Effect of PrP105-132 on the secretion of interleukin-6 and interleukin-8 from microglial cells in vitro

Abstract

In the present study, the effect of prion protein (PrP) on the secretion of interleukin-6 (IL-6) and IL-8 from microglial cells in vitro and its possible underlying pathway were investigating by establishing a cell model for prion disease. Rat neuroglial cells were cultured in vitro, and were treated with 80 μM PrP peptides 105-132 (PrP105-132) only, PrP+MG132 or PrP+cyclosporin A (CsA). After 48 h, the IL-6 and IL-8 levels in the supernatant fluid of the treated cells were detected using enzyme-linked immunosorbent assay. In addition, the expression levels of nuclear factor-κB (NF-κB) and nuclear factor of activated T cells (NFAT) were evaluated using reverse transcription-polymerase chain reaction. The results indicated that the microglial cells were activated by treatment with PrP peptides. Cell bodies were augmented and appeared to have round, rod and amoeba-like shapes. In addition, the protuberances were shortened and eventually disappeared. Furthermore, the mRNA expression levels of NF-κB and NFAT in microglial cells increased, as well as the IL-6 and IL-8 levels in the supernatant fluid after treatment with PrP. However, the mRNA expression levels of NF-κB, and the IL-6 and IL-8 levels decreased after these cells were treated with MG132, a specific inhibitor of NF-κB. The mRNA expression of NFAT decreased after these cells were treated with CsA, a specific inhibitor of NFAT; however, the IL-6 level decreased, while no significant difference was observed in the IL-8 level. In conclusion, PrP-treated microglial cells secreted IL-6 and IL-8, and the secretion of IL-6 was associated with the activation of NF-κB and NFAT pathways. In addition, the secretion of IL-8 was mainly dependent on the NF-κB pathway.