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Consequences of altered eicosanoid patterns for nociceptive processing in mPGES-1-deficient mice.

Brenneis C, Coste O, Schmidt R, Angioni C, Popp L, Nusing RM, Becker W, Scholich K, Geisslinger G - J. Cell. Mol. Med. (2008)

Bottom Line: Cyclooxygenase-2 (COX-2)-dependent prostaglandin (PG) E(2) synthesis in the spinal cord plays a major role in the development of inflammatory hyperalgesia and allodynia.Microsomal PGE(2) synthase-1 (mPGES-1) isomerizes COX-2-derived PGH(2) to PGE(2).Here, we evaluated the effect of mPGES-1-deficiency on the nociceptive behavior in various models of nociception that depend on PGE(2) synthesis.

View Article: PubMed Central - PubMed

Affiliation: Pharmazentrum Frankfurt, ZAFES, Institut für Klinische Pharmakologie, Klinikum der Johann Wolfgang Goethe-Universität Theodor-Stern-Kai 7, Frankfurt, Germany.

ABSTRACT
Cyclooxygenase-2 (COX-2)-dependent prostaglandin (PG) E(2) synthesis in the spinal cord plays a major role in the development of inflammatory hyperalgesia and allodynia. Microsomal PGE(2) synthase-1 (mPGES-1) isomerizes COX-2-derived PGH(2) to PGE(2). Here, we evaluated the effect of mPGES-1-deficiency on the nociceptive behavior in various models of nociception that depend on PGE(2) synthesis. Surprisingly, in the COX-2-dependent zymosan-evoked hyperalgesia model, the nociceptive behavior was not reduced in mPGES-1-deficient mice despite a marked decrease of the spinal PGE(2) synthesis. Similarly, the nociceptive behavior was unaltered in mPGES-1-deficient mice in the formalin test. Importantly, spinal cords and primary spinal cord cells derived from mPGES-1-deficient mice showed a redirection of the PGE(2) synthesis to PGD(2), PGF(2alpha) and 6-keto-PGF(1alpha) (stable metabolite of PGI(2)). Since the latter prostaglandins serve also as mediators of nociception they may compensate the loss of PGE(2) synthesis in mPGES-1-deficient mice.

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mPGES-1 expression and re-direction of eicosanoid metabolism in mPGES-1-deficient peritoneal macrophages. (A) Microsomal fractions (see Supplementary methods for details) of kidney and lung tissues from wild-type and mPGES-1-deficient mice were subjected to western blot analysis to detect relative protein amounts.(B) Peritoneal macrophages were stimulated with 5 μg/ml LPS for 15 hrs and then subjected to western blot analysis to detect relative protein amounts (see Supplementary methods for details). (C) Peritoneal macrophages were stimulated with 5 μg/ml LPS for 15hrs and prostaglandin levels determined from medium by LC-MS/MS. Data are shown as percent of all prostaglandins and are expressed as the mean ± S.E.M.(n= 9).*P <0.05.
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fig01: mPGES-1 expression and re-direction of eicosanoid metabolism in mPGES-1-deficient peritoneal macrophages. (A) Microsomal fractions (see Supplementary methods for details) of kidney and lung tissues from wild-type and mPGES-1-deficient mice were subjected to western blot analysis to detect relative protein amounts.(B) Peritoneal macrophages were stimulated with 5 μg/ml LPS for 15 hrs and then subjected to western blot analysis to detect relative protein amounts (see Supplementary methods for details). (C) Peritoneal macrophages were stimulated with 5 μg/ml LPS for 15hrs and prostaglandin levels determined from medium by LC-MS/MS. Data are shown as percent of all prostaglandins and are expressed as the mean ± S.E.M.(n= 9).*P <0.05.

Mentions: Lack of mPGES-1 expression in mPGES-1 knockout mice was confirmed by western blot analysis of kidney and lung tissue as well as in thioglycollate-elicited LPS-stimulated peritoneal macrophages (Fig. 1A and B). As described previously, the mPGES-1 deficiency did not alter the expression levels of the other PGE2 synthases, namely mPGES-2 and cPGES (Fig. 1B), but caused the re-direction of the eicosanoid synthesis to TxA2 (measured as the stable metabolite TxB2), PGI2 (measured by the stable metabolite 6-keto-PGF1α), PGF2α, and PGD2 (Fig. 1C) [5,7,8]. Importantly, mPGES-1-deficient mice exhibited normal motor function in the pole and the hanging wire test (Supplementary data 1: 1A, B) and normal basal pain thresholds in the tail flick and hot plate tests (supplementary data 1: 1C, D) as compared to wild-type mice.


