Effect of BE-IS on cotton pellet-induced granuloma in rats.

<p>With rats under ether anesthesia, sterile cotton pellets weighing 50±1 mg were implanted subcutaneously in both axillae regions of each rat by a single needle incision. Then rats were divided randomly into five groups: Control group (0.5% CMC-Na, p.o), Dexamethasone (DEX) group (2.5 mg/kg, i.p.) and BE-IS groups (100, 200, and 400 mg/kg, p.o). On day 8, granuloma tissue was carefully dissected. The pellets were incubated at 37°C for 24 h and dried at 60°C to constant weight. The increase in dry weight of the pellets was used to measure granuloma formation. Values are expressed as mean ± S.E., n = 10, *<i>P</i><0.05, **<i>P</i><0.01 as compared to the control group.</p

Effect of BE-IS on rat pleurisy induced by carrageenan.

<p>Each value represents as mean ± S.E., n = 10.</p><p>*<i>P</i><0.05,</p><p>**<i>P</i><0.01,</p><p>***<i>P</i><0.001 compared to control group.</p

Effect of BE-IS on the viability of RAW264.7 cells.

<p>Cells were incubated for 24% DMSO, 1 µg/ml LPS, or BE-IS (1.25, 2.5, 5, 10, 20 µg/ml, dissolved in 0.1% DMSO). Cell viability was determined by MTT assay. The optical density was measured at 550 nm on a microplate reader. Values are expressed as mean ± S.E. for three different experiments performed in triplicate.</p

Sighting study in acute oral toxicity assay–fixed dose procedure.

<p>*Clinical signs and conditions associated with pain, suffering, and impending death, are described in detail in a separate OECD Guidance Document <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0095931#pone.0095931-OECDGuidanceDocumentonthe1" target="_blank">[31]</a>.</p

Effect of BE-IS on complete Freund’s adjuvant-induced chronic arthritic rats.

<p>Each value represents as mean ± S.E., n = 10.</p><p>*<i>P</i><0.05,</p><p>**<i>P</i><0.01 compared to control group.</p

Effect of BE-IS on LPS-induced production of NO (A) and PGE₂ (B) in RAW264.7 cells.

<p>Cells were incubated with BE-IS (0, 1.25, 2.5, 5, 10, 20 µg/ml) for 1 h before stimulation with LPS (1 µg/ml) for 24 h. The concentrations of NO and PGE₂ in the cell supernatants were determined by the Griess reaction and ELISA, respectively, according to the manufacturers’ instructions. Values are expressed as mean ± S.E. for three independent experiments. ^#<i>P</i><0.001 as compared to the control group, *<i>P</i><0.05, **<i>P</i><0.01, ***<i>P</i><0.001 as compared to LPS-alone group.</p

Effect of BE-IS on ear edema (A) and MPO activity (B) induced by croton oil in mice.

<p>Mice were divided randomly into five groups: Control group (0.5% CMC-Na), Ibuprofen group (300 mg/kg) and BE-IS groups (150, 300, and 600 mg/kg). Ear edema was induced by topical application of 1% croton oil on the outer and inner surfaces of the right ear of each mouse. The left ear remained untreated and served as a control. Ear edema and MPO activity were measured 4 h after application of croton oil. Values are expressed as mean ± S.E., n = 20, *<i>P</i><0.05, **<i>P</i><0.01 as compared to the control group.</p

Effect of BE-IS on LPS-induced production of TNF-α (A), IL-1β (B) and IL-6 (C) in RAW264.7 cells.

<p>Cells were incubated with BE-IS (0, 1.25, 2.5, 5, 10, 20 µg/ml) for 1 h before stimulation with LPS (1 µg/ml) for 24 h. The concentrations of TNF-α, IL-1β and IL-6 in the cell supernatants were determined by ELISA according to the manufacturers’ instructions. Values are expressed as mean ± S.E. for three independent experiments. ^#<i>P</i><0.001 as compared to the control group, *<i>P</i><0.05, **<i>P</i><0.01, ***<i>P</i><0.001 as compared to LPS-alone group.</p