Intravenous anesthetic propofol binds to 5‐lipoxygenase and attenuates leukotriene B₄production

Abstract

Propofol is an intravenous anesthetic that produces its anesthetic effect, largely the GABA receptor in the CNS, and also reduces the -formyl-methionyl-leucyl-phenylalanine (fMLP)-induced neutrophil respiratory burst. Because fMLP-stimulated neutrophils produce leukotriene (LT)B, we examined the effect of propofol on LTB production and Cecal ligation and puncture surgery was performed in mice, with or without exposure to propofol. Propofol attenuated the production of 5-lipoxygenase (5-LOX)-related arachidonic acid (AA) derivatives in the peritoneal fluid. Also, in the experiments on fMLP-stimulated neutrophils and 5-LOX-transfected human embryonic kidney cells, propofol attenuated the production of 5-LOX-related AA derivatives. Based on these results, we hypothesized that propofol would directly affect 5-LOX function. Using-azi-propofol (Azi), we photolabeled stable 5-LOX protein, which had been used to solve the X-ray crystallographic structure of 5-LOX, and examined the binding site(s) of propofol on 5-LOX. Two propofol binding pockets were identified near the active site of 5-LOX. Alanine scanning mutagenesis was performed for the residues of 5-LOX in the vicinity of propofol, and we evaluated the functional role of these pockets in LTB production. We demonstrated that these pockets were functionally important for 5-LOX activity. These two pockets can be used to explore a novel 5-LOX inhibitor in the future.-Okuno, T., Koutsogiannaki, S., Ohba, M., Chamberlain, M., Bu, W., Lin, F.-Y., Eckenhoff, R. G., Yokomizo T., Yuki, K. Intravenous anesthetic propofol binds to 5-lipoxygenase and attenuates leukotriene B production.

The effect of anesthetics on toll like receptor 9

“…Azi‐isoflurane, azi‐sevoflurane, and m‐azipropofol (AziPm) are photoactivatable analogues of isoflurane, sevoflurane and propofol, respectively 22,24‐26 . Azi‐isoflurane (30 µM), azi‐sevoflurane (50 µM), or aziPm (10 µM) was equilibrated with TLR9 protein (1 mg/mL) in a reaction volume of 300 µL for 10 minutes.…”

“…Azi-isoflurane, azi-sevoflurane, and m-azipropofol (AziPm) are photoactivatable analogues of isoflurane, sevoflurane and propofol, respectively. 22,[24][25][26] Aziisoflurane (30 µM), azi-sevoflurane (50 µM), or aziPm (10 µM) was equilibrated with TLR9 protein (1 mg/mL) in a reaction volume of 300 µL for 10 minutes. Samples equilibrated with azi-isoflurane or azi-sevoflurane were exposed to 300 nm light using Rayonet RPR-3000 lamp (Southern New England Ultraviolet, Branford, CT, USA).…”

The volatile anesthetic sevoflurane reduces neutrophil apoptosis via Fas death domain–Fas‐associated death domain interaction

“…Reverse‐phase mass spectrometry‐based quantitation technique was used for lipoxin A 4 (LXA 4 ) and maresin 1 (MaR1) measurement as previously described by Okuno et al . (27). The samples were diluted with 2 ml of methanol and 7 ml of water containing 0.1% formic acid containing a mixture of deuteriumlabeled eicosanoids as internal standards, and then loaded on Oasis HLB cartridge (Waters, Milford, MA, USA).…”

Volatile anesthetics isoflurane and sevoflurane directly target and attenuate Toll‐like receptor 4 system

“…Reverse-phase mass spectrometry (MS)-based quantitation technique for eicosanoids was previously described by Okuno et al (9). The lipids were extracted with methanol and diluted with water containing 0.1% formic acid to yield a final methanol concentration of 20%.…”

“…1C). Previously we have shown that intravenous anesthetic propofol directly bound to and attenuated 5-LOX (9). Thus, we tested if isoflurane also directly bound to 5-LOX using photoactivatable isoflurane (azi-isoflurane).…”