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Hydrogen sulfide attenuated tumor necrosis factor-α-induced inflammatory signaling and dysfunction in vascular endothelial cells.

Pan LL, Liu XH, Gong QH, Wu D, Zhu YZ - PLoS ONE (2011)

Bottom Line: Herein, we explored the effects and mechanisms of sodium hydrosulfide (NaHS, a H(2)S donor) on tumor necrosis factor (TNF)-α-induced human umbilical vein endothelial cells (HUVEC) dysfunction.Furthermore, TNF-α-induced NF-κB activation assessed by IκBα degradation and p65 phosphorylation and nuclear translocation and ROS production were diminished in cells subjected to treatment with NaHS.H(2)S can exert an anti-inflammatory effect in endothelial cells through a mechanism that involves the up-regulation of HO-1.

View Article: PubMed Central - PubMed

Affiliation: Department of Pharmacology, School of Pharmacy, Fudan University, Shanghai, China.

ABSTRACT

Background: Hydrogen sulfide (H(2)S), the third physiologically relevant gaseous molecule, is recognized increasingly as an anti-inflammatory mediator in various inflammatory conditions. Herein, we explored the effects and mechanisms of sodium hydrosulfide (NaHS, a H(2)S donor) on tumor necrosis factor (TNF)-α-induced human umbilical vein endothelial cells (HUVEC) dysfunction.

Methodology and principal findings: Application of NaHS concentration-dependently suppressed TNF-α-induced mRNA and proteins expressions of intercellular adhesion molecule-1 (ICAM-1) and vascular cell adhesion molecule-1 (VCAM-1), mRNA expression of P-selectin and E-selectin as well as U937 monocytes adhesion to HUVEC. Western blot analysis revealed that the expression of the cytoprotective enzyme, heme oxygenase-1 (HO-1), was induced and coincident with the anti-inflammatory action of NaHS. Furthermore, TNF-α-induced NF-κB activation assessed by IκBα degradation and p65 phosphorylation and nuclear translocation and ROS production were diminished in cells subjected to treatment with NaHS.

Significance: H(2)S can exert an anti-inflammatory effect in endothelial cells through a mechanism that involves the up-regulation of HO-1.

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Related in: MedlinePlus

NaHS upregulated expression of HO-1 in HUVEC.(A) HUVEC were incubated with indicated concentrations of NaHS for 6 hours. Cells were then lysed, and HO-1 expression was analyzed by Western blot. Tubulin was used as loading control. Data represent mean ±S.E.M from 3 independent repeats. *P<0.05, **P<0.01 compared with unstimulated cells. (B) HUVEC were incubated with various concentrations of NaHS for 30 min, cells were then stimulated with TNF-α (10 ng/ml) for 6 h. HO-1 proteins were analyzed by Western blot in HUVEC. Tubulin was used as loading control. NAC concentration was 5 mM. #P<0.05 compared with unstimulated cells, *P<0.05, **P<0.01 compared with TNF-α-stimulated cells. Data are the mean ±S.E.M of results from at least three independent experiments, each performed in duplicate.
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pone-0019766-g004: NaHS upregulated expression of HO-1 in HUVEC.(A) HUVEC were incubated with indicated concentrations of NaHS for 6 hours. Cells were then lysed, and HO-1 expression was analyzed by Western blot. Tubulin was used as loading control. Data represent mean ±S.E.M from 3 independent repeats. *P<0.05, **P<0.01 compared with unstimulated cells. (B) HUVEC were incubated with various concentrations of NaHS for 30 min, cells were then stimulated with TNF-α (10 ng/ml) for 6 h. HO-1 proteins were analyzed by Western blot in HUVEC. Tubulin was used as loading control. NAC concentration was 5 mM. #P<0.05 compared with unstimulated cells, *P<0.05, **P<0.01 compared with TNF-α-stimulated cells. Data are the mean ±S.E.M of results from at least three independent experiments, each performed in duplicate.

Mentions: The effect of NaHS on HO-1 expression was initially explored in cultured HUVEC, Figure 4A shows upon incubation with NaHS (10–100 µM) for 6 hours, a substantial increase in the expression of HO-1 was observed. MTT cell survival assays failed to demonstrate any cellular cytotoxicity at these concentrations (not shown). Thus, we examined the effects of NaHS on HO-1 expression in TNF-α-stimulated HUVEC. As shown in Figure 4B, treatment with TNF-α didn't reveal significant effect on HO-1 expression compared with resting cells. Contrary, NaHS concentration-dependently increased HO-1 expression in TNF-α-stimulated cells. Meanwhile, the upregulation of HO-1 was also observed in NAC-treated cells. Taken together, these results suggested that the expression of HO-1 induced by NaHS may functions as a negative regulator of TNF-α-induced inflammatory responses in HUVEC.


