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Recombinant Legionella pneumophila flagella protein A (rflaA) induces the secretion of IL-6 and IL-1β in RAW264.7 cells in vitro
Ist Teil von
Xi bao yu fen zi mian yi xue za zhi, 2017-05, Vol.33 (5), p.601
Ort / Verlag
China
Erscheinungsjahr
2017
Link zum Volltext
Quelle
MEDLINE
Beschreibungen/Notizen
Objective To investigate the effect of recombinant Legionella pneumophila flagella protein A (rflaA) on the secretion of interleukin-6 (IL-6) and interleukin-1β (IL-1β) by RAW264.7 macrophage and the possible mechanism. Methods RAW264.7 cells were treated with 0.000, 0.125, 0.250, 0.500, 1.000, 2.000, 4.000 and 8.000 μg/mL rflaA to determine the EC
of rflaA using CCK-8 assay. Secretion of IL-6 and IL-1β were measured by ELISA at 24, 36 and 48 hours after treatment of the cells with 0.04, 0.08 and 0.16 μg/mL rflaA. At 6, 12, 24, 36 and 48 hours after treatment of the cells with 0.04, 0.08 and 0.16 μg/mL rflaA, the expressions of IL-6, IL-1β, NOD-like receptor protein 3 (NLRP3) and caspase-1 mRNAs were detected by quantitative real-time PCR, and the expressions of NLRP3 and caspase-1 proteins were tested by Western blotting. Results RflaA enhanced the expressions of IL-6 and IL-1β, and the higher concentration of rflaA was more potential. The expressions of IL-6 and IL-1β reached peak when the cells were treate