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. 2016 Apr 30;49(4):232–237. doi: 10.5483/BMBRep.2016.49.4.002

Fig. 4. RME inhibits LPS-induced BLT2 ligand production in Raw264.7 cells. (A) Raw264.7 cells were incubated with RME (47.5 μg/ml) or OXY (7.3 μg/ml) for 30 min, and further incubated for 12 h in the presence or absence of LPS (100 ng/ml). The quantities of LTB4 (left) and 12(S)-HETE (right) in the culture supernatants were measured by ELISA. (B) Raw264.7 cells were incubated for 30 min with RME or OXY, followed by 3 h incubation in the absence or presence of LPS. Next, the cells were assayed for cPLA2 activity by cPLA2 assay kit. (C) Raw264.7 cells were incubated for 30 min with RME or OXY, and then incubated for 3 h in the absence or presence of LPS. The cells were then evaluated for p-cPLA2, cPLA2, 5-LO and 12-LO protein levels by western blot assay (densitometry data also shown). (D) Cells were treated as detailed in (C), and the cells were evaluated for p-p38 and p38 protein levels by Western blot (densitometry data also shown). (E) Raw264.7 cells were initially incubated for 30 min with AACOCF3 (20 μM), AA861 (10 μM) or baicalein (20 μM), followed by 4 h incubation in the presence or absence of LPS. Subsequently, total RNA was isolated and subjected to RT-PCR analysis. (F) Raw264.7 cells were initially incubated for 30 min with AACOCF3, AA861 or baicalein, and further incubated for 12 h in the presence or absence of LPS; the IL-6 released into the culture medium was then measured. (G) The schematic representation of the mechanism of action by which RME exerts its inhibitory effects on LPS-induced inflammatory responses in Raw264.7 cells. All of the quantitative data are presented as the mean ± the SD, of three independent experiments. *P < 0.05, **P < 0.01, ***P < 0.005.

Fig. 4.