Hydrogen sulfide attenuated tumor necrosis factor-α-induced inflammatory signaling and dysfunction in vascular endothelial cells.

Pan LL, Liu XH, Gong QH, Wu D, Zhu YZ - PLoS ONE (2011)

NaHS upregulated expression of HO-1 in HUVEC.(A) HUVEC were incubated with indicated concentrations of NaHS for 6 hours. Cells were then lysed, and HO-1 expression was analyzed by Western blot. Tubulin was used as loading control. Data represent mean ±S.E.M from 3 independent repeats. *P<0.05, **P<0.01 compared with unstimulated cells. (B) HUVEC were incubated with various concentrations of NaHS for 30 min, cells were then stimulated with TNF-α (10 ng/ml) for 6 h. HO-1 proteins were analyzed by Western blot in HUVEC. Tubulin was used as loading control. NAC concentration was 5 mM. #P<0.05 compared with unstimulated cells, *P<0.05, **P<0.01 compared with TNF-α-stimulated cells. Data are the mean ±S.E.M of results from at least three independent experiments, each performed in duplicate.
© Copyright Policy
Related In: Results  -  Collection

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getmorefigures.php?uid=PMC3091882&req=5

pone-0019766-g004: NaHS upregulated expression of HO-1 in HUVEC.(A) HUVEC were incubated with indicated concentrations of NaHS for 6 hours. Cells were then lysed, and HO-1 expression was analyzed by Western blot. Tubulin was used as loading control. Data represent mean ±S.E.M from 3 independent repeats. *P<0.05, **P<0.01 compared with unstimulated cells. (B) HUVEC were incubated with various concentrations of NaHS for 30 min, cells were then stimulated with TNF-α (10 ng/ml) for 6 h. HO-1 proteins were analyzed by Western blot in HUVEC. Tubulin was used as loading control. NAC concentration was 5 mM. #P<0.05 compared with unstimulated cells, *P<0.05, **P<0.01 compared with TNF-α-stimulated cells. Data are the mean ±S.E.M of results from at least three independent experiments, each performed in duplicate.
Mentions: The effect of NaHS on HO-1 expression was initially explored in cultured HUVEC, Figure 4A shows upon incubation with NaHS (10–100 µM) for 6 hours, a substantial increase in the expression of HO-1 was observed. MTT cell survival assays failed to demonstrate any cellular cytotoxicity at these concentrations (not shown). Thus, we examined the effects of NaHS on HO-1 expression in TNF-α-stimulated HUVEC. As shown in Figure 4B, treatment with TNF-α didn't reveal significant effect on HO-1 expression compared with resting cells. Contrary, NaHS concentration-dependently increased HO-1 expression in TNF-α-stimulated cells. Meanwhile, the upregulation of HO-1 was also observed in NAC-treated cells. Taken together, these results suggested that the expression of HO-1 induced by NaHS may functions as a negative regulator of TNF-α-induced inflammatory responses in HUVEC.

Bottom Line: Herein, we explored the effects and mechanisms of sodium hydrosulfide (NaHS, a H(2)S donor) on tumor necrosis factor (TNF)-α-induced human umbilical vein endothelial cells (HUVEC) dysfunction.Furthermore, TNF-α-induced NF-κB activation assessed by IκBα degradation and p65 phosphorylation and nuclear translocation and ROS production were diminished in cells subjected to treatment with NaHS.H(2)S can exert an anti-inflammatory effect in endothelial cells through a mechanism that involves the up-regulation of HO-1.

View Article: PubMed Central - PubMed

Affiliation: Department of Pharmacology, School of Pharmacy, Fudan University, Shanghai, China.

ABSTRACT

Background: Hydrogen sulfide (H(2)S), the third physiologically relevant gaseous molecule, is recognized increasingly as an anti-inflammatory mediator in various inflammatory conditions. Herein, we explored the effects and mechanisms of sodium hydrosulfide (NaHS, a H(2)S donor) on tumor necrosis factor (TNF)-α-induced human umbilical vein endothelial cells (HUVEC) dysfunction.

Methodology and principal findings: Application of NaHS concentration-dependently suppressed TNF-α-induced mRNA and proteins expressions of intercellular adhesion molecule-1 (ICAM-1) and vascular cell adhesion molecule-1 (VCAM-1), mRNA expression of P-selectin and E-selectin as well as U937 monocytes adhesion to HUVEC. Western blot analysis revealed that the expression of the cytoprotective enzyme, heme oxygenase-1 (HO-1), was induced and coincident with the anti-inflammatory action of NaHS. Furthermore, TNF-α-induced NF-κB activation assessed by IκBα degradation and p65 phosphorylation and nuclear translocation and ROS production were diminished in cells subjected to treatment with NaHS.

Significance: H(2)S can exert an anti-inflammatory effect in endothelial cells through a mechanism that involves the up-regulation of HO-1.

Show MeSH
Related in: MedlinePlus