Application of pterostilbene in anti-influenza virus drugs
A type of influenza A virus, influenza virus technology, applied in the direction of antiviral agents, ether/acetal active ingredients, etc., can solve problems such as universal drug resistance
- Summary
- Abstract
- Description
- Claims
- Application Information
AI Technical Summary
Problems solved by technology
Method used
Image
Examples
Embodiment 1
[0036] Pterostilbene for respiratory epithelial cells and toxicity assessment:
[0037] A549 cells were plated at 1 × 10 4 Inoculated in a 96-well plate / well at 37°C, 5% CO 2 Incubate overnight in incubator;
[0038] The supernatant was discarded from the cells, and the doubling-diluted pterostilbene was added to the lung epithelial cells (A549);
[0039] The cytotoxicity of pterostilbene on airway epithelial cells was detected by MTT method: After 48 hours, the supernatant was discarded, and the medium containing 0.5 mg / ml MTT was added to each well, and after incubation at 37 °C for 4 hours, the microplate reader was used to detect the OD570 nm. absorbance.
[0040] like figure 2 As shown, the experimental results showed that pterostilbene was less toxic to A549 cells, and the cell viability was >75% at the maximum concentration of 200 μM.
Embodiment 2
[0042] Evaluation of pterostilbene for respiratory epithelial cells and the ability to inhibit virus infection:
[0043] 1) Divide A549 cells into 3 x 10 5 Inoculated in a 6-well plate / well at 37°C, 5% CO 2 Incubate overnight in incubator;
[0044] 2) Discard the supernatant from the cells, dilute the virus with serum-free RPMI1640 medium, inoculate A549 cells with a virus titer of MOI=1, and adsorb in a 37°C, 5% CO2 incubator for 1 hour;
[0045] 3) After virus infection, discard the virus supernatant, add pterostilbene 40 times diluted in serum-free RPMI16 to the cells, at 37°C, 5% CO 2 Continue to cultivate in the incubator;
[0046] 4) After 24 hours, a) collect cell proteins, and use western blot to detect the expressions of influenza virus-related proteins NP, M2 and PB2. The results are as follows image 3 Compared with the virus control group, the expression levels of influenza virus proteins NP, PB2 and M2 were lower than those in the control group after adding pt...
Embodiment 3
[0051] Pterostilbene is used for respiratory epithelial cells and detection of NP expression:
[0052] 1) A549 cells at 2 x 10 4 Cells / well were seeded in confocal Petri dishes under 5% CO. 2 , cultured in a cell incubator at 37°C for 24h;
[0053] 2) Discard the culture supernatant, wash twice with PBS, add 200 μL of virus solution (MOI=1), in 5% CO 2 , cells were adsorbed in a cell incubator at 37°C for 1 h;
[0054] 3) Discard the virus solution, add the compound of the specified concentration to the petri dish, set up cell control wells and virus control wells, and continue to culture for 24 hours;
[0055] 4) Discard the supernatant and wash 3 times with pre-cooled PBS;
[0056] 5) Fixation and blocking of cells: add 200 μL of 4% paraformaldehyde to each well for 20 minutes, wash 3 times with PBS, and block with 200 μL of 3% bovine serum albumin for 1 hour at room temperature;
[0057] 6) Incubation with primary antibody: discard the blocked bovine serum albumin solu...
PUM
Login to View More Abstract
Description
Claims
Application Information
Login to View More