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Application of capsaicine in inhibition of activation of NLRP3 inflammasome

A technology of inflammasome and capsaicin, applied in the direction of anti-inflammatory agents, organic active ingredients, non-central pain relievers, etc., can solve the problems that have not been reported in research, and achieve the effect of high biological safety

Active Publication Date: 2021-04-30
SOUTH CHINA UNIV OF TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] Capsaicin (Capsaicin, referred to as CAP in this paper) is the active ingredient of pepper, which is often used clinically for analgesia, but so far, no specific research on its inflammatory mechanism has been reported

Method used

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  • Application of capsaicine in inhibition of activation of NLRP3 inflammasome
  • Application of capsaicine in inhibition of activation of NLRP3 inflammasome
  • Application of capsaicine in inhibition of activation of NLRP3 inflammasome

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0050] This example illustrates the inhibitory effect of capsaicin on IL-1β processing, maturation and release downstream of the NLRP3 inflammasome pathway.

[0051] (1) Acquisition of mouse bone marrow-derived macrophages (BMDM): take the bone marrow cells of C57 mice (purchased from Shanghai Nanfang Model Organisms Co., Ltd., SPF grade 8-week-old male mice), and prepare 1×10 6 The density of each / ml was inoculated into a large dish, and the supernatant of L929 cells containing 20% ​​(v / v) (L929 cells were purchased from Shanghai Kebai Company, article number CBP60878, and the medium containing secretions obtained from laboratory cultured L929 cells) and 10% (v / v) fetal bovine serum (purchased from Shanghai Weike Biotechnology Co., Ltd., Cat. No. S1580-500) was cultured in DMEM medium for 5 days, and the DMEM complete medium containing L929 cell supernatant was replaced once every 3 days. .

[0052] (2) In a 24-well plate, according to 5×10 5 BMDM obtained in step (1) was i...

Embodiment 2

[0059] This example is used to illustrate the effect of capsaicin on the production of ROS during the activation of NLRP3 inflammasome.

[0060] (1) Acquisition of BMDM cells: same as step (1) in Example 1.

[0061] (2) In a six-well plate, according to 2×10 6 Cells / well were seeded in BMDM for overnight culture. The next day, add LPS with a final concentration of 100ng / ml for 3h, then add capsaicin with a final concentration of 40μM for 1h, add nigericin with a final concentration of 5μM for 30min, remove the supernatant, digest and dissociate The cell suspension was obtained, and PBS buffer containing ROS fluorescent probe DCFH-DA (purchased from Thermo Fisher, product number C2938) was added, incubated in the dark for 15 min, and the level of ROS in the cells was detected by flow cytometry. The result is as Figure 6 Shown (LPS+Capsaicin: the treatment group that only added LPS and Capsaicin; Nigericin+Capsaicin: the treatment group that added LPS, Nigericin and Capsaici...

Embodiment 3

[0064] This example is used to illustrate the inhibitory effect of capsaicin on ASC oligomerization events during NLRP3 inflammasome activation.

[0065] (1) Acquisition of BMDM: Same as step (1) in Example 1.

[0066] (2) In a six-well plate, according to 2×10 6 Cells / well were seeded in BMDM for overnight culture. The next day, LPS with a final concentration of 100 ng / ml was added for 3 h, then capsaicin (CAP) with a final concentration of 40 μM was added for 1 h, and nigericin (Nigericin) with a final concentration of 5 μM was added for 30 min, and the supernatant was removed. Add NP-40 lysate containing protease inhibitors to lyse the cells, shake and incubate on ice for 30 min. A part of the cell lysate was then taken as an internal reference for the next immunoprecipitation experiment, and the remaining lysate was centrifuged at 6000×g for 10 min at 4°C. The precipitate was taken and washed three times with PBS buffer, and then DSS (Disuccinimidyl suberate, disuccinim...

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Abstract

The invention discloses an application of capsaicine in inhibition of activation of NLRP3 inflammasome. Researches find that capsaicin can inhibit activation of different agonists such as nigericin, microcrystalline sodium urate and alum on NLRP3 inflammasomes, but does not affect activation of inflammasomes such as NLRC4 and AIM2, and can also inhibit assembly of the NLRP3 inflammasomes, including combination of NLRP3 and ASC protein and formation of ASC spots. In addition, the invention also finds that capsaicine can inhibit lipopolysaccharide-induced septicemia. Therefore, the capsaicine can be used as an NLRP3 inflammasome activation inhibitor and a potential medicine for preventing or treating diseases related to abnormal activation of NLRP3 inflammasome.

Description

technical field [0001] The invention belongs to the fields of biology and medicine, and particularly relates to the application of capsaicin in inhibiting the activation of NLRP3 inflammasome. Background technique [0002] The inflammasome is a cytoplasmic multiprotein complex composed of members of the Nod-like receptor (NLR) family and PYHIN (pyrin and HIN domain) family members, which can be activated by various pathogen-associated molecular patterns or injury-associated molecular patterns. The function of the inflammasome is to activate caspase-1, which in turn causes the maturation and secretion of pro-inflammatory cytokines interleukin (IL)-1β and IL-18, and induces pyroptosis. The NLR family protein 3 (NLRP3) inflammasome is a macromolecular multiprotein complex composed of NLRP3, the adapter protein ASC and the effector protein Caspase-1. Unlike other inflammasomes, the NLRP3 inflammasome can be activated by a variety of stimuli, including microbial components and e...

Claims

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Application Information

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IPC IPC(8): A61K31/165A61P29/00
CPCA61K31/165A61P29/00
Inventor 张云娇邓育辉温龙平
Owner SOUTH CHINA UNIV OF TECH
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