Gö6976 inhibits LPS-induced microglial TNFα release by suppressing p38 MAP kinase activation
Section snippets
Materials
LPS (Escherichia coli O111:B4) was from List Biological Laboratories (Campbell, CA, USA). Tissue culture plates (24-well) were Costar® from Corning (Corning, NY, USA). Fetal bovine serum, Dulbecco’s modified Eagle’s medium with Ham’s nutrient mixture F12, and genistein were obtained from Life Technologies (Gaithersburg, MD, USA). SB202190, SB202474, Gö6976, Ro31-8220 and H89 were purchased from Calbiochem-Novobiochem International (La Jolla, CA, USA). The mouse TNFα capture antibody, mouse TNFα
Gö6976 and SB202190 inhibited LPS-induced TNFα production by BV2 microglial cells
We examined the effects of protein kinase inhibitors on the increase in production of TNFα, a representative inflammatory factor, in murine microglial BV2 cell cultures stimulated with LPS. TNFα production increased to about 5 ng/ml 3 h after LPS exposure. Pretreatment with the indolocarbazole Gö6976, which protects neurons from endotoxin- or combined cytokine-induced cytotoxicity (Jeohn et al., 2000b), suppressed the LPS-induced increase in TNFα production significantly (Fig. 1A). The
Discussion
Microglial activation in response to such toxins as bacterial LPS, HIV coat protein gp120, and β-amyloid-related peptides leads to the synthesis of inflammation-related products, which have been associated with neuron degeneration in the brain (Dickson et al., 1993, Chao et al., 1995). We propose that Gö6976 may suppress the synthesis of inflammation-related factors in microglia by a PKC-independent mechanism. The results of this study suggest that Gö6976 suppressed LPS-induced TNFα production
Conclusion
Our results indicate that Gö6976 suppressed the phosphorylation of p38 MAPK and consequently, the p38 MAPK activity induced by LPS, and resulted in a decrease in LPS-induced TNFα secretion. The inhibitory function of Gö6976 on the phosphorylation of p38 MAPK was not mediated through the inhibition of any known PKC. Rather, one of the upstream signaling events prior to p38 MAPK, and other than known PKC isotypes, appears to be the functional target of Gö697. Further understanding of the
Acknowledgements
We thank Dr. R.C.C. Chang for valuable discussions and insightful comments on this work and Ms. Crystal Paras for editorial assistance.
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- 1
On sabbatical leave from Division of Science, Truman State University, Kirksville, MO 63501, USA.
- 2
Present address: College of Pharmacy, Kangwon University, Chuncheon, South Korea.