Limits...
Anti-inflammatory effects of α-galactosylceramide analogs in activated microglia: involvement of the p38 MAPK signaling pathway.

Jeong YH, Kim Y, Song H, Chung YS, Park SB, Kim HS - PLoS ONE (2014)

Bottom Line: Here, we designed and synthesized α-galactosylceramide (α-GalCer) analogs to exert anti-inflammatory effects in activated microglia.After identification of 4d and 4e as hit compounds, we further investigated the underlying mechanism of their anti-inflammatory effects using RT-PCR analysis.Taken together, these results suggest that p38 MAPK plays an important role in the anti-inflammatory effects of 4d and 4e via the modulation of NF-κB and AP-1 activities.

View Article: PubMed Central - PubMed

Affiliation: Department of Molecular Medicine and Global Top5 Research Program, Tissue Injury Defense Research Center, Ewha Womans University Medical School, Seoul, Republic of Korea.

ABSTRACT
Microglial activation plays a pivotal role in the development and progression of neurodegenerative diseases. Thus, anti-inflammatory agents that control microglial activation can serve as potential therapeutic agents for neurodegenerative diseases. Here, we designed and synthesized α-galactosylceramide (α-GalCer) analogs to exert anti-inflammatory effects in activated microglia. We performed biological evaluations of 25 α-GalCer analogs and observed an interesting preliminary structure-activity relationship in their inhibitory influence on NO release and TNF-α production in LPS-stimulated BV2 microglial cells. After identification of 4d and 4e as hit compounds, we further investigated the underlying mechanism of their anti-inflammatory effects using RT-PCR analysis. We confirmed that 4d and 4e regulate the expression of iNOS, COX-2, IL-1β, and IL-6 at the mRNA level and the expression of TNF-α at the post-transcriptional level. In addition, both 4d and 4e inhibited LPS-induced DNA binding activities of NF-κB and AP-1 and phosphorylation of p38 MAPK without affecting other MAP kinases. When we examined the anti-inflammatory effect of a p38 MAPK-specific inhibitor, SB203580, on microglial activation, we observed an identical inhibitory pattern as that of 4d and 4e, not only on NO and TNF-α production but also on the DNA binding activities of NF-κB and AP-1. Taken together, these results suggest that p38 MAPK plays an important role in the anti-inflammatory effects of 4d and 4e via the modulation of NF-κB and AP-1 activities.

Show MeSH

Related in: MedlinePlus

Inhibitory effects of 4d and 4e on NO and TNF-α production in LTA- or Poly I∶C-stimulated BV2 microglial cells.Cells were incubated for 16 h with LTA (10 µg/mL) (A) or Poly I∶C (10 µg/mL) (B) in the absence or presence of α-GalCer analogs 4d and 4e (5 µM), and the amounts of released NO and TNF-α were measured in supernatants. Treatment with α-GalCer analogs alone did not affect NO or TNF-α production. Bars indicate the mean ± S.E.M. of three independent experiments. *P<0.05; significantly different from stimulated microglial cells.
© Copyright Policy
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC3921125&req=5

pone-0087030-g008: Inhibitory effects of 4d and 4e on NO and TNF-α production in LTA- or Poly I∶C-stimulated BV2 microglial cells.Cells were incubated for 16 h with LTA (10 µg/mL) (A) or Poly I∶C (10 µg/mL) (B) in the absence or presence of α-GalCer analogs 4d and 4e (5 µM), and the amounts of released NO and TNF-α were measured in supernatants. Treatment with α-GalCer analogs alone did not affect NO or TNF-α production. Bars indicate the mean ± S.E.M. of three independent experiments. *P<0.05; significantly different from stimulated microglial cells.

Mentions: To demonstrate that anti-inflammatory effects of 4d and 4e are not confined to LPS stimulation, we examined the inhibitory effect of 4d and 4e in other agonist-stimulated conditions. As shown in Figure 8A and 8B, NO release and TNF-α production in lipoteichoic acid (LTA, TLR2 agonist)- [29], [30] or PolyI∶C (TLR3 agonist)-stimulated BV2 cells [31], [32] were also inhibited upon treatment with 4d and 4e, confirming that 4d and 4e exert anti-inflammatory effects in microglia under various inflammatory conditions.


