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A composition for inhibiting macrophage activation and its application in preparation of anti-inflammatory products

A macrophage and composition technology, applied in the field of anti-inflammatory drugs, can solve problems such as cell apoptosis, and achieve the effects of delaying drug resistance, reducing concentration and reducing damage

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

AI Technical Summary

Problems solved by technology

In addition, undegraded dysfunctional mitochondria generate more ROS and are more likely to release cytochrome c, leading to apoptosis

Method used

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  • A composition for inhibiting macrophage activation and its application in preparation of anti-inflammatory products
  • A composition for inhibiting macrophage activation and its application in preparation of anti-inflammatory products
  • A composition for inhibiting macrophage activation and its application in preparation of anti-inflammatory products

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033] Different concentrations of cinnamide and inhibitor MDIVI-1 on Raw264.7 cells were measured by MTT experiments, as follows:

[0034] Raw264.7 cells were blown down from the culture bottle with a pipette, and cell suspension was made using 10% FBS DMEM medium to inoculate concentration 5 × 10 4 The concentration of / ml is inoculated on a 96-well plate (100 μL / well), placed in 5% CO 2 It was cultured for 24 hours in a 37 ° C incubator. The cinnamide and MdiVi-1 concentration were set to 80 μm, 40 μm, 20 μm, 10 μm, 5 μM, and four additional holes were disposed for each concentration. 8% CO 2 At 37 ° C for 24 h, after suction of the supernatant, 100 μl of concentration of 0.5 mg / ml of MTT solution (diluted with a 5 mg / ml MTT solution in the base DMEM was obtained); inserted into the incubator, continued to cultivate for 3 hours. Be careful to suck the supernatant in the hole. 100 μl of DMSO was added to each well, and the incubator was allowed to stand for ten minutes; t...

Embodiment 2

[0037] Lipopolysaccharide (LPS) is a component of the outer wall of Gram-negative bacterial cells, which has strong inflammation, but there is no direct toxic effect on macrophages. It has been widely used in a variety of inflammatory responses and oxidative stypes, including Raw264.7 cells in vitro model, so the following tests were tested using LPS modeling. Based on LPS (lipid polysaccharide) induction, cinnamarne (CMA) 20 μm, 10 μm and MDIVI-1 30 μm, 20 μm, and 10 μm performed separate and two-two combination drugs to produce NO content in Raw264.7. With the Griess experiment, the specific embodiments are as follows:

[0038] Raw264.7 cells were blown down from the culture flask using a pipette, and cell suspensions were made using 10% FBS DMEM medium to inoculate concentration 1 × 10 6 The concentration of / ml is inoculated on a 24-well plate (400 μL / well), placed in 5% CO 2 , Cultured in a 37 ° C incubator. After 24 hours, the medium, the blank group was added to 400 μl o...

Embodiment 3

[0042] Cinnamonal aldehydrates with Western blot experiments promote primary and autophagy, and specific embodiments are as follows:

[0043] 1. Cell treated before the protein: EtOAc. After 24 hours, the supernatant was discarded, and 2 ml of the fully medium containing cinnamarcal aldehyde (20 μm) was added. After 2 hours, LPS was added to give a final concentration of 1 μg / ml while setting the blank group and the LPS model group. After 24 hours of continuing, cell extraction can be performed.

[0044] 2. Cell total protein extract method: Raw264.7 cells in petri dishes were gently washed three times with PBS, transferred to 1.5 ml EP tube, centrifuged at low speed, and discarded PBS. The EP tube was placed on ice, and 100 μl of cell lysate was added, mixed with lysis of 10 minutes. The resulting supernatant is a cell total protein solution at a high speed and low temperature (4 ° C / 12000 rpm).

[0045] 3. Determination of protein concentration: Formulated protein standard s...

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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 its application in preparing anti-inflammatory products. The composition is composed of cinnamaldehyde and Drp1 inhibitor. The molar ratio of the cinnamaldehyde to the Drp1 inhibitor is (0.3-2.3): (0.5-3.5). The composition has good anti-inflammation effect, utilizes the synergistic effect of the cinnamaldehyde and the Drp1 inhibitor, can reduce the concentration of the single drug, thereby reducing the toxic and side effects of the single drug and reducing the damage to the human body. The results of the present invention show that cinnamaldehyde can promote mitophagy, but has no inhibitory effect on mitochondrial fission, and the Drp1 inhibitor can inhibit mitochondrial fission. Therefore, the combination of cinnamaldehyde and Drp1 inhibitors can inhibit the inflammatory response by acting on different targets, and it is expected to become a potential therapy for the treatment of inflammation-related diseases.

Description

Technical field [0001] The invention belongs to the technical field of anti-inflammatory drugs, and more particularly to a composition inhibiting macrophage activation and its application in preparing anti-inflammatory products. Background technique [0002] Macrophages are an important involvement and regulators of the body inflammatory response as an initial immune cell. Macrophages can remove invasive pathogens, trigger inflammatory signals and swallow dead cells. More and more evidence suggests that macrophages are necessary to grow and maintain metabolic stability based on different tissues of macrophage a phagocytosis and cytokine signal adjustment. Macrophages play a double role during injury and pathogens. In many diseases, such as cancer, inflammatory diseases, fibrosis, etc., macrophages are considered greater role in disease progression when inflammatory macrophages cannot be inhibited. Under different environmental conditions, it can differentiate into classical activ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
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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