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Angiogenin reduces immune inflammation via inhibition of TANK-binding kinase 1 expression in human corneal fibroblast cells.

Lee SH, Kim KW, Min KM, Kim KW, Chang SI, Kim JC - Mediators Inflamm. (2014)

Bottom Line: Although ANG may be involved in the anti-inflammatory response, there is no evidence that it has direct anti-inflammatory effects.These findings suggest that ANG suppressed the TNF-α-induced inflammatory response in HCFs through inhibition of TBK1-mediated NF-κB nuclear translocation.These novel results are likely to play a significant role in the selection of immune-mediated inflammatory therapeutic targets and may shed light on the pathogenesis of immune-mediated inflammatory diseases.

View Article: PubMed Central - PubMed

Affiliation: Department of Ophthalmology, College of Medicine, Chung-Ang University Hospital, 224-1 Heukseok-dong, Dongjak-Gu, Seoul 156-755, Republic of Korea ; Department of Medicine, Graduate School, Chung-Ang University, Seoul, Republic of Korea.

ABSTRACT
Angiogenin (ANG) is reportedly multifunctional, with roles in angiogenesis and autoimmune diseases. This protein is involved in the innate immune system and has been implicated in several inflammatory diseases. Although ANG may be involved in the anti-inflammatory response, there is no evidence that it has direct anti-inflammatory effects. In this study we sought to determine whether ANG has an anti-inflammatory effect in human corneal fibroblasts (HCFs) exposed to media containing tumor necrosis factor-alpha (TNF-α). We found that ANG reduced the mRNA expression of interleukin-1 beta (IL-1β), -6, -8 and TNF-α receptors (TNFR) 1 and 2. In contrast, ANG increased the mRNA expression of IL-4 and -10. Protein levels of TANK-binding kinase 1 (TBK1) were reduced by ANG in HCFs treated with TNF-α. Moreover, ANG diminished the expression of IL-6 and -8 and monocyte chemotactic protein- (MCP-) 1. The protein expression of nuclear factor-κB (NF-κB) was downregulated by ANG treatment. These findings suggest that ANG suppressed the TNF-α-induced inflammatory response in HCFs through inhibition of TBK1-mediated NF-κB nuclear translocation. These novel results are likely to play a significant role in the selection of immune-mediated inflammatory therapeutic targets and may shed light on the pathogenesis of immune-mediated inflammatory diseases.

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Related in: MedlinePlus

Western blot analyses of NF-κB in the nucleus and cytoplasm. (a) NF-κB nuclear translocation induced by TNF-α was diminished after ANG treatment. ANG treatment alone did not affect NF-κB nuclear translocation. (b) Densitometric analysis of the relative ratio of NF-κB in cells treated with TNF-α alone to those treated with TNF-α+ANG. The experiments were performed in triplicate (*P < 0.05). (c) Immunocytochemistry of NF-κB in the nuclei and cytoplasm of HCFs. After treatment with ANG, the cells were fixed and then labeled with an anti-NF-κB antibody. Immunofluorescent images at higher magnification demonstrate attenuation of the expression of NF-κB in nucleus after treatment with ANG ((c) arrow) in cells treated with TNF-α (scale bar, 100 μm).
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fig5: Western blot analyses of NF-κB in the nucleus and cytoplasm. (a) NF-κB nuclear translocation induced by TNF-α was diminished after ANG treatment. ANG treatment alone did not affect NF-κB nuclear translocation. (b) Densitometric analysis of the relative ratio of NF-κB in cells treated with TNF-α alone to those treated with TNF-α+ANG. The experiments were performed in triplicate (*P < 0.05). (c) Immunocytochemistry of NF-κB in the nuclei and cytoplasm of HCFs. After treatment with ANG, the cells were fixed and then labeled with an anti-NF-κB antibody. Immunofluorescent images at higher magnification demonstrate attenuation of the expression of NF-κB in nucleus after treatment with ANG ((c) arrow) in cells treated with TNF-α (scale bar, 100 μm).

Mentions: HCFs were either cultured with TNF-α (20 ng/mL, 8 h) or treated with ANG (2 μg/mL, 0.5 h) to examine whether ANG inhibits nuclear translocation of NF-κB. The cells were subjected to immunofluorescent localization of NF-κB. Treatment with TNF-α induced the translocation of NF-κB from the cytosol to nucleus. However, the presence of ANG inhibited the translocation of NF-κB (Figure 5). ANG treatment alone did not influence the expression of NF-κB in nucleus.


