Application of digoxin in preparation of medicine for treating and/or preventing macrophage M1 type polarization related diseases

A technology of macrophages and digoxin, which is applied in the field of medicine, can solve the problems of no research reports on the treatment of OA with DIG, and achieve the effect of delaying cartilage damage and synovitis

Active Publication Date: 2022-02-01
HAINAN MEDICAL COLLEGE
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

So far, there is no research report on the treatment of OA with DIG

Method used

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  • Application of digoxin in preparation of medicine for treating and/or preventing macrophage M1 type polarization related diseases
  • Application of digoxin in preparation of medicine for treating and/or preventing macrophage M1 type polarization related diseases
  • Application of digoxin in preparation of medicine for treating and/or preventing macrophage M1 type polarization related diseases

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0024] Example 1: DIG inhibits the M1 polarization of synovial macrophages and delays articular cartilage damage

[0025] 1. MTT assay was used to detect the effect of DIG on the activity of mouse macrophage RAW 264.7, and the concentration used to inhibit cell inflammation was screened.

[0026] The specific operation is: spread 96-well plates with 3× concentration of macrophages, add different concentrations of DIG drugs (20 μM, 40 μM, 60 μM, 80 μM) respectively, add 10 μl of MTT solution to each well after 24 hours of treatment, and incubate at 37°C for 2- After 4 hours, 200 [mu]l DMSO was added to each well. Shake at low speed on a shaker at room temperature for 10 minutes to fully dissolve the crystals, and measure the absorbance of each well at OD 490nm in an enzyme-linked immunosorbent assay instrument.

[0027] Survival cells % = (OD experiment / OD control) x 100%

[0028] MTT assay was used to detect the effect of DIG on the activity of mouse macrophage RAW 264.7, an...

Embodiment 2

[0038] Example 2: Isolation, purification and functional identification of exosomes produced by M1 macrophages before and after DIG treatment

[0039]1. Cultivate RAW 264.7 cells, respectively set up blank group, LPS treatment group (1 μg / ml), LPS (1 μg / ml) + DIG (60 μM) treatment group, and ultracentrifugation to separate the exosomes (Exos) in the supernatant of cultured cells ). The exosome particle size was detected by NTA, the shape was detected by transmission electron microscopy, and the exosome Marker (CD9, TSG 101) was detected by Western blot.

[0040] The shape of exosomes produced by M1 macrophages before and after DIG treatment was detected by transmission electron microscopy ( image 3 A), the particle size of exosomes detected by NTA ( image 3 B), Western blot detection of exosome Marker ( image 3 C).

[0041] 2. Isolate primary mouse chondrocytes, stain and mark exosomes derived from macrophages with PKH67, and co-incubate with primary chondrocytes to obs...

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Abstract

The invention discloses application of digoxin in preparation of a medicine for treating and / or preventing macrophage M1 type polarization related diseases. Experiments prove that digoxin promotes macrophages to generate specific exosomes while inhibiting M1 type polarization of synovial macrophages, and osteoarthritis cartilage injury and synovitis can be effectively delayed. The invention provides a treatment mechanism of digoxin for treating osteoarthritis, and provides a new thought for clinical treatment of osteoarthritis.

Description

technical field [0001] The invention belongs to the technical field of medicine, and specifically relates to the use of digoxin in the preparation of medicines for treating and / or preventing macrophage M1 type polarization-related diseases. Background technique [0002] Osteoarthritis (OA) is a joint disease with high incidence in people over 50 years old, often involving knees, hips and other joints. It is estimated that by 2030, its prevalence rate will be as high as 40%, and OA will become the largest cause of disability. Cartilage damage is the main lesion of OA, and the key cause is increased M1 polarization of synovial macrophages. In healthy synovial tissue, the main function of macrophages present in the synovial lining layer is to maintain joint homeostasis. Under the pathological state of OA, macrophages are activated into M1 type macrophages through the classical pathway, namely pro-inflammatory macrophages. Previous research results published by our research te...

Claims

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Application Information

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IPC IPC(8): A61K31/7048A61P29/00A61P19/02A61P19/04
CPCA61K31/7048A61P29/00A61P19/02A61P19/04
Inventor 王华刘让如贾浩
Owner HAINAN MEDICAL COLLEGE
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