Figure 1.

(A) HPLC characterization of the five fractions. A total of 10 μL each sample (1 mg/mL) was injected and analyzed using a Dionex UltiMate^TM 3000 HPLC system with photodiode array detection (PAD) at 259 nm. A Symmetrix ODS-RC18 (25 × 4.6 mm, 5 mm) HPLC column protected with a Phenomenex security guard column (C18, 4 × 3.0 mm) operated at 30 °C was used, and the flow rate was maintained at 1 mL/min. The elution solvents were acetonitrile (a) and 0.1% acetic acid (b). Samples were eluted according to the following gradient: 0–35 min 30% a isocratic, 35–45 min 30% to 40% a, 45–55 min 40% a isocratic, and finally washing and recondition of the column. (B) Relative expression of IL-1β mRNA after treatment with F40. RAW264.7 cells were pretreated with different concentration of F40 for 1 h and then treated with 200 ng/mL LPS for 6 h. Total RNA was extracted and genes expression level were analyzed by RT-PCR in triplicate. The expression level of each gene was normalized to glyceraldehyde-3-phosphate dehydrogenase (GAPDH) mRNA. Data are expressed as mean ± SD (n = 6). ** p < 0.01 vs. LPS treatment group.