Study on NF-κB Activation Pathway in rVvhA-effected THP-1 Cells

Abstract: This study designed to investigate how recombinant Vibrio vulnificus hemolysin (rVvhA) actives NF-κB signaling pathway in the PMA-stimulated THP-1 cell line. The cytotoxic effect of rVvhA on the growth of THP-1 cells was identified by CCK-8. Cell morphology was observed by inverted microscope. NF-κB p65 nuclear transfer was observed by laser scanning confocal microscope. NF-κB p65 expression in rVvhA effected THP-1 cells was detected by flow cytometry. NF-κB p65 expression in rVvhA effected THP-1 cell nucleus was determined by Western blot analysis. TNF-α and IL-6 expression in rVvhA effected THP-1 cells was quantified by ELISA. The viability of THP-1 cells exposed to rVvhA was inhibited in time-dose dependent mannar and the cell morphology changed obviously observed by inverted microscope. The levels of NF-κB p65 transferred into nuclear reached its tiptop after treated by 0.4 HU/mL rVvhA for 6 h. Total content of NF-κB p65 in cells peaked at 2 h treated by 0.6 HU/mL rVvhA. NF-κB p65 in cell nucleus was the most after 0.6 HU/mL rVvhA treated for 4 h. rVvhA effected THP-1 cells can raise the expressions of TNF-α and IL-6. NF-κB inhibitors can restrain the expression of IL-6. rVvhA had cytotoxic effect on THP-1 cells. It can activate NF-κB signaling pathway in THP-1 cells and increase the expressions of TNF-α and IL-6， meanwhile， NF-κB inhibitors can decrease the expression level of IL-6.

CSTR: 32200.14.cjcb.2012.02.0007