Medicinal application of pristimerin as inflammatory cytokine inhibitor
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Embodiment 1
[0021] Example 1 Prissine inhibits the release of TNF-α, IL-8 and IL-6 in vitro
[0022] Cell culture: use RPMI 1640 culture medium containing 10% fetal bovine serum for THP-1 cell line, add penicillin 100U / mL, streptomycin 100U / mL, place at 37°C, 5% CO 2 Cultured in an incubator.
[0023] Determination of cytokine (TNF-α, IL-6 and IL-8) content: THP-1 cells were counted as 1×10 6 / mL inoculated in a 48-well plate, divided into blank control group (DMSO), LPS group (without adding test samples), physesine group (0.15 μM, 0.3 μM and 0.6 μM) and sub-methasone (Dex) group, 3 parallel wells for each sample. After 4 hours of treatment with pristimerin and Dex, except for the blank control group, 1 μg / mL LPS was added to each well for 12 hours, and the supernatant was collected by centrifugation for the determination of TNF-α, IL-6 and IL-8. Calculate the inhibition rate using the following formula:
[0024]
[0025] The results are shown in Tables 1-3.
[0026] Table 1 phys...
Embodiment 2
[0037] Embodiment 2 physesine inhibits active oxygen free radicals
[0038] Cell culture: use RPMI 1640 culture medium containing 10% fetal bovine serum for THP-1 cell line, add penicillin 100U / mL, streptomycin 100U / mL, place at 37°C, 5% CO 2 Cultured in an incubator.
[0039] Determination of ROS (active oxygen molecule) content: THP-1 cells 1×10 6 / mL inoculated in a 24-well plate, and each group was paralleled to 4 wells. The blank control group (DMSO) and the LPS group (without the test sample) were added with the same amount of solvent, and the pristimerin group was added with 0.15 μM, 0.3 μM and 0.6μM prissine, incubated for 2h, except for the blank control group, added 1μg / mL LPS to each well for 12h, then added 5μM DHR 123 (dihydrorhodamine 123) to each well and incubated for 30min, collected cells, and flowed The cytometry was performed and the results are shown in Table 4.
[0040] Table 4 Effect of physesine on ROS generation
[0041]
[0042]
[0043] Rem...
Embodiment 3
[0045] The influence of embodiment 3 physesine on THP-1 cytotoxicity
[0046] Cell culture: use RPMI 1640 culture medium containing 10% fetal bovine serum for THP-1 cell line, add penicillin 100U / mL, streptomycin 100U / mL, place at 37°C, 5% CO 2 Cultured in an incubator.
[0047] Method: 1×10 4 The cells / well were inoculated in 96-well plates, 0.15 μM, 0.3 μM and 0.6 μM physgenin were added to the administration group, and the same amount of solvent was added to the cells in the blank control group, and cultured at 37 ° C. After 12 hours, each well was added 10 μL of MTT (5mg / mL), after co-incubating for 4 hours, the activity of succinate dehydrogenase in living cells was measured, and the OD value was measured at 490nm, and the results were expressed as inhibition rate. The experimental results are shown in Table 5.
[0048] The influence of table 5 physesine on the proliferation of THP-1 cells
[0049]
[0050] The experimental results showed that prissine had no obvio...
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