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Propofol attenuates LPS-induced tumor necrosis factor-α, interleukin-6 and nitric oxide expression in canine peripheral blood mononuclear cells possibly through down-regulation of nuclear factor (NF)-κB activation.

Pei Z, Wang J - J. Vet. Med. Sci. (2014)

Bottom Line: Through real-time quantitative PCR and the Griess reagent system, we found that non-cytotoxic levels of propofol significantly inhibited the release of NO and IL-6 and TNF-α gene expression in LPS-induced canine PBMCs.Western blotting revealed that LPS does significantly increase the expression of inducible NO synthase (iNOS) protein in canine PBMCs, while pretreatment with propofol significantly decreases the LPS-induced iNOS protein expression.This diminished TNF-α, IL-6 and iNOS expression, and NO production was in parallel to the respective decreased NF-κB p65 protein nuclear translocation in the LPS-activated canine PBMCs pretreated with 25 µM and 50 µM propofol.

View Article: PubMed Central - PubMed

Affiliation: Department of Veterinary Medicine, College of Animal Sciences, Zhejiang University, Hangzhou 310058, PR China.

ABSTRACT
Sepsis is a major cause of mortality in intensive care medicine. Propofol, an intravenous general anesthetic, has been suggested to have anti-inflammatory properties and able to prevent sepsis induced by Gram-positive and Gram-negative bacteria by down-regulating the gene expression of pro-inflammatory cytokines. However, propofol's anti-inflammatory effects upon canine peripheral blood mononuclear cells (PBMCs) have not yet been clarified. Here, we isolate canine PBMCs and investigate the effects of propofol on the gene expressions of both lipopolysaccharide (LPS)-induced interleukin-6 (IL-6) and tumor necrosis factor (TNF)-α and upon the production of nitric oxide (NO). Through real-time quantitative PCR and the Griess reagent system, we found that non-cytotoxic levels of propofol significantly inhibited the release of NO and IL-6 and TNF-α gene expression in LPS-induced canine PBMCs. Western blotting revealed that LPS does significantly increase the expression of inducible NO synthase (iNOS) protein in canine PBMCs, while pretreatment with propofol significantly decreases the LPS-induced iNOS protein expression. Propofol, at concentration of 25 µM and 50 µM, also significantly inhibited the LPS-induced nuclear translocation of nuclear factor (NF)-κB p65 protein in canine PBMCs. This diminished TNF-α, IL-6 and iNOS expression, and NO production was in parallel to the respective decreased NF-κB p65 protein nuclear translocation in the LPS-activated canine PBMCs pretreated with 25 µM and 50 µM propofol. This suggests that non-cytotoxic levels of propofol pretreatment can down-regulate LPS-induced inflammatory responses in canine PBMCs, possibly by inhibiting the nuclear translocation of the NF-κB p65 protein.

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Propofol inhibits the nuclear translocation of the NF-κB p65 protein in LPS-stimulated canine PBMCs. Canine PBMCs were pretreated with different doses of propofol (0, 25 or 50 µM) for 6 hr and then stimulated with LPS (100 ng/ml) for 1 hr. The NF-κB p65 protein was analyzed by Western blotting. Histone H3 was used as a loading control. Densitometry was normalized to Histone H3 and graphed as the mean ± S.D. *indicates P<0.05 (compared with LPS stimulation alone). Similar results were obtained from three independent experiments.
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fig_004: Propofol inhibits the nuclear translocation of the NF-κB p65 protein in LPS-stimulated canine PBMCs. Canine PBMCs were pretreated with different doses of propofol (0, 25 or 50 µM) for 6 hr and then stimulated with LPS (100 ng/ml) for 1 hr. The NF-κB p65 protein was analyzed by Western blotting. Histone H3 was used as a loading control. Densitometry was normalized to Histone H3 and graphed as the mean ± S.D. *indicates P<0.05 (compared with LPS stimulation alone). Similar results were obtained from three independent experiments.

Mentions: Propofol reduces LPS-induced activation of NF-κB in canine PBMCs: NF-κB activation is essential in the transcriptional regulation of many pro-inflammatory cytokines, including TNF-α and IL-6. The above results indicated that propofol inhibited the expression of TNF-α and IL-6 mRNA in LPS-stimulated canine PBMCs. Therefore, we next investigated whether the upstream NF-κB p65 nuclear translocation was also involved. Using Western blot, we found that the translocation of NF-κB p65 protein into the nucleus was significantly increased in canine PBMCs stimulated with LPS. However, propofol at concentration of 25 µM and 50 µM significantly inhibited the LPS-induced nuclear translocation of the NF-κB p65 protein. The diminished TNF-α, IL-6 and iNOS expression and NO production (Figs. 2 and 3) were in parallel to thedecrease in NF-κB p65 protein nuclear translocation in the LPS-activated canine PBMCs pretreated with 25 µM and 50 µM propofol (Fig. 4Fig. 4.


