Figure 1
From: Aspirin inhibits LPS-induced macrophage activation via the NF-κB pathway
ASA pre-treatment inhibited the activation of LPS-induced in mouse peritoneal macrophage cells. (A) The result of flow cytometric analysis showed that >90% of the cells were macrophages after separation. (B) Schematic representation showed the timing of ASA and LPS treatment in our experiments. (C) The results of western blot showed that before the LPS treatment (1 µg/ml, 24 hrs), the addition of ASA pre-treatment at 200 µg/ml decreased the protein expression of total iNOS. Full-length gels are presented in Supplementary Figure 6. (D) The result of PCR showed that 200 µg/ml ASA could significantly decrease the LPS-induced expression of iNOS. We also found ASA below 200 µg/ml concentration could downregulate the expression levels of iNOS induced by LPS. However, 200 µg/ml aspirin downregulated the expression level of iNOS more stablely when compared to the other concentrations. (E) The result of ELISA showed the downregulation of TNF-α after ASA pre-treatment (200 µg/ml). (F) Immunocytochemical staining assays showed the LPS-induced iNOS positive cells ratio signaficantly increased after LPS inducement (77.79% ± 3.22%) compared to the control group (11.26% ± 1.39%), and 200 µg/ml ASA pre-treatment made the LPS-induced iNOS positive cells ratio decrease (12.29% ± 2.36%). Scale bar = 50 µm. All results are representative of at least three independent experiments. Results were expressed as mean ± standard deviation (SD), and statistical significance was shown as N P > 0.05, *P < 0.05 or **P < 0.01.
