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. 2011 Feb 18;2:7. doi: 10.3389/fphar.2011.00007

Figure 2.

Figure 2

Effects of hyperforin on arachidonic acid-induced prostanoid formation in human whole blood. (A,B) Heparinized human whole blood was treated with 10 μg ml−1 LPS for 16 h at 37°C and 5% CO2, supplemented with TX synthase inhibitor CV4151 (1 μM), and pre-incubated with Hyp or vehicle (DMSO) for 10 min at 37°C. (A) Then, PGE2 formation was initiated with 20 μM AA, and PGE2 formed within 10 min was separated by RP-HPLC and quantified by ELISA. The 100% value corresponds to PGE2 levels in the range of 18–31 ng ml−1 in the individual experiments, respectively. (B) 6-keto PGF1α was directly determined in the plasma by ELISA. The 100% value corresponds to 6-keto PGF1α levels in the range of 4–7 ng ml−1. Indomethacin (Indo, 50 μM) and celecoxib (Cele, 20 μM) were used as controls. (C) 12-HHT formation in whole blood. Heparinized whole blood was pre-incubated with Hyp or vehicle (DMSO) for 10 min, and AA (100 μM) and Ca2+-ionophore (30 μM) were added to induce 12-HHT product formation. After 10 min at 37°C, 12-HHT was extracted form the plasma by RP-18 solid phase extraction and analyzed by RP-HPLC as described. The 100% value corresponds to 1.5–2.4 μg ml−1 12-HHT. Indomethacin (Indo, 20 μM) and aspirin (ASA, 30 μM) were used as controls. Data are given as mean ± SE, n = 3–5, *p < 0.05, **p < 0.01 or ***p < 0.001 vs. vehicle (0.1% DMSO) control, ANOVA + Tukey HSD post hoc tests.