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Kyung Sup Lim, Seung Hoon Lee, Kyoung Woo Kim, Jae Chan Kim; Angiogenin reduces immune-inflammation via inhibition of TANK-binding kinase 1 (TBK1) expression in human corneal fibroblast cells(HCFs). Invest. Ophthalmol. Vis. Sci. 2014;55(13):3226.
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To investigate the anti-inflammatory activity of ANG in TNF-α-induced inflamed HCFs that contribute to the regulation of corneal stromal inflammation through the regulation of production of cytokines and chemokines. Thus, we attempted to clarify the molecular activity of ANG for inhibiting TNF-α mediated transduction that involves the production of TBK1 and nuclear translocation of NF-κB.
Cultured HCFs were treated with TNF-α. In the time of treatment, the cells were treated with or without ANG (2μg/ml) for 30 minutes. We analysed TNF-α receptor(TNFR)1,2, interleukin-1beta (IL-1β), -4, -6, -8, -10, monocyte chemotactic protein (MCP)-1, TBK1, nuclear factor-κB (NF-κB) by RT PCR, Immunodot blot, Western blot analysis, Immunocytochemistry.
ANG was found to reduce the mRNA expression of IL-1β, -6, -8, TNFR 1 and 2 increased by TNF-α. Whereas, ANG increased the mRNA expression of IL-4 and -10. We found that ANG decreased the protein levels of TANK-binding kinase 1 (TBK1) in HCFs inflamed by TNF-α. Moreover, ANG diminished the expression of IL-6, -8 and MCP-1 in media of HCFs inflamed by TNF-α. We also detected that the protein expression of NF-κB in nuclei increased by TNF-α was down-regulated by ANG treatment.
ANG suppresses inflammatory response in HCFs inflammed by TNF-α through an inhibition of TBK1 mediated NF-κB nuclear translocation. These newly identified results are likely to have a significant role in selection of immune-inflammatory therapeutic target and may therefore shed refreshing light on the development of immune-inflammatory disease.
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