Composition for inhibiting macrophage activation and application thereof in preparation of anti-inflammatory product

A technology of macrophages and compositions, applied in the field of anti-inflammatory drugs, can solve problems such as cell apoptosis, and achieve the effects of delaying drug resistance, reducing NO production, and good anti-inflammatory effects

Active Publication Date: 2021-10-12
GUANGDONG UNIV OF TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

In addition, undegraded dysfunctional mitochondria generate more RO

Method used

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  • Composition for inhibiting macrophage activation and application thereof in preparation of anti-inflammatory product
  • Composition for inhibiting macrophage activation and application thereof in preparation of anti-inflammatory product
  • Composition for inhibiting macrophage activation and application thereof in preparation of anti-inflammatory product

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033] MTT assay was used to measure the growth effects of different concentrations of cinnamaldehyde and inhibitor Mdivi-1 on RAW264.7 cells. The specific implementation scheme is as follows:

[0034] Use a pipette gun to blow off the RAW264.7 cells from the culture flask, use 10% FBS in DMEM medium to make a cell suspension, and inoculate at a concentration of 5×10 4 Inoculated in 96-well plates (100 μL / well) at the concentration of cells / mL, placed in 5% CO 2 , and cultured in a 37°C incubator for 24 hours. The concentrations of cinnamaldehyde and Mdivi-1 were set to 80 μM, 40 μM, 20 μM, 10 μM, and 5 μM, and 4 replicate wells were set for each concentration. 8% CO 2 , cultured at 37°C for 24 hours, sucked off the supernatant, and added 100 μL of MTT solution with a concentration of 0.5 mg / mL (diluted 10 times with 5 mg / mL MTT solution in basic DMEM); put it into the incubator and continue to cultivate for 3 hours, Carefully aspirate the supernatant from the wells. Add 1...

Embodiment 2

[0037] Lipopolysaccharide (LPS), a component of the outer cell wall of Gram-negative bacteria, has a strong inflammatory effect, but has no direct toxic effect on macrophages. It has been widely used in the establishment of various inflammatory response and oxidative stress models, including the RAW264.7 cell in vitro inflammation model, so the following experiments were conducted using LPS modeling. Based on LPS (lipopolysaccharide) induction, the effects of cinnamaldehyde (CMA) 20 μM, 10 μM and Mdivi-1 30 μM, 20 μM, 10 μM alone or in combination on the NO content produced by RAW264.7. Through the Griess experiment, the specific implementation plan is as follows:

[0038] Use a pipette gun to blow off the RAW264.7 cells from the culture flask, use 10% FBS DMEM medium to make a cell suspension, and inoculate at a concentration of 1×10 6 The concentration of cells / mL was inoculated in 24-well plate (400 μL / well), placed in 5% CO 2 , cultured in a 37°C incubator. After 24 hou...

Embodiment 3

[0042] The cinnamaldehyde verified by Western Blot experiment promotes mitophagy, and the specific implementation scheme is as follows:

[0043] 1. Cell treatment before protein extraction: RAW264.7 cells were planted in 35mm culture dishes at 1 million cells per dish, and 8ml of complete medium was added. After 24 hours, the supernatant was discarded, and 2 ml of complete medium containing cinnamaldehyde (20 μM) was added to each dish. After culturing for 2 hours, LPS was added to make the final concentration 1 μg / ml, and a blank group and an LPS model group were set at the same time. After continuing to culture for 24 hours, cell extraction can be performed.

[0044] 2. Extraction method of total cell protein: RAW264.7 cells in the culture dish were gently washed three times with PBS, blown down, transferred to 1.5ml EP tube, centrifuged at low speed, discarded PBS. Place the EP tubes on ice, add 100 μl cell lysate to each tube, mix and lyse for 10 minutes. Afterwards, ce...

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Abstract

The invention belongs to the technical field of anti-inflammatory drugs, and discloses a composition for inhibiting macrophage activation and application thereof in preparation of an anti-inflammatory product. The composition is prepared from cinnamyl aldehyde and a Drp1 inhibitor, wherein the molar ratio of the cinnamyl aldehyde to the Drp1 inhibitor is (0.3-2.3): (0.5-3.5). The composition has a good anti-inflammatory effect, and the concentration of a single drug can be reduced by utilizing the synergistic effect of the cinnamyl aldehyde and the Drp1 inhibitor, so that the toxic and side effects of the single drug are reduced, and the harm to a human body is reduced. The result shows that the cinnamyl aldehyde can promote mitochondrial autophagy, but has no inhibitory effect on mitochondrial fission, and the Drp1 inhibitor can inhibit the mitochondrial fission. Therefore, the cinnamyl aldehyde and the Drp1 inhibitor can be combined to inhibit inflammatory reactions by acting on different targets, and are expected to become potential therapies for treating inflammation-related diseases.

Description

technical field [0001] The invention belongs to the technical field of anti-inflammatory drugs, and more specifically relates to a composition for inhibiting the activation of macrophages and its application in the preparation of anti-inflammatory products. Background technique [0002] As an initial immune cell, macrophage is an important participant and regulator of the body's inflammatory response. Macrophages clear invading pathogens, trigger inflammatory signals and engulf dead cells. Accumulating evidence indicates that macrophages are required for the growth and maintenance of metabolic homeostasis in diverse tissues based on the regulation of macrophage phagocytosis and cytokine signaling. Macrophages play a dual role during injury and pathogen invasion. In many diseases, such as cancer, inflammation-related diseases, fibrosis, etc., when inflammatory macrophages cannot be suppressed, macrophages are considered to play a greater role in disease progression. Under ...

Claims

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Application Information

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IPC IPC(8): A61K31/11A61K45/06A61P29/00A61K31/517A61K31/7048A61K31/166
CPCA61K31/11A61K45/06A61P29/00A61K31/517A61K31/7048A61K31/166
Inventor 许泳瑜卢宇靖黄泽彬钟家本张创
Owner GUANGDONG UNIV OF TECH
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