Angiogenin reduces immune inflammation via inhibition of TANK-binding kinase 1 expression in human corneal fibroblast cells.

Lee SH, Kim KW, Min KM, Kim KW, Chang SI, Kim JC - Mediators Inflamm. (2014)

Western blot analyses of NF-κB in the nucleus and cytoplasm. (a) NF-κB nuclear translocation induced by TNF-α was diminished after ANG treatment. ANG treatment alone did not affect NF-κB nuclear translocation. (b) Densitometric analysis of the relative ratio of NF-κB in cells treated with TNF-α alone to those treated with TNF-α+ANG. The experiments were performed in triplicate (*P < 0.05). (c) Immunocytochemistry of NF-κB in the nuclei and cytoplasm of HCFs. After treatment with ANG, the cells were fixed and then labeled with an anti-NF-κB antibody. Immunofluorescent images at higher magnification demonstrate attenuation of the expression of NF-κB in nucleus after treatment with ANG ((c) arrow) in cells treated with TNF-α (scale bar, 100 μm).
© Copyright Policy - open-access
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC4016892&req=5

fig5: Western blot analyses of NF-κB in the nucleus and cytoplasm. (a) NF-κB nuclear translocation induced by TNF-α was diminished after ANG treatment. ANG treatment alone did not affect NF-κB nuclear translocation. (b) Densitometric analysis of the relative ratio of NF-κB in cells treated with TNF-α alone to those treated with TNF-α+ANG. The experiments were performed in triplicate (*P < 0.05). (c) Immunocytochemistry of NF-κB in the nuclei and cytoplasm of HCFs. After treatment with ANG, the cells were fixed and then labeled with an anti-NF-κB antibody. Immunofluorescent images at higher magnification demonstrate attenuation of the expression of NF-κB in nucleus after treatment with ANG ((c) arrow) in cells treated with TNF-α (scale bar, 100 μm).
Mentions: HCFs were either cultured with TNF-α (20 ng/mL, 8 h) or treated with ANG (2 μg/mL, 0.5 h) to examine whether ANG inhibits nuclear translocation of NF-κB. The cells were subjected to immunofluorescent localization of NF-κB. Treatment with TNF-α induced the translocation of NF-κB from the cytosol to nucleus. However, the presence of ANG inhibited the translocation of NF-κB (Figure 5). ANG treatment alone did not influence the expression of NF-κB in nucleus.

Bottom Line: Although ANG may be involved in the anti-inflammatory response, there is no evidence that it has direct anti-inflammatory effects.These findings suggest that ANG suppressed the TNF-α-induced inflammatory response in HCFs through inhibition of TBK1-mediated NF-κB nuclear translocation.These novel results are likely to play a significant role in the selection of immune-mediated inflammatory therapeutic targets and may shed light on the pathogenesis of immune-mediated inflammatory diseases.

View Article: PubMed Central - PubMed

Affiliation: Department of Ophthalmology, College of Medicine, Chung-Ang University Hospital, 224-1 Heukseok-dong, Dongjak-Gu, Seoul 156-755, Republic of Korea ; Department of Medicine, Graduate School, Chung-Ang University, Seoul, Republic of Korea.

ABSTRACT
Angiogenin (ANG) is reportedly multifunctional, with roles in angiogenesis and autoimmune diseases. This protein is involved in the innate immune system and has been implicated in several inflammatory diseases. Although ANG may be involved in the anti-inflammatory response, there is no evidence that it has direct anti-inflammatory effects. In this study we sought to determine whether ANG has an anti-inflammatory effect in human corneal fibroblasts (HCFs) exposed to media containing tumor necrosis factor-alpha (TNF-α). We found that ANG reduced the mRNA expression of interleukin-1 beta (IL-1β), -6, -8 and TNF-α receptors (TNFR) 1 and 2. In contrast, ANG increased the mRNA expression of IL-4 and -10. Protein levels of TANK-binding kinase 1 (TBK1) were reduced by ANG in HCFs treated with TNF-α. Moreover, ANG diminished the expression of IL-6 and -8 and monocyte chemotactic protein- (MCP-) 1. The protein expression of nuclear factor-κB (NF-κB) was downregulated by ANG treatment. These findings suggest that ANG suppressed the TNF-α-induced inflammatory response in HCFs through inhibition of TBK1-mediated NF-κB nuclear translocation. These novel results are likely to play a significant role in the selection of immune-mediated inflammatory therapeutic targets and may shed light on the pathogenesis of immune-mediated inflammatory diseases.

Show MeSH
Related in: MedlinePlus