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Application of Globulol

A technology of cytokines and compositions, applied in the application field of Globuol, can solve the problems of affecting, reducing the quality of life of patients, and threatening life.

Pending Publication Date: 2021-09-10
GUANGDONG UNIV OF TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

COVID-19 will have a serious impact on the patient's respiratory system, especially the lung tissue, and greatly reduce the quality of life of the patient, which is even life-threatening. Therefore, it is urgent to develop a drug that inhibits acute lung injury caused by cytokines. anti-inflammatory drug

Method used

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  • Application of Globulol
  • Application of Globulol
  • Application of Globulol

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029] Embodiment 1MTT experiment

[0030] MTT assay was used to determine the combined experiment of cytokines γ-IFN, TNF-α and IL-6 and the influence of Globulol on the growth of BEAS-2B cells. The specific implementation plan is as follows:

[0031] 1. Take the BEAS-2B cells in a good growth state, count them with a cell counting board after blowing down, and adjust the final concentration of cells to 5×10 4 pieces / ml. Add 100 μl of cell suspension per well in a 96-well plate at 37 °C and 5% CO 2 After culturing in the incubator for 24 hours, discard the old medium;

[0032] 2. Set up three groups of experiments, respectively (1) set the concentration of γ-IFN, TNF-α and IL-6 to 100ng / ml, 60ng / ml and 10ng / ml respectively, (2) set the concentration of Globulol to 2μM, 5μM, 10μM, 20μM, (3) set cytokine combination (γ-IFN 100ng / ml, TNF-α60ng / ml, IL-6 10ng / ml) in combination with different concentrations of Globulol (2μM, 5μM, 10μM, 20μM). Set up 4 replicate wells for each ...

Embodiment 2

[0034] Embodiment 2 trypan blue experiment

[0035] Use the trypan blue experiment to measure the influence of the combined use of cytokines γ-IFN, TNF-α and IL-6 on the proliferation of BEAS-2B cells. The specific implementation is as follows:

[0036] 1. Take the BEAS-2B cells in good growth state, count them with a cell counting board after blowing down, and adjust the final concentration of cells to 0.2×10 5 pieces / ml. Add 2ml of cell suspension per well in a 35mm cell culture dish and place at 37°C and 5% CO 2 After culturing in the incubator for 24 h, the old medium was discarded.

[0037] 2. Set up two groups of experiments respectively, respectively (1) set the concentrations of γ-IFN, TNF-α and IL-6 as 100ng / ml, 60ng / ml and 10ng / ml respectively, (2) set the combination of cytokines (γ- IFN 100ng / ml, TNF-α 60ng / ml, IL-6 10ng / ml) were used in combination with different concentrations of Globulol (2μM, 5μM, 10μM, 20μM), and 4 replicate wells were set for each concentr...

Embodiment 3

[0039] Embodiment 3 Caspase-1 activity experiment

[0040] Take the BEAS-2B cells in good growth state, count them with a cell counting board after blowing down, and adjust the final concentration of cells to 0.2×10 5 pieces / ml. Add 5ml of cell suspension to a 60mm cell culture dish, place it in an incubator at 37°C and 5% CO2 for 24 hours, and then discard the old medium.

[0041] Set up two groups of experiments respectively, respectively set (1) γ-IFN, TNF-α and IL-6 concentrations are 100ng / ml, 60ng / ml and 10ng / ml respectively (2) set up cytokine combination (γ-IFN 100ng / ml , TNF-α60ng / ml, IL-610ng / ml) combined with different concentrations of Globulol (2μM, 5μM, 10μM, 20μM). After cell culture for 72 hours, trypsinize the adherent cells, centrifuge at 600g at 4°C for 5 minutes to collect the cells, carefully aspirate the supernatant while ensuring that no cells are aspirated as much as possible, and wash once with PBS. After aspirating the supernatant as before, add th...

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Abstract

The invention provides a cell factor composition and an application thereof. The cell factor composition comprises gamma-IFN, TNF-alpha and IL-6. The invention further provides application of Globulol. The Globulol can significantly reduce the pyroptosis and death of pulmonary epithelial cells BEAS-2B induced by the gamma-IFN, the TNF-alpha and the IL-6.

Description

technical field [0001] The present invention relates to a cytokine composition and its application, and also relates to the application of Globulol. Background technique [0002] Novel coronavirus pneumonia (COVID-19) is an acute respiratory infectious disease caused by a new type of coronavirus infection, mainly manifested as fever, dry cough, fatigue and other clinical manifestations. The immune system of severe or critically ill patients with COVID-19 is abnormally activated, and immune cells release a large number of cytokines, causing a severe systemic inflammatory response, triggering a cytokine storm, leading to severe respiratory syndrome, acute lung injury, and even death. Cytokine storm-related cytokines are released when immune cells are activated and interact with target cells. Related cytokines (γ-IFN, TNF-α and IL-6, etc.) are mainly derived from activated immune cells. And it re-acts on the activation of immune cells (including T-cells, monocytes and macropha...

Claims

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Application Information

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IPC IPC(8): A61K38/21A61P11/00A61K38/20A61K38/19
CPCA61K38/217A61K38/191A61K38/204A61P11/00A61K2300/00
Inventor 赵凯郑希李雪彤杨静茹黄泽彬李春莲张蓝月赵肃清郑俊霞何燕黄华容
Owner GUANGDONG UNIV OF TECH
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