[Sophoridine suppresses inflammatory cytokine secretion by lipopolysaccharide-induced RAW264.7 cells and its mechanism].
摘要
OBJECTIVE: To observe the effects of sophoridine on lipopolysaccharide (LPS)-induced secretion of tumor necrosis factor α (TNF-α) and interleukin 1β (IL-1β) as well as the expressions of Toll-like receptor 4 (TLR4) and c-Jun in RAW264.7 cells and explore the molecular mechanism of anti-LPS of sophoridine. METHODS: RAW264.7 cells were cultured and divided into four groups: macrophage control group (using serum-free DMEM to incubate cells), sophoridine control group (using 31.25 mg/L sophoridine-added DMEM to incubate cells), LPS group and sophoridine intervention group (using 100 μg/L LPS DMEM to incubate cells for 60 minutes, then throwing away LPS and adding serum-free DMEM or 31.25 mg/L sophoridine DMEM to incubate cells). Cells and culture medium were collected respectively at 5, 30, 60 and 120 minutes after the above treatment. The expression levels of TLR4 and c-Jun mRNA were determined by reverse transcription PCR (RT-PCR), and the expression of c-Jun protein in RAW264.7 cells was measured by immunocytochemistry and Western blotting; The levels of TNF-α and IL-1β in cell culture medium were analyzed by ELISA. RESULTS: Compared with macrophage control group, sophoridine control group had no statistical difference in each index. Compared with macrophage control group, the expressions of TLR4 mRNA, c-Jun mRNA and protein as well as the secretion of TNF-α and IL-1β significantly increased at each time point in LPS group, and maintained the level to 120 minutes. Sophoridine suppressed the expressions of TLR4 mRNA, c-Jun mRNA and protein, and reduced the secretion of TNF-α and IL-1β in LPS-stimulated RAW264.7 cells in sophoridine intervention group. CONCLUSION: Sophoridine down-regulated the secretion of TNF-α and IL-1β in LPS-induced RAW264.7 cells via inhibiting the expressions of TLR4 and c-Jun.