Anti-inflammatory effects of α-galactosylceramide analogs in activated microglia: involvement of the p38 MAPK signaling pathway.

Jeong YH, Kim Y, Song H, Chung YS, Park SB, Kim HS - PLoS ONE (2014)

Inhibitory effects of 4d and 4e on NO and TNF-α production in LTA- or Poly I∶C-stimulated BV2 microglial cells.Cells were incubated for 16 h with LTA (10 µg/mL) (A) or Poly I∶C (10 µg/mL) (B) in the absence or presence of α-GalCer analogs 4d and 4e (5 µM), and the amounts of released NO and TNF-α were measured in supernatants. Treatment with α-GalCer analogs alone did not affect NO or TNF-α production. Bars indicate the mean ± S.E.M. of three independent experiments. *P<0.05; significantly different from stimulated microglial cells.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC3921125&req=5

pone-0087030-g008: Inhibitory effects of 4d and 4e on NO and TNF-α production in LTA- or Poly I∶C-stimulated BV2 microglial cells.Cells were incubated for 16 h with LTA (10 µg/mL) (A) or Poly I∶C (10 µg/mL) (B) in the absence or presence of α-GalCer analogs 4d and 4e (5 µM), and the amounts of released NO and TNF-α were measured in supernatants. Treatment with α-GalCer analogs alone did not affect NO or TNF-α production. Bars indicate the mean ± S.E.M. of three independent experiments. *P<0.05; significantly different from stimulated microglial cells.
Mentions: To demonstrate that anti-inflammatory effects of 4d and 4e are not confined to LPS stimulation, we examined the inhibitory effect of 4d and 4e in other agonist-stimulated conditions. As shown in Figure 8A and 8B, NO release and TNF-α production in lipoteichoic acid (LTA, TLR2 agonist)- [29], [30] or PolyI∶C (TLR3 agonist)-stimulated BV2 cells [31], [32] were also inhibited upon treatment with 4d and 4e, confirming that 4d and 4e exert anti-inflammatory effects in microglia under various inflammatory conditions.

Bottom Line: Here, we designed and synthesized α-galactosylceramide (α-GalCer) analogs to exert anti-inflammatory effects in activated microglia.After identification of 4d and 4e as hit compounds, we further investigated the underlying mechanism of their anti-inflammatory effects using RT-PCR analysis.Taken together, these results suggest that p38 MAPK plays an important role in the anti-inflammatory effects of 4d and 4e via the modulation of NF-κB and AP-1 activities.

View Article: PubMed Central - PubMed

Affiliation: Department of Molecular Medicine and Global Top5 Research Program, Tissue Injury Defense Research Center, Ewha Womans University Medical School, Seoul, Republic of Korea.

ABSTRACT
Microglial activation plays a pivotal role in the development and progression of neurodegenerative diseases. Thus, anti-inflammatory agents that control microglial activation can serve as potential therapeutic agents for neurodegenerative diseases. Here, we designed and synthesized α-galactosylceramide (α-GalCer) analogs to exert anti-inflammatory effects in activated microglia. We performed biological evaluations of 25 α-GalCer analogs and observed an interesting preliminary structure-activity relationship in their inhibitory influence on NO release and TNF-α production in LPS-stimulated BV2 microglial cells. After identification of 4d and 4e as hit compounds, we further investigated the underlying mechanism of their anti-inflammatory effects using RT-PCR analysis. We confirmed that 4d and 4e regulate the expression of iNOS, COX-2, IL-1β, and IL-6 at the mRNA level and the expression of TNF-α at the post-transcriptional level. In addition, both 4d and 4e inhibited LPS-induced DNA binding activities of NF-κB and AP-1 and phosphorylation of p38 MAPK without affecting other MAP kinases. When we examined the anti-inflammatory effect of a p38 MAPK-specific inhibitor, SB203580, on microglial activation, we observed an identical inhibitory pattern as that of 4d and 4e, not only on NO and TNF-α production but also on the DNA binding activities of NF-κB and AP-1. Taken together, these results suggest that p38 MAPK plays an important role in the anti-inflammatory effects of 4d and 4e via the modulation of NF-κB and AP-1 activities.

Show MeSH
Related in: MedlinePlus