Biphasic modulation of NOS expression, protein and nitrite products by hydroxocobalamin underlies its protective effect in endotoxemic shock: downstream regulation of COX-2, IL-1β, TNF-α, IL-6, and HMGB1 expression

André L F Sampaio; Jesmond Dalli; Vincenzo Brancaleone; Fulvio D'Acquisto; Mauro Perretti; Carmen Wheatley

doi:10.1155/2013/741804

Biphasic modulation of NOS expression, protein and nitrite products by hydroxocobalamin underlies its protective effect in endotoxemic shock: downstream regulation of COX-2, IL-1β, TNF-α, IL-6, and HMGB1 expression

André L F Sampaio, Jesmond Dalli, Vincenzo Brancaleone, Fulvio D'Acquisto, Mauro Perretti, Carmen Wheatley

Research output: Contribution to journal › Article › peer-review

Abstract

BACKGROUND: NOS/•NO inhibitors are potential therapeutics for sepsis, yet they increase clinical mortality. However, there has been no in vivo investigation of the (in vitro) •NO scavenger, cobalamin's (Cbl) endogenous effects on NOS/•NO/inflammatory mediators during the immune response to sepsis.

METHODS: We used quantitative polymerase chain reaction (qPCR), ELISA, Western blot, and NOS Griess assays, in a C57BL/6 mouse, acute endotoxaemia model.

RESULTS: During the immune response, pro-inflammatory phase, parenteral hydroxocobalamin (HOCbl) treatment partially inhibits hepatic, but not lung, iNOS mRNA and promotes lung eNOS mRNA, but attenuates the LPS hepatic rise in eNOS mRNA, whilst paradoxically promoting high iNOS/eNOS protein translation, but relatively moderate •NO production. HOCbl/NOS/•NO regulation is reciprocally associated with lower 4 h expression of TNF-α, IL-1β, COX-2, and lower circulating TNF-α, but not IL-6. In resolution, 24 h after LPS, HOCbl completely abrogates a major late mediator of sepsis mortality, high mobility group box 1 (HMGB1) mRNA, inhibits iNOS mRNA, and attenuates LPS-induced hepatic inhibition of eNOS mRNA, whilst showing increased, but still moderate, NOS activity, relative to LPS only. experiments (LPS+D-Galactosamine) HOCbl afforded significant, dose-dependent protection in mice.

CONCLUSIONS: HOCbl produces a complex, time- and organ-dependent, selective regulation of NOS/•NO during endotoxaemia, corollary regulation of downstream inflammatory mediators, and increased survival. This merits clinical evaluation.

Original language	English
Pages (from-to)	741804
Journal	Mediators of inflammation
Volume	2013
DOIs	https://doi.org/10.1155/2013/741804
Publication status	Published - 2013

Keywords

Animals
Blotting, Western
Cyclooxygenase 2
Endotoxemia
Galactosamine
HMGB1 Protein
Hydroxocobalamin
Interleukin-1beta
Interleukin-6
Liver
Lung
Male
Mice
Mice, Inbred C57BL
Nitric Oxide
Nitric Oxide Synthase Type II
Nitric Oxide Synthase Type III
Reverse Transcriptase Polymerase Chain Reaction
Tumor Necrosis Factor-alpha
Journal Article
Research Support, Non-U.S. Gov't

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10.1155/2013/741804

Cite this

Sampaio, A. L. F., Dalli, J., Brancaleone, V., D'Acquisto, F., Perretti, M., & Wheatley, C. (2013). Biphasic modulation of NOS expression, protein and nitrite products by hydroxocobalamin underlies its protective effect in endotoxemic shock: downstream regulation of COX-2, IL-1β, TNF-α, IL-6, and HMGB1 expression. Mediators of inflammation, 2013, 741804. https://doi.org/10.1155/2013/741804