Propofol attenuates LPS-induced tumor necrosis factor-α, interleukin-6 and nitric oxide expression in canine peripheral blood mononuclear cells possibly through down-regulation of nuclear factor (NF)-κB activation.

Pei Z, Wang J - J. Vet. Med. Sci. (2014)

Propofol inhibits the nuclear translocation of the NF-κB p65 protein in LPS-stimulated canine PBMCs. Canine PBMCs were pretreated with different doses of propofol (0, 25 or 50 µM) for 6 hr and then stimulated with LPS (100 ng/ml) for 1 hr. The NF-κB p65 protein was analyzed by Western blotting. Histone H3 was used as a loading control. Densitometry was normalized to Histone H3 and graphed as the mean ± S.D. *indicates P<0.05 (compared with LPS stimulation alone). Similar results were obtained from three independent experiments.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4363014&req=5

fig_004: Propofol inhibits the nuclear translocation of the NF-κB p65 protein in LPS-stimulated canine PBMCs. Canine PBMCs were pretreated with different doses of propofol (0, 25 or 50 µM) for 6 hr and then stimulated with LPS (100 ng/ml) for 1 hr. The NF-κB p65 protein was analyzed by Western blotting. Histone H3 was used as a loading control. Densitometry was normalized to Histone H3 and graphed as the mean ± S.D. *indicates P<0.05 (compared with LPS stimulation alone). Similar results were obtained from three independent experiments.
Mentions: Propofol reduces LPS-induced activation of NF-κB in canine PBMCs: NF-κB activation is essential in the transcriptional regulation of many pro-inflammatory cytokines, including TNF-α and IL-6. The above results indicated that propofol inhibited the expression of TNF-α and IL-6 mRNA in LPS-stimulated canine PBMCs. Therefore, we next investigated whether the upstream NF-κB p65 nuclear translocation was also involved. Using Western blot, we found that the translocation of NF-κB p65 protein into the nucleus was significantly increased in canine PBMCs stimulated with LPS. However, propofol at concentration of 25 µM and 50 µM significantly inhibited the LPS-induced nuclear translocation of the NF-κB p65 protein. The diminished TNF-α, IL-6 and iNOS expression and NO production (Figs. 2 and 3) were in parallel to thedecrease in NF-κB p65 protein nuclear translocation in the LPS-activated canine PBMCs pretreated with 25 µM and 50 µM propofol (Fig. 4Fig. 4.

Bottom Line: Through real-time quantitative PCR and the Griess reagent system, we found that non-cytotoxic levels of propofol significantly inhibited the release of NO and IL-6 and TNF-α gene expression in LPS-induced canine PBMCs.Western blotting revealed that LPS does significantly increase the expression of inducible NO synthase (iNOS) protein in canine PBMCs, while pretreatment with propofol significantly decreases the LPS-induced iNOS protein expression.This diminished TNF-α, IL-6 and iNOS expression, and NO production was in parallel to the respective decreased NF-κB p65 protein nuclear translocation in the LPS-activated canine PBMCs pretreated with 25 µM and 50 µM propofol.

View Article: PubMed Central - PubMed

Affiliation: Department of Veterinary Medicine, College of Animal Sciences, Zhejiang University, Hangzhou 310058, PR China.

ABSTRACT
Sepsis is a major cause of mortality in intensive care medicine. Propofol, an intravenous general anesthetic, has been suggested to have anti-inflammatory properties and able to prevent sepsis induced by Gram-positive and Gram-negative bacteria by down-regulating the gene expression of pro-inflammatory cytokines. However, propofol's anti-inflammatory effects upon canine peripheral blood mononuclear cells (PBMCs) have not yet been clarified. Here, we isolate canine PBMCs and investigate the effects of propofol on the gene expressions of both lipopolysaccharide (LPS)-induced interleukin-6 (IL-6) and tumor necrosis factor (TNF)-α and upon the production of nitric oxide (NO). Through real-time quantitative PCR and the Griess reagent system, we found that non-cytotoxic levels of propofol significantly inhibited the release of NO and IL-6 and TNF-α gene expression in LPS-induced canine PBMCs. Western blotting revealed that LPS does significantly increase the expression of inducible NO synthase (iNOS) protein in canine PBMCs, while pretreatment with propofol significantly decreases the LPS-induced iNOS protein expression. Propofol, at concentration of 25 µM and 50 µM, also significantly inhibited the LPS-induced nuclear translocation of nuclear factor (NF)-κB p65 protein in canine PBMCs. This diminished TNF-α, IL-6 and iNOS expression, and NO production was in parallel to the respective decreased NF-κB p65 protein nuclear translocation in the LPS-activated canine PBMCs pretreated with 25 µM and 50 µM propofol. This suggests that non-cytotoxic levels of propofol pretreatment can down-regulate LPS-induced inflammatory responses in canine PBMCs, possibly by inhibiting the nuclear translocation of the NF-κB p65 protein.

Show MeSH
Related in: MedlinePlus