LOC339524 gene mediates lipopolysaccharide-induced expression of inflammatory cytokines in mouse microglia bv2 cell line

Abstract

Objective To study the role of LOC339524 gene in mediating the expression of inflammatory cytokines in mouse microglia bv2 cell line induced by lipopolysaccharide (LPS). Methods Mouse microglia bv2 cells were treated with 100 ng/mL LPS for 3, 6, 9 or 12 h to determine the optimal time for observing the inflammatory response. The cells were then treated with 10, 100, 1 000, and 10 000 ng/mL LPS or with 100 ng/mL LPS for 3, 6, 9, and 12 h, and the changes in the expression of LOC339524 protein was investigated. The changes in the expression of tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6) in response to LPS challenge were detected in bv2 cells with siRNA-mediated LOC339524 gene silencing or lentivirus-mediated LOC339524 overexpression using qPCR and Western blotting. Results Compared with blank control, LPS treatment increased TNF-α, IL-6 mRNA and protein expression at different times (P < 0.05), and the expression was consistent at 12 hours. 12 hours were selected for subsequent inflammatory reaction studies; The expression of LOC339524 protein was increased viadifferent concentrations of LPS treatment(P < 0.05) when compared with 0 ng/mL, and that increase was in a time-dependent manner at 100 ng/mL LPS concentration (P < 0.05). After 12 hours of LPS treatment, the expression of TNF-α and IL-6 mRNA and protein was significantly up-regulated in the siRNA group compared with the unrelated fragment group (P < 0.05). Conversely the expression of TNF-α and IL-6 mRNA and protein in the over-expression group was significantly lower than that in the normal group (P < 0.05). Conclusion LOC339524 is an inflammatory response factor that promotes the inflammatory response of bv2 cells to LPS.

Keywords

• ] lipopolysaccharide
• bv2 cells
• loc339524 gene
• tumor necrosis factor
• interleukin-6