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title = "Biphasic modulation of NOS expression, protein and nitrite products by hydroxocobalamin underlies its protective effect in endotoxemic shock: downstream regulation of COX-2, IL-1β, TNF-α, IL-6, and HMGB1 expression",

abstract = "BACKGROUND: NOS/•NO inhibitors are potential therapeutics for sepsis, yet they increase clinical mortality. However, there has been no in vivo investigation of the (in vitro) •NO scavenger, cobalamin's (Cbl) endogenous effects on NOS/•NO/inflammatory mediators during the immune response to sepsis.METHODS: We used quantitative polymerase chain reaction (qPCR), ELISA, Western blot, and NOS Griess assays, in a C57BL/6 mouse, acute endotoxaemia model.RESULTS: During the immune response, pro-inflammatory phase, parenteral hydroxocobalamin (HOCbl) treatment partially inhibits hepatic, but not lung, iNOS mRNA and promotes lung eNOS mRNA, but attenuates the LPS hepatic rise in eNOS mRNA, whilst paradoxically promoting high iNOS/eNOS protein translation, but relatively moderate •NO production. HOCbl/NOS/•NO regulation is reciprocally associated with lower 4 h expression of TNF-α, IL-1β, COX-2, and lower circulating TNF-α, but not IL-6. In resolution, 24 h after LPS, HOCbl completely abrogates a major late mediator of sepsis mortality, high mobility group box 1 (HMGB1) mRNA, inhibits iNOS mRNA, and attenuates LPS-induced hepatic inhibition of eNOS mRNA, whilst showing increased, but still moderate, NOS activity, relative to LPS only. experiments (LPS+D-Galactosamine) HOCbl afforded significant, dose-dependent protection in mice.CONCLUSIONS: HOCbl produces a complex, time- and organ-dependent, selective regulation of NOS/•NO during endotoxaemia, corollary regulation of downstream inflammatory mediators, and increased survival. This merits clinical evaluation.",

keywords = "Animals, Blotting, Western, Cyclooxygenase 2, Endotoxemia, Galactosamine, HMGB1 Protein, Hydroxocobalamin, Interleukin-1beta, Interleukin-6, Liver, Lung, Male, Mice, Mice, Inbred C57BL, Nitric Oxide, Nitric Oxide Synthase Type II, Nitric Oxide Synthase Type III, Reverse Transcriptase Polymerase Chain Reaction, Tumor Necrosis Factor-alpha, Journal Article, Research Support, Non-U.S. Gov't",

author = "Sampaio, {Andr{\'e} L F} and Jesmond Dalli and Vincenzo Brancaleone and Fulvio D'Acquisto and Mauro Perretti and Carmen Wheatley",

year = "2013",

doi = "10.1155/2013/741804",

language = "English",

volume = "2013",

pages = "741804",

journal = "Mediators of inflammation",

issn = "0962-9351",

publisher = "Hindawi Publishing Corporation",

}

Sampaio, ALF, Dalli, J, Brancaleone, V, D'Acquisto, F, Perretti, M & Wheatley, C 2013, 'Biphasic modulation of NOS expression, protein and nitrite products by hydroxocobalamin underlies its protective effect in endotoxemic shock: downstream regulation of COX-2, IL-1β, TNF-α, IL-6, and HMGB1 expression', Mediators of inflammation, vol. 2013, pp. 741804. https://doi.org/10.1155/2013/741804

Biphasic modulation of NOS expression, protein and nitrite products by hydroxocobalamin underlies its protective effect in endotoxemic shock: downstream regulation of COX-2, IL-1β, TNF-α, IL-6, and HMGB1 expression. / Sampaio, André L F; Dalli, Jesmond; Brancaleone, Vincenzo et al.
In: Mediators of inflammation, Vol. 2013, 2013, p. 741804.

Research output: Contribution to journal › Article › peer-review

TY - JOUR

T1 - Biphasic modulation of NOS expression, protein and nitrite products by hydroxocobalamin underlies its protective effect in endotoxemic shock

