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Hyperforin, an Anti-Inflammatory Constituent from St. John's Wort, Inhibits Microsomal Prostaglandin E(2) Synthase-1 and Suppresses Prostaglandin E(2) Formation in vivo.

Koeberle A, Rossi A, Bauer J, Dehm F, Verotta L, Northoff H, Sautebin L, Werz O - Front Pharmacol (2011)

Bottom Line: The acylphloroglucinol hyperforin (Hyp) from St. John's wort possesses anti-inflammatory and anti-carcinogenic properties which were ascribed among others to the inhibition of 5-lipoxygenase.Intraperitoneal (i.p.) administration of Hyp (4 mg kg(-1)) to rats impaired exudate volume and leukocyte numbers in carrageenan-induced pleurisy associated with reduced PGE(2) levels, and Hyp (given i.p.) inhibited carrageenan-induced mouse paw edema formation (ED(50) = 1 mg kg(-1)) being superior over indomethacin (ED(50) = 5 mg kg(-1)).We conclude that the suppression of PGE(2) biosynthesis in vitro and in vivo by acting on mPGES-1 critically contributes to the anti-inflammatory efficiency of Hyp.

View Article: PubMed Central - PubMed

Affiliation: Department for Pharmaceutical Analytics, Pharmaceutical Institute, University of Tübingen Tübingen, Germany.

ABSTRACT
The acylphloroglucinol hyperforin (Hyp) from St. John's wort possesses anti-inflammatory and anti-carcinogenic properties which were ascribed among others to the inhibition of 5-lipoxygenase. Here, we investigated whether Hyp also interferes with prostanoid generation in biological systems, particularly with key enzymes participating in prostaglandin (PG)E(2) biosynthesis, i.e., cyclooxygenases (COX)-1/2 and microsomal PGE(2) synthase (mPGES)-1 which play key roles in inflammation and tumorigenesis. Similar to the mPGES-1 inhibitors MK-886 and MD-52, Hyp significantly suppressed PGE(2) formation in whole blood assays starting at 0.03-1 μM, whereas the concomitant generation of COX-derived 12(S)-hydroxy-5-cis-8,10-trans-heptadecatrienoic acid, thromboxane B(2), and 6-keto PGF(1α) was not significantly suppressed up to 30 μM. In cell-free assays, Hyp efficiently blocked the conversion of PGH(2) to PGE(2) mediated by mPGES-1 (IC(50) = 1 μM), and isolated COX enzymes were not (COX-2) or hardly (COX-1) suppressed. Intraperitoneal (i.p.) administration of Hyp (4 mg kg(-1)) to rats impaired exudate volume and leukocyte numbers in carrageenan-induced pleurisy associated with reduced PGE(2) levels, and Hyp (given i.p.) inhibited carrageenan-induced mouse paw edema formation (ED(50) = 1 mg kg(-1)) being superior over indomethacin (ED(50) = 5 mg kg(-1)). We conclude that the suppression of PGE(2) biosynthesis in vitro and in vivo by acting on mPGES-1 critically contributes to the anti-inflammatory efficiency of Hyp.

No MeSH data available.


Related in: MedlinePlus

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.
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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.

Mentions: Hyperforin may suppress PGE2 formation by interference with LPS-signaling, mPGES-1 expression or liberation of AA as substrate for COX enzymes. Thus, heparinized human whole blood was first incubated with LPS for 16 h to induce the expression of COX-2 and mPGES-1 (in contrast to constitutively expressed COX-1 and cPGES). Then, the blood was pre-incubated with Hyp for 10 min, and prostanoid formation was initiated by addition of 20 μM AA. After 10 min at 37°C, generation of PGE2 and 6-keto PGF1α was determined in plasma as described above. PGE2 formation was potently inhibited by Hyp in a concentration-dependent manner with an IC50 of 0.25 μM reaching significance at 30 nM (Figure 2A), whereas formation of 6-keto PGF1α was not significantly affected up to 30 μM (Figure 2B), again indicating selective suppression of PGE2 biosynthesis by Hyp. Note that Hyp (3–30 μM) was more efficient in suppressing PGE2 synthesis after long-term pre-stimulation with LPS (induction of COX-2 and mPGES-1, Figure 2A) than during short term stimulation (Figure 1B) supporting that Hyp interferes with the inducible COX-2/mPGES-1 pathway.


