Application of beta-sitosterol to preparation of medicine for treating or preventing influenza A

A technology of influenza A and influenza A virus, which is applied in the field of medicine, can solve the problems that the effect of β-sitosterol has not been reported, and achieve the effect of meeting the needs of large-scale production, easy quality control, and stable chemical properties

Active Publication Date: 2016-07-20
THE FIRST AFFILIATED HOSPITAL OF GUANGZHOU MEDICAL UNIV (GUANGZHOU RESPIRATORY CENT) +3
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there has been no report on the role of β-sitosterol in inhibiting the inflammatory response mediated by influenza A virus

Method used

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  • Application of beta-sitosterol to preparation of medicine for treating or preventing influenza A
  • Application of beta-sitosterol to preparation of medicine for treating or preventing influenza A
  • Application of beta-sitosterol to preparation of medicine for treating or preventing influenza A

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0022] Example 1 Effect of β-sitosterol on Influenza Virus Induced Host Signaling Pathway

[0023] 1. Materials: cells and viruses

[0024] A549 cells were purchased from ATCC and kept in our laboratory; A / PR8 / 3 / 4(H1N1) were kept in our laboratory.

[0025] 2. Chemical reagents and antibodies

[0026] Trisbase was purchased from Ameresco; sodium chloride (NaCl) was purchased from Guangzhou Chemical Reagent Factory; potassium chloride (KCl) was purchased from Guangzhou Chemical Reagent Factory; potassium dihydrogen phosphate (KH 2 PO 4 ) was purchased from Guangzhou Chemical Reagent Factory; disodium hydrogen phosphate (Na 2 HPO 4 12H 2 O) was purchased from Guangzhou Chemical Reagent Factory; Glycine was purchased from Guangzhou Chemical Reagent Factory; Sodium Dodecyl Sulfonate (SDS) was purchased from American Sigma Company; Tween Tween-20 was purchased from Guangzhou Chemical Reagent Factory; BCA protein concentration determination Kit was purchased from American Ther...

Embodiment 2

[0050] Example 2 Effect of β-sitosterol on the stimulation signal transduction of exogenous interferon (IFN-β)

[0051] A549 cells were seeded into 6-well plates and placed in an incubator. After the cells adhered to the wall after culturing for 12 hours, the original medium was discarded, washed twice with PBS, and then added with influenza virus A / PR8 / 3 / 4 (H1N1) without Serum DMEM / DF12 (1:1) medium adsorbed the cells for 2 hours; then, the cells were washed with PBS to remove unadsorbed virus, and different concentrations of β-sitosterol (200 μg / ml, 400 μg / ml, 600 μg / ml) were added , continue to incubate for 4 hours. After adding 20ng / ml IFN-β to stimulate for 15min, the total protein of the cells was extracted for western blot (the specific process can be carried out by referring to the corresponding steps in Example 1). In this example, rabbit anti-human primary antibodies P-STAT1 and P-STAT3 antibodies were used to incubate the antibodies To detect P-STAT1 and P-STAT3 mo...

Embodiment 3

[0052] Example 3 Effect of β-sitosterol on Influenza Virus Infection Induced Inflammatory Mediators

[0053] A549 cells were planted into 6-well plates, placed in an incubator, and cultured for 12 hours. After the cells adhered to the wall, the original medium was discarded, washed twice with PBS, and then added with influenza virus A / PR8 / 3 / 4 (H1N1) without The DMEM / DF12 (1:1) medium of serum adsorbed the cells for 2 hours; then, the cells were washed with PBS to remove unadsorbed viruses. Using drug treatment mode, drug intervention group added different concentrations of β-sitosterol (150μg / ml, 300μg / ml, 450μg / ml). 300 μg / ml of β-sitosterol was added to the drug group alone, and the A549 cells in the drug group alone were not adsorbed by influenza virus. After continuing to culture for 24 hours, the cell culture supernatant was collected, centrifuged to remove cell debris and filtered to remove virus particles. Then the treated culture medium was added to stimulate another...

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PUM

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Abstract

The invention provides a new use of beta-sitosterol in preparation of a medicine for treating or preventing influenza A, and further provides the medicine for treating or preventing the influenza A. The medicine comprises an active component of the beta-sitosterol. The inventor of the application finds that the beta-sitosterol has a significant inhibitory activity for an inflammatory reaction mediated by influenza A virus. The fluorescence real-time quantitative polymerase chain reaction (PCR) is used for confirming that the beta-sitosterol can obviously inhibit abnormal expressions of A549 inflammatory cytokines infected by an H1N1 virus and show a dose-dependent relationship; the Western blotting is adopted to confirm that the beta-sitosterol can inhibit activation with host inflammation related signaling pathways and show the dose-dependent relationship. Animal experiments show that the beta-sitosterol can significantly inhibit lung injuries and inflammations induced by influenza virus A (H1N1).

Description

technical field [0001] The invention belongs to the field of medicine and relates to the application of a natural product in the preparation of anti-influenza drugs, in particular to the new application of β-sitosterol in the preparation of drugs for treating or preventing influenza A. Background technique [0002] Influenza (abbreviated as flu) is an acute respiratory disease caused by influenza virus, which is highly contagious. Human influenza viruses are divided into three types: A, B, and C. Among them, influenza A is the most threatening to humans, and H1N1 is the most important seasonal influenza A virus. The global annual incidence of influenza is about 5%-10% in adults and 20%-30% in children, and it is easy to cause hospitalization and death in high-risk groups such as infants, the elderly or patients with chronic diseases. Influenza causes about 3 million to 5 million cases of severe disease and about 250,000 to 500,000 deaths every year in the world, seriously t...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K31/575A61P31/16
CPCA61K31/575
Inventor 杨子峰钟南山李菁周倍贤潘锡平关文达李征途李润峰姜志宏
Owner THE FIRST AFFILIATED HOSPITAL OF GUANGZHOU MEDICAL UNIV (GUANGZHOU RESPIRATORY CENT)
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