Application of berberine in preparation of medicines for enhancing inflammasome activation

A technology of inflammasomes and berberine, which is applied in the fields of biomedicine and medicine, can solve problems such as increasing risks, and achieve the effects of enhancing function, enhancing inflammasome activation, and enhancing the ability to kill intracellular bacteria

Inactive Publication Date: 2016-12-07
JINAN UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Third, pyroptosis is one of the important exit mechanisms for activated macrophages: if macrophages continue to be activated without pyroptosis, their immunometabolic pathways have been altered, leading to lipid deposition and becoming foam cells or adipose macrophages cells, which continuously secrete inflammatory factors (endogenous pyrogens) and chemokines, recruit other immune cells to the focal tissue, and increase the risk of chronic inflammatory diseases such as atherosclerosis and obesity
[0007] Currently, there is no report on the function of berberine in enhancing inflammasome activation

Method used

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  • Application of berberine in preparation of medicines for enhancing inflammasome activation
  • Application of berberine in preparation of medicines for enhancing inflammasome activation
  • Application of berberine in preparation of medicines for enhancing inflammasome activation

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029] (1) Cell culture and treatment

[0030] Mouse macrophage J774A.1 was purchased from the Shanghai Cell Bank of the Chinese Academy of Sciences and cultured in DMEM (complete medium) containing 10% (v / v) fetal bovine serum and 100 U / mL penicillin and 100 μg / mL streptomycin . After the cells grew to the logarithmic growth phase, they were seeded in 6-well plates with a cell density of 2.5×10 5 / well (2mL culture medium), cultivated for 24 hours. Then change the medium, add complete medium (2mL) containing 500ng / mL LPS, pretreat for 4 hours, activate J774A. (v / v) FBS DMEM medium (1.2 mL) was treated for 1 hour, and finally 0.8 mL of ATP-containing DMEM medium (ATP final concentration was 3 mmol / L) was added for 1 hour.

[0031] (2) Detection method of caspase-1 and HMGB1 in the supernatant

[0032] After the above-mentioned treatment of the cells, absorb the culture supernatant, centrifuge at 200×g for 5 minutes, transfer the supernatant to another centrifuge tube, and ...

Embodiment 2

[0036] (1) Cell culture and treatment

[0037]C57BL / 6 mice, female, 6-8 weeks old, were purchased from the Experimental Animal Center of Southern Medical University. After one week of adaptive breeding, they were stimulated by intraperitoneal injection of 1 mL of PBS solution containing 3% (w / v) thioglycollate (TG) for 4 days, and sacrificed by cervical dislocation, and the free cells in the peritoneal cavity were washed out with PBS solution. Cultured in 6-well plates containing 10% FBS and DMEM complete medium containing 100 U / mL penicillin and 100 μg / mL streptomycin, the cell density was 2.5×10 6 / well (2mL culture medium), cultivated for 24 hours. Then change the medium, add complete medium (2mL) containing 500ng / mL LPS to pretreat for 4 hours, activate peritoneal macrophages, and then pass through medium containing different concentrations of berberine (0.75, 1.5, 3.0μmol / L) (1.2 mL) was treated for 1 hour, and finally ATP was added (to make the final concentration 2 mm...

Embodiment 3

[0043] (1) Cell culture and treatment

[0044] 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 culture medium is DMEM complete medium (200mL).

[0045] 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 cells / dish, medium volume 8-10mL). After culturing for 3 days, add 10 mL of new medium; on the 5th day of culturing, replace with the above-mentioned f...

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Abstract

The invention discloses application of berberine in preparation of medicines for enhancing inflammasome activation. The inventor discovers that berberine can achieve the effects of obviously promoting inflammasome activation induced by signals such as ATP and the like, improving activity of caspase-1 and secretion of mature IL-1beta, further promoting bacteria killing effects of macrophage and obviously increasing survival rate of mice in an intra-abdominal bacterial infection model, so that the berberine can enhance functions of innate immunocytes such as macrophage and the like and obviously improving resistance of the host to bacterial infection by promoting secretion of IL-1beta. Therefore, the berberine can be used for preparing inflammasome activation enhanced drugs. The drug can achieve the effects of enhancing functions of the innate immunocytes and treating systemic bacterial infection and other related diseases.

Description

technical field [0001] The invention belongs to the field of biomedicine and medicine, and specifically relates to the application of berberine in the preparation of drugs for enhancing the activation of inflammasomes. Background technique [0002] Berberine, also known as berberine, is a common isoquinoline alkaloid with molecular formula C 20 h 18 NO 4 . It exists in many plants of four families and ten genera including Berberidaceae, and can be extracted from plants such as Coptis chinensis, Cortex Phellodendri, and three needles. Berberine is a yellow needle-like crystal with a melting point of 145°C. It is insoluble in water but soluble in benzene, ether and chloroform. The commonly used drug form is berberine hydrochloride (the structural formula is shown below), and its solubility in water is relatively small. Berberine has inhibitory effects on Shigella, Mycobacterium tuberculosis, Pneumococcus, Typhoid bacillus and other bacteria, among which the effect on Shig...

Claims

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Application Information

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IPC IPC(8): A61K31/4375A61P29/00A61P31/04A61P37/04
CPCA61K31/4375
Inventor 何贤辉欧阳东云徐丽慧李陈广潘浩梁译丹魏红霞
Owner JINAN UNIVERSITY
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