Application of capsaicin in inhibiting nlrp3 inflammasome activation

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

Active Publication Date: 2022-05-24
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 capsaicin in inhibiting nlrp3 inflammasome activation
  • Application of capsaicin in inhibiting nlrp3 inflammasome activation
  • Application of capsaicin in inhibiting nlrp3 inflammasome activation

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): Bone marrow cells of C57 mice (purchased from Shanghai Southern Model Bio Co., Ltd., SPF grade 8-week-old male mice) were obtained according to 1×10 6 The density of cells / ml was inoculated into a large dish, and the supernatant containing 20% ​​(v / v) L929 cells (L929 cells were purchased from Shanghai Kebai Company, product number CBP60878, laboratory cultured L929 cells to obtain medium containing secretions) and 10% (v / v) fetal bovine serum (purchased from Shanghai Weike Biotechnology Co., Ltd., product number S1580-500) was cultured in DMEM medium for 5 days, and the DMEM complete medium containing L929 cell supernatant was replaced every 3 days. .

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

Embodiment 2

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

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

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

Embodiment 3

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

[0065] (1) Acquisition of BMDM: the same as step (1) of Example 1.

[0066] (2) In a six-well plate, according to 2 × 10 6 BMDM was seeded overnight at a density of 10 cells / well. 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 with a final concentration of 5 μM was added for 30 min, and the supernatant was removed. NP-40 lysis buffer containing protease inhibitors was added to lyse the cells, and the cells were incubated on ice for 30 min with shaking. Afterwards, a part of the cell lysate was taken as the internal reference for the next immunoprecipitation experiment, and the remaining lysate was centrifuged at 6000 × g for 10 min at 4°C. Take the pellet, wash it three times with PBS buffer, and then...

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Abstract

The invention discloses the application of capsaicin in inhibiting the activation of NLRP3 inflammasome. The present invention finds through research that capsaicin can inhibit the activation of NLRP3 inflammasomes by different agonists such as nigericin, microcrystalline sodium urate and alum, but does not affect the activation of inflammasomes such as NLRC4 and AIM2, and can also inhibit NLRP3 Assembly of the inflammasome, including inhibition of NLRP3 binding to ASC proteins and formation of ASC specks. In addition, the present invention also found that capsaicin can inhibit lipopolysaccharide-induced sepsis. It can be seen that capsaicin can be used as an inhibitor of NLRP3 inflammasome activation and as a potential drug 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] Inflammasome is a cytoplasmic multiprotein complex composed of Nod-like receptor (NLR) family members and PYHIN (pyrin and HIN domain) family members, which can be activated by a variety of pathogen-related molecular patterns or damage-related molecular patterns. The function of the inflammasome is to activate caspase-1 (Caspase-1), which in turn causes the maturation and secretion of the 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 adaptor 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 ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
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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