Application of apatinib in preparation of medicine for preventing and treating NLRP3 inflammasome related diseases
A technology of inflammasome and apatinib, applied in the field of medicine, can solve problems such as deficiency, and achieve the effect of inhibiting maturation and secretion, and inhibiting the activation of NLRP3 inflammasome
- Summary
- Abstract
- Description
- Claims
- Application Information
AI Technical Summary
Problems solved by technology
Method used
Image
Examples
Embodiment 1
[0050] Example 1 Enzyme-linked immunosorbent assay Apatinib inhibits the release of IL-1β activated by NLRP3 inflammasome
[0051] 1. On the first day, divide J774A.1 cells into 96-well plate, each well 5×10 5 cells;
[0052] 2. Seed the plate overnight, discard the supernatant, add 100 μL of DMEM medium containing 10% serum containing bacterial lipopolysaccharide (LPS) (1 μg / ml) to each well, and then add different concentrations of (0 μM, 0.25 μM, 0.5 μM, 1 μM, 2.5 μM, 5 μM, 10 μM, 20 μM) apatinib treatment for 1 hour, and then add nigericin (Nigericin, 10 μM) for 1 hour;
[0053] 3. After the treatment in step 2, collect the cell supernatant, and use the Mouse IL-1β ELISA kit to measure the IL-1β content. The operation steps are carried out according to the kit instructions, and the inhibition percentage for inhibiting the release of IL-1β is calculated. See the results in figure 1 and figure 2 .
[0054] figure 1 and figure 2 The results showed that under the joint...
Embodiment 2
[0055] Example 2 Determination of Apatinib Inhibition of NLRP3 Inflammasome Activation by Western Blot
[0056] (1) On the first day, J774A.1 cells were inoculated into cell culture dishes with a diameter of 35mm, and each dish contained 5×10 6 cells;
[0057] (2) After 12 hours of cell growth and proliferation in the cell culture dish, the supernatant was discarded, and 1 ml of DMEM medium containing 10% serum containing bacterial lipopolysaccharide (1ug / ml) was added to each dish for 5 hours, and then different concentrations (0μM , 0.25 μM, 0.5 μM) of apatinib for 1 hour, and then add nigericin (Nigericin, 10uM) for 1 hour;
[0058] (3) After the treatment in step (2), collect the cell supernatant and cell lysate, extract the supernatant protein and cell lysate protein from the supernatant and lysate according to the conventional method, and use anti-IL-1β antibody, anti-Caspase-1 antibody , anti-NLRP3 antibody and anti-ASC antibody for Western blotting analysis, see the ...
PUM
Login to View More Abstract
Description
Claims
Application Information
Login to View More