T2 - downstream regulation of COX-2, IL-1β, TNF-α, IL-6, and HMGB1 expression

AU - Sampaio, André L F

AU - Dalli, Jesmond

AU - Brancaleone, Vincenzo

AU - D'Acquisto, Fulvio

AU - Perretti, Mauro

AU - Wheatley, Carmen

PY - 2013

Y1 - 2013

N2 - BACKGROUND: NOS/•NO inhibitors are potential therapeutics for sepsis, yet they increase clinical mortality. However, there has been no in vivo investigation of the (in vitro) •NO scavenger, cobalamin's (Cbl) endogenous effects on NOS/•NO/inflammatory mediators during the immune response to sepsis.METHODS: We used quantitative polymerase chain reaction (qPCR), ELISA, Western blot, and NOS Griess assays, in a C57BL/6 mouse, acute endotoxaemia model.RESULTS: During the immune response, pro-inflammatory phase, parenteral hydroxocobalamin (HOCbl) treatment partially inhibits hepatic, but not lung, iNOS mRNA and promotes lung eNOS mRNA, but attenuates the LPS hepatic rise in eNOS mRNA, whilst paradoxically promoting high iNOS/eNOS protein translation, but relatively moderate •NO production. HOCbl/NOS/•NO regulation is reciprocally associated with lower 4 h expression of TNF-α, IL-1β, COX-2, and lower circulating TNF-α, but not IL-6. In resolution, 24 h after LPS, HOCbl completely abrogates a major late mediator of sepsis mortality, high mobility group box 1 (HMGB1) mRNA, inhibits iNOS mRNA, and attenuates LPS-induced hepatic inhibition of eNOS mRNA, whilst showing increased, but still moderate, NOS activity, relative to LPS only. experiments (LPS+D-Galactosamine) HOCbl afforded significant, dose-dependent protection in mice.CONCLUSIONS: HOCbl produces a complex, time- and organ-dependent, selective regulation of NOS/•NO during endotoxaemia, corollary regulation of downstream inflammatory mediators, and increased survival. This merits clinical evaluation.

AB - BACKGROUND: NOS/•NO inhibitors are potential therapeutics for sepsis, yet they increase clinical mortality. However, there has been no in vivo investigation of the (in vitro) •NO scavenger, cobalamin's (Cbl) endogenous effects on NOS/•NO/inflammatory mediators during the immune response to sepsis.METHODS: We used quantitative polymerase chain reaction (qPCR), ELISA, Western blot, and NOS Griess assays, in a C57BL/6 mouse, acute endotoxaemia model.RESULTS: During the immune response, pro-inflammatory phase, parenteral hydroxocobalamin (HOCbl) treatment partially inhibits hepatic, but not lung, iNOS mRNA and promotes lung eNOS mRNA, but attenuates the LPS hepatic rise in eNOS mRNA, whilst paradoxically promoting high iNOS/eNOS protein translation, but relatively moderate •NO production. HOCbl/NOS/•NO regulation is reciprocally associated with lower 4 h expression of TNF-α, IL-1β, COX-2, and lower circulating TNF-α, but not IL-6. In resolution, 24 h after LPS, HOCbl completely abrogates a major late mediator of sepsis mortality, high mobility group box 1 (HMGB1) mRNA, inhibits iNOS mRNA, and attenuates LPS-induced hepatic inhibition of eNOS mRNA, whilst showing increased, but still moderate, NOS activity, relative to LPS only. experiments (LPS+D-Galactosamine) HOCbl afforded significant, dose-dependent protection in mice.CONCLUSIONS: HOCbl produces a complex, time- and organ-dependent, selective regulation of NOS/•NO during endotoxaemia, corollary regulation of downstream inflammatory mediators, and increased survival. This merits clinical evaluation.

KW - Animals

KW - Blotting, Western

KW - Cyclooxygenase 2

KW - Endotoxemia

KW - Galactosamine

KW - HMGB1 Protein

KW - Hydroxocobalamin

KW - Interleukin-1beta

KW - Interleukin-6

KW - Liver

KW - Lung

KW - Male

KW - Mice

KW - Mice, Inbred C57BL

KW - Nitric Oxide

KW - Nitric Oxide Synthase Type II

KW - Nitric Oxide Synthase Type III

KW - Reverse Transcriptase Polymerase Chain Reaction

KW - Tumor Necrosis Factor-alpha

KW - Journal Article

KW - Research Support, Non-U.S. Gov't

U2 - 10.1155/2013/741804

DO - 10.1155/2013/741804

M3 - Article

C2 - 23781123

SN - 0962-9351

VL - 2013

SP - 741804

JO - Mediators of inflammation

JF - Mediators of inflammation

ER -