can be used for developing associated drugs.

Description

technical field [0001] The invention belongs to the field of medicines, and in particular relates to the application of piperine in the preparation of medicines for resisting pyroptosis and multiple organ damage. Background technique [0002] Pyroptosis is a programmed cell death mode activated by inflammation. Specifically, infections such as bacteria and fungi can cause damage to multiple organs such as the liver, leading to septic shock. There are many reasons for multi-organ injury. The latest research shows that after microbial infection, human cells (including immune cells) recognize pathogen-associated molecular patterns (such as bacterial LPS) with the help of their pattern recognition receptors, and the cells are activated accordingly to initiate NF-κB and inflammasome signaling pathways , Promote the expression of pro-IL-1β and other inflammatory factors (precursors). Under the stimulation of the second signal such as ATP, the above-mentioned activated cells can...

Claims

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Application Information

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IPC IPC(8): A61K31/4525A61P29/00A61P31/04A61P31/10A61P39/00
CPCA61K31/4525
Inventor 欧阳东云何贤辉梁译丹徐丽慧李陈广魏红霞潘浩
Owner JINAN UNIVERSITY
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