Back to Search
Start Over
Glycogen synthase kinase-3 negatively regulates anti-inflammatory interleukin-10 for lipopolysaccharide-induced iNOS/NO biosynthesis and RANTES production in microglial cells
- Source :
- Immunology. 128:e275-e286
- Publication Year :
- 2009
- Publisher :
- Wiley, 2009.
-
Abstract
- The inflammatory effects of glycogen synthase kinase-3 (GSK-3) have been identified; however, the potential mechanism is still controversial. In this study, we investigated the effects of GSK-3-mediated interleukin-10 (IL-10) inhibition on lipopolysaccharide (LPS)-induced inflammation. Treatment with GSK-3 inhibitor significantly blocked LPS-induced nitric oxide (NO) production as well as inducible NO synthase (iNOS) expression in BV2 murine microglial cells and primary rat microglia-enriched cultures. Using an antibody array and enzyme-linked immunosorbent assay, we found that GSK-3-inhibitor treatment blocked LPS-induced upregulation of regulated on activation normal T-cell expressed and secreted (RANTES) and increased IL-10 expression. The time kinetics and dose-response relations were confirmed. Reverse transcription-polymerase chain reaction showed changes on the messenger RNA level as well. Inhibiting GSK-3 using short-interference RNA, and transfecting cells with dominant-negative GSK-3beta, blocked LPS-elicited NO and RANTES expression but increased IL-10 expression. In contrast, GSK-3beta overexpression upregulated NO and RANTES but downregulated IL-10 in LPS-stimulated cells. Treating cells with anti-IL-10 neutralizing antibodies to prevent GSK-3 from downregulating NO and RANTES showed that the anti-inflammatory effects are, at least in part, IL-10-dependent. The involvement of Akt, extracellular signal-regulated kinase, p38 mitogen-activated protein kinase and nuclear factor-kappaB that positively regulated IL-10 was demonstrated. Furthermore, inhibiting GSK-3 increased the nuclear translocation of transcription factors, that all important for IL-10 expression, including CCAAT/enhancer-binding protein beat (C/EBPbeta), C/EBPdelta, cAMP response binding element protein and NF-kappaB. Taken together, these findings reveal that LPS induces iNOS/NO biosynthesis and RANTES production through a mechanism involving GSK-3-mediated IL-10 downregulation.
- Subjects :
- Lipopolysaccharides
p38 mitogen-activated protein kinases
Immunology
Down-Regulation
Nitric Oxide Synthase Type II
Biology
Nitric Oxide
p38 Mitogen-Activated Protein Kinases
Cell Line
Glycogen Synthase Kinase 3
Mice
Downregulation and upregulation
GSK-3
Animals
Immunology and Allergy
RNA, Small Interfering
Protein kinase A
Glycogen synthase
Chemokine CCL5
Protein Kinase Inhibitors
Protein kinase B
Inflammation
Kinase
NF-kappa B
Original Articles
Antibodies, Neutralizing
Molecular biology
Interleukin-10
Rats
Up-Regulation
Interleukin 10
biology.protein
Microglia
Proto-Oncogene Proteins c-akt
Subjects
Details
- ISSN :
- 13652567 and 00192805
- Volume :
- 128
- Database :
- OpenAIRE
- Journal :
- Immunology
- Accession number :
- edsair.doi.dedup.....03af0baf91e27f78b5f52f345e73e946