Abstract
Interferon-γ (IFN-γ) plays a crucial role in innate immunity and inflammation. It causes the synergistic effect on endotoxin lipopolysaccharide (LPS)-stimulated inducible nitric oxide synthase (iNOS)/NO biosynthesis; however, the mechanism remains unclear. In the present study, we investigated the effects of glycogen synthase kinase-3 (GSK-3)-mediated inhibition of anti-inflammatory interleukin-10 (IL-10). We found, in LPS-stimulated macrophages, that IFN-γ increased iNOS expression and NO production in a time-dependent manner. In addition, ELISA analysis showed the upregulation of tumor necrosis factor-α and regulated on activation, normal T expressed and secreted, and the downregulation of IL-10. RT-PCR further showed changes in the IL-10 mRNA level as well. Treating cells with recombinant IL-10 showed a decrease in IFN-γ/LPS-induced iNOS/NO biosynthesis, whereas anti-IL-10 neutralizing antibodies enhanced this effect, suggesting that IL-10 acts in an anti-inflammatory role. GSK-3-inhibitor treatment blocked IFN-γ/LPS-induced iNOS/NO biosynthesis but upregulated IL-10 production. Inhibiting GSK-3 using short-interference RNA showed similar results. Additionally, treating cells with anti-IL-10 neutralizing antibodies blocked these effects. We further showed that inhibiting GSK-3 increased phosphorylation of transcription factor cyclic AMP response element binding protein. Inhibiting protein tyrosine kinase Pyk2, an upstream regulator of GSK-3β, caused inhibition on IFN-γ/LPS-induced GSK-3β phosphorylation at tyrosine 216 and iNOS/NO biosynthesis. Taken together, these findings reveal the involvement of GSK-3-inhibited IL-10 on the induction of iNOS/NO biosynthesis by IFN-γ synergized with LPS.
| Original language | English |
|---|---|
| Pages (from-to) | 746-755 |
| Number of pages | 10 |
| Journal | Journal of Cellular Biochemistry |
| Volume | 105 |
| Issue number | 3 |
| DOIs | |
| Publication status | Published - Oct 15 2008 |
| Externally published | Yes |
Fingerprint
Keywords
- GSK-3
- IFN-γ
- IL-10
- iNOS
- LPS
- Macrophage
- NO
ASJC Scopus subject areas
- Biochemistry
- Cell Biology
- Molecular Biology
Cite this
IFN-γ synergizes with LPS to induce nitric oxide biosynthesis through glycogen synthase kinase-3-inhibited IL-10. / Lin, Chiou Feng; Tsai, Cheng Chieh; Huang, Wei Ching; Wang, Chi Yun; Tseng, Hsiang Chi; Wang, Yi; Kai, Jui In; Wang, Szu Wen; Cheng, Yi Lin.
In: Journal of Cellular Biochemistry, Vol. 105, No. 3, 15.10.2008, p. 746-755.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - IFN-γ synergizes with LPS to induce nitric oxide biosynthesis through glycogen synthase kinase-3-inhibited IL-10
AU - Lin, Chiou Feng
AU - Tsai, Cheng Chieh
AU - Huang, Wei Ching
AU - Wang, Chi Yun
AU - Tseng, Hsiang Chi
AU - Wang, Yi
AU - Kai, Jui In
AU - Wang, Szu Wen
AU - Cheng, Yi Lin
PY - 2008/10/15
Y1 - 2008/10/15
N2 - Interferon-γ (IFN-γ) plays a crucial role in innate immunity and inflammation. It causes the synergistic effect on endotoxin lipopolysaccharide (LPS)-stimulated inducible nitric oxide synthase (iNOS)/NO biosynthesis; however, the mechanism remains unclear. In the present study, we investigated the effects of glycogen synthase kinase-3 (GSK-3)-mediated inhibition of anti-inflammatory interleukin-10 (IL-10). We found, in LPS-stimulated macrophages, that IFN-γ increased iNOS expression and NO production in a time-dependent manner. In addition, ELISA analysis showed the upregulation of tumor necrosis factor-α and regulated on activation, normal T expressed and secreted, and the downregulation of IL-10. RT-PCR further showed changes in the IL-10 mRNA level as well. Treating cells with recombinant IL-10 showed a decrease in IFN-γ/LPS-induced iNOS/NO biosynthesis, whereas anti-IL-10 neutralizing antibodies enhanced this effect, suggesting that IL-10 acts in an anti-inflammatory role. GSK-3-inhibitor treatment blocked IFN-γ/LPS-induced iNOS/NO biosynthesis but upregulated IL-10 production. Inhibiting GSK-3 using short-interference RNA showed similar results. Additionally, treating cells with anti-IL-10 neutralizing antibodies blocked these effects. We further showed that inhibiting GSK-3 increased phosphorylation of transcription factor cyclic AMP response element binding protein. Inhibiting protein tyrosine kinase Pyk2, an upstream regulator of GSK-3β, caused inhibition on IFN-γ/LPS-induced GSK-3β phosphorylation at tyrosine 216 and iNOS/NO biosynthesis. Taken together, these findings reveal the involvement of GSK-3-inhibited IL-10 on the induction of iNOS/NO biosynthesis by IFN-γ synergized with LPS.
AB - Interferon-γ (IFN-γ) plays a crucial role in innate immunity and inflammation. It causes the synergistic effect on endotoxin lipopolysaccharide (LPS)-stimulated inducible nitric oxide synthase (iNOS)/NO biosynthesis; however, the mechanism remains unclear. In the present study, we investigated the effects of glycogen synthase kinase-3 (GSK-3)-mediated inhibition of anti-inflammatory interleukin-10 (IL-10). We found, in LPS-stimulated macrophages, that IFN-γ increased iNOS expression and NO production in a time-dependent manner. In addition, ELISA analysis showed the upregulation of tumor necrosis factor-α and regulated on activation, normal T expressed and secreted, and the downregulation of IL-10. RT-PCR further showed changes in the IL-10 mRNA level as well. Treating cells with recombinant IL-10 showed a decrease in IFN-γ/LPS-induced iNOS/NO biosynthesis, whereas anti-IL-10 neutralizing antibodies enhanced this effect, suggesting that IL-10 acts in an anti-inflammatory role. GSK-3-inhibitor treatment blocked IFN-γ/LPS-induced iNOS/NO biosynthesis but upregulated IL-10 production. Inhibiting GSK-3 using short-interference RNA showed similar results. Additionally, treating cells with anti-IL-10 neutralizing antibodies blocked these effects. We further showed that inhibiting GSK-3 increased phosphorylation of transcription factor cyclic AMP response element binding protein. Inhibiting protein tyrosine kinase Pyk2, an upstream regulator of GSK-3β, caused inhibition on IFN-γ/LPS-induced GSK-3β phosphorylation at tyrosine 216 and iNOS/NO biosynthesis. Taken together, these findings reveal the involvement of GSK-3-inhibited IL-10 on the induction of iNOS/NO biosynthesis by IFN-γ synergized with LPS.
KW - GSK-3
KW - IFN-γ
KW - IL-10
KW - iNOS
KW - LPS
KW - Macrophage
KW - NO
UR - http://www.scopus.com/inward/record.url?scp=54849436053&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=54849436053&partnerID=8YFLogxK
U2 - 10.1002/jcb.21868
DO - 10.1002/jcb.21868
M3 - Article
C2 - 18655171
AN - SCOPUS:54849436053
VL - 105
SP - 746
EP - 755
JO - Journal of Cellular Biochemistry
JF - Journal of Cellular Biochemistry
SN - 0730-2312
IS - 3
ER -