Consequences of altered eicosanoid patterns for nociceptive processing in mPGES-1-deficient mice.

Brenneis C, Coste O, Schmidt R, Angioni C, Popp L, Nusing RM, Becker W, Scholich K, Geisslinger G - J. Cell. Mol. Med. (2008)

mPGES-1 expression and re-direction of eicosanoid metabolism in mPGES-1-deficient peritoneal macrophages. (A) Microsomal fractions (see Supplementary methods for details) of kidney and lung tissues from wild-type and mPGES-1-deficient mice were subjected to western blot analysis to detect relative protein amounts.(B) Peritoneal macrophages were stimulated with 5 μg/ml LPS for 15 hrs and then subjected to western blot analysis to detect relative protein amounts (see Supplementary methods for details). (C) Peritoneal macrophages were stimulated with 5 μg/ml LPS for 15hrs and prostaglandin levels determined from medium by LC-MS/MS. Data are shown as percent of all prostaglandins and are expressed as the mean ± S.E.M.(n= 9).*P <0.05.
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Related In: Results  -  Collection

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fig01: mPGES-1 expression and re-direction of eicosanoid metabolism in mPGES-1-deficient peritoneal macrophages. (A) Microsomal fractions (see Supplementary methods for details) of kidney and lung tissues from wild-type and mPGES-1-deficient mice were subjected to western blot analysis to detect relative protein amounts.(B) Peritoneal macrophages were stimulated with 5 μg/ml LPS for 15 hrs and then subjected to western blot analysis to detect relative protein amounts (see Supplementary methods for details). (C) Peritoneal macrophages were stimulated with 5 μg/ml LPS for 15hrs and prostaglandin levels determined from medium by LC-MS/MS. Data are shown as percent of all prostaglandins and are expressed as the mean ± S.E.M.(n= 9).*P <0.05.
Mentions: Lack of mPGES-1 expression in mPGES-1 knockout mice was confirmed by western blot analysis of kidney and lung tissue as well as in thioglycollate-elicited LPS-stimulated peritoneal macrophages (Fig. 1A and B). As described previously, the mPGES-1 deficiency did not alter the expression levels of the other PGE2 synthases, namely mPGES-2 and cPGES (Fig. 1B), but caused the re-direction of the eicosanoid synthesis to TxA2 (measured as the stable metabolite TxB2), PGI2 (measured by the stable metabolite 6-keto-PGF1α), PGF2α, and PGD2 (Fig. 1C) [5,7,8]. Importantly, mPGES-1-deficient mice exhibited normal motor function in the pole and the hanging wire test (Supplementary data 1: 1A, B) and normal basal pain thresholds in the tail flick and hot plate tests (supplementary data 1: 1C, D) as compared to wild-type mice.

Bottom Line: Cyclooxygenase-2 (COX-2)-dependent prostaglandin (PG) E(2) synthesis in the spinal cord plays a major role in the development of inflammatory hyperalgesia and allodynia.Microsomal PGE(2) synthase-1 (mPGES-1) isomerizes COX-2-derived PGH(2) to PGE(2).Here, we evaluated the effect of mPGES-1-deficiency on the nociceptive behavior in various models of nociception that depend on PGE(2) synthesis.

View Article: PubMed Central - PubMed

Affiliation: Pharmazentrum Frankfurt, ZAFES, Institut für Klinische Pharmakologie, Klinikum der Johann Wolfgang Goethe-Universität Theodor-Stern-Kai 7, Frankfurt, Germany.

ABSTRACT
Cyclooxygenase-2 (COX-2)-dependent prostaglandin (PG) E(2) synthesis in the spinal cord plays a major role in the development of inflammatory hyperalgesia and allodynia. Microsomal PGE(2) synthase-1 (mPGES-1) isomerizes COX-2-derived PGH(2) to PGE(2). Here, we evaluated the effect of mPGES-1-deficiency on the nociceptive behavior in various models of nociception that depend on PGE(2) synthesis. Surprisingly, in the COX-2-dependent zymosan-evoked hyperalgesia model, the nociceptive behavior was not reduced in mPGES-1-deficient mice despite a marked decrease of the spinal PGE(2) synthesis. Similarly, the nociceptive behavior was unaltered in mPGES-1-deficient mice in the formalin test. Importantly, spinal cords and primary spinal cord cells derived from mPGES-1-deficient mice showed a redirection of the PGE(2) synthesis to PGD(2), PGF(2alpha) and 6-keto-PGF(1alpha) (stable metabolite of PGI(2)). Since the latter prostaglandins serve also as mediators of nociception they may compensate the loss of PGE(2) synthesis in mPGES-1-deficient mice.

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