Hyperforin, an Anti-Inflammatory Constituent from St. John's Wort, Inhibits Microsomal Prostaglandin E(2) Synthase-1 and Suppresses Prostaglandin E(2) Formation in vivo.

Koeberle A, Rossi A, Bauer J, Dehm F, Verotta L, Northoff H, Sautebin L, Werz O - Front Pharmacol (2011)

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.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC3108608&req=5

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.
Mentions: Hyperforin may suppress PGE2 formation by interference with LPS-signaling, mPGES-1 expression or liberation of AA as substrate for COX enzymes. Thus, heparinized human whole blood was first incubated with LPS for 16 h to induce the expression of COX-2 and mPGES-1 (in contrast to constitutively expressed COX-1 and cPGES). Then, the blood was pre-incubated with Hyp for 10 min, and prostanoid formation was initiated by addition of 20 μM AA. After 10 min at 37°C, generation of PGE2 and 6-keto PGF1α was determined in plasma as described above. PGE2 formation was potently inhibited by Hyp in a concentration-dependent manner with an IC50 of 0.25 μM reaching significance at 30 nM (Figure 2A), whereas formation of 6-keto PGF1α was not significantly affected up to 30 μM (Figure 2B), again indicating selective suppression of PGE2 biosynthesis by Hyp. Note that Hyp (3–30 μM) was more efficient in suppressing PGE2 synthesis after long-term pre-stimulation with LPS (induction of COX-2 and mPGES-1, Figure 2A) than during short term stimulation (Figure 1B) supporting that Hyp interferes with the inducible COX-2/mPGES-1 pathway.

Bottom Line: The acylphloroglucinol hyperforin (Hyp) from St. John's wort possesses anti-inflammatory and anti-carcinogenic properties which were ascribed among others to the inhibition of 5-lipoxygenase.Intraperitoneal (i.p.) administration of Hyp (4 mg kg(-1)) to rats impaired exudate volume and leukocyte numbers in carrageenan-induced pleurisy associated with reduced PGE(2) levels, and Hyp (given i.p.) inhibited carrageenan-induced mouse paw edema formation (ED(50) = 1 mg kg(-1)) being superior over indomethacin (ED(50) = 5 mg kg(-1)).We conclude that the suppression of PGE(2) biosynthesis in vitro and in vivo by acting on mPGES-1 critically contributes to the anti-inflammatory efficiency of Hyp.

View Article: PubMed Central - PubMed

Affiliation: Department for Pharmaceutical Analytics, Pharmaceutical Institute, University of Tübingen Tübingen, Germany.

ABSTRACT
The acylphloroglucinol hyperforin (Hyp) from St. John's wort possesses anti-inflammatory and anti-carcinogenic properties which were ascribed among others to the inhibition of 5-lipoxygenase. Here, we investigated whether Hyp also interferes with prostanoid generation in biological systems, particularly with key enzymes participating in prostaglandin (PG)E(2) biosynthesis, i.e., cyclooxygenases (COX)-1/2 and microsomal PGE(2) synthase (mPGES)-1 which play key roles in inflammation and tumorigenesis. Similar to the mPGES-1 inhibitors MK-886 and MD-52, Hyp significantly suppressed PGE(2) formation in whole blood assays starting at 0.03-1 μM, whereas the concomitant generation of COX-derived 12(S)-hydroxy-5-cis-8,10-trans-heptadecatrienoic acid, thromboxane B(2), and 6-keto PGF(1α) was not significantly suppressed up to 30 μM. In cell-free assays, Hyp efficiently blocked the conversion of PGH(2) to PGE(2) mediated by mPGES-1 (IC(50) = 1 μM), and isolated COX enzymes were not (COX-2) or hardly (COX-1) suppressed. Intraperitoneal (i.p.) administration of Hyp (4 mg kg(-1)) to rats impaired exudate volume and leukocyte numbers in carrageenan-induced pleurisy associated with reduced PGE(2) levels, and Hyp (given i.p.) inhibited carrageenan-induced mouse paw edema formation (ED(50) = 1 mg kg(-1)) being superior over indomethacin (ED(50) = 5 mg kg(-1)). We conclude that the suppression of PGE(2) biosynthesis in vitro and in vivo by acting on mPGES-1 critically contributes to the anti-inflammatory efficiency of Hyp.

No MeSH data available.


Related in: MedlinePlus