Application of mycoepoxydiene nicotinic acids derivatives in preparation process of pro-inflammatory mediator inhibitors
A technology of epoxy diene and derivatives, applied in the direction of anti-inflammatory agent, drug combination, non-central analgesic, etc., can solve the problem of unreported anti-inflammatory physiological activity and the like
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Embodiment 1
[0022] Embodiment 1 4NDM cytotoxicity test
[0023] The cytotoxicity of deacetylated fungi ethylene oxide was detected by MTT method, and the specific steps were as follows:
[0024] RAW264.7 cells (4×10 5 cells / well) were inserted into a 96-well culture plate and cultured overnight, and bacterial lipopolysaccharide (lipopolysaccharide, LPS) (100 ng / ml) and different concentrations of 4NDM (0.1, 2.5, 5, 7.5, 10, 12, 15 μmol / L, Use DMSO as a blank control) to act together for 24h or different concentrations of 4NDM (0.1, 2.5, 5, 7.5, 10, 12, 15μmol / L, use DMSO as a blank control) to act alone for 24h, and then add MTT to a final concentration of 0.5mg / L ml for 3 hours, add solution (10% SDS, 0.01mol / L HCL) at 100 μl / well overnight, then measure OD 595 .
[0025] OD 595 Can reflect the growth of cells, so by comparing OD 595 Value to judge whether 4NDM has cytotoxicity.
[0026] The results of the MTT assay showed that (see figure 1 , 2 ): 4NDM probably has no cytotoxici...
Embodiment 2
[0027] Example 2 Application of 4NDM in inhibiting the inflammatory response of RAW264.7 cells induced by bacterial lipopolysaccharide
[0028] By detecting the changes in the concentration of pro-inflammatory factors in cells, it can be determined whether 4NDM can inhibit the inflammatory response of RAW264.7 cells. The specific test method is as follows:
[0029] 1) Set the 4NDM concentration gradient. RAW264.7 cells (4×10 5 Cells / mL) were inserted into a 12-well culture plate, and the medium was changed once after 24 h. After 1-2 h, LPS was added to a final concentration of 100 ng / ml to stimulate for 6 h. At the same time, different concentrations of 4NDM (0, 0.1, 1, 5, 10 μmol / L, DMSO was used as a blank control), the effect of 4NDM on the expression of inflammatory factors before LPS stimulation was observed, and the cell culture supernatant was extracted for enzyme-linked immunosorbent assay. Through ELISA detection kit (purchased from eBioscience Company, Mouse TNF-α ...
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