The Effects of Interleukin-17 (IL-17)-Related Inflammatory Cytokines and A20 Regulatory Proteins on Astrocytes in Spinal Cord Cultured In Vitro

Abstract

Background/Aims: This study focused on investigating the expression of several inflammatory cytokines and chemokines, including regulatory proteins in the astrocytes of mice stimulated with IL-17. Materials and Methods: The cultured astrocytes from the spinal cords of mice were stimulated with IL-17. The expression of interleukin-6 (IL-6), tumor necrosis factor (TNF), monocyte chemotactic protein-1/5 (MCP-1/5) and macrophage inflammatory protein-2 (MIP-2) stimulated with IL-17 (50 ng/ml) at different time points (3 h, 6 h, 12 h, 24 h and 48 h) were determined using real-time PCR and ELISA. The expressions of A20 (tumor necrosis factor a inducible protein 3, TNFAIP3) and NF-κB were examined using real-time PCR and western blotting. Results: Compared with the control group, the mRNA expression levels of IL-6, TNF, MCP-1/5 and MIP-2 increased significantly at 6 h after IL-17 stimulation, while the protein expression levels also increased significantly and peaked at 12 h. The mRNA expression level of NF-κB increased and peaked at 6 h before gradually declining, while the expression of A20 decreased. The protein expression level of NF-κB increased and peaked at 12 h, while the expression A20 had an opposite response. Conclusion: The study showed that NF-κB may have an effect on the cytokines secreted by astrocytes after IL-17 stimulation. Moreover, both A20 and NF-κB could regulate the expression and secretion of inflammatory mediators.

Keywords

• Interleukin-17
• Astrocyte
• Inflammatory cytokines
• Chemokines
• A20