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BACKGROUND: Notch-1/NF-κB signaling plays a key role in the cecal ligation and puncture(CLP)-induced sepsis. This study aims to investigate the intervention effects of microRNA-34a(miR-34a) lentivirus regulating Notch-1/NF-κB signaling pathway on lipopolysaccharide(LPS)-induced human umbilical vein endothelial cells(HUVEC).METHODS: HUVEC were divided into four groups as the following: they were infected with negative control lentivirus(NC group) or miR-34a lentivirus(OE group); LPS(1 g/mL) was added on the third day on the basis of NC group and OE group for 24 hours(NC+LPS group or OE+LPS group). The levels of TNF-α, IL-1β, IL-6, and IL-10 in the cell supernatants, and the mRNA and protein expression of Notch-1 and NF-κB in the HUVEC were evaluated.RESULTS: After 24 hours, the levels of TNF-α, IL-1β, IL-6 in the cell supernatants and the protein expression of NF-κB from NC+LPS group were significantly higher than those of NC group, but IL-10 level and the protein expression of Notch-1 in NC+LPS group were the opposite. After intervention of miR-34a lentivirus, the cell supernatants TNF-α and the protein expression of NF-κB in OE+LPS group after 24 hours markedly decreased compared to NC+LPS group. While the cell supernatants IL-1β and IL-6 and the mRNA expression of NF-κB slightly decreased in OE+LPS group, IL-10 and the mRNA and protein expression of Notch-1 were the opposite.CONCLUSION: miR-34a regulating Notch-1/NF-κB signaling pathway can reduce the HUVEC damage caused by LPS stimulation.
BACKGROUND: Notch-1 / NF-κB signaling plays a key role in the cecal ligation and puncture (CLP) -induced sepsis. This study aims to investigate the intervention effects of microRNA-34a (miR-34a) lentivirus regulating Notch-1 / NF-κB signaling pathway on lipopolysaccharide (LPS) -induced human umbilical vein endothelial cells (HUVEC). METHODS: HUVEC were divided into four groups as the following: they were infected with negative control lentivirus (NC group) or miR-34a lentivirus OE group); LPS (1 g / mL) was added on the third day on the basis of NC group and OE group for 24 hours (NC + LPS group or OE + LPS group). The levels of TNF- 1β, IL-6, and IL-10 in the cell supernatants, and the mRNA and protein expression of Notch-1 and NF-κB in the HUVEC were 35. RESULTS: After 24 hours, the levels of TNF- 1β, IL-6 in the cell supernatants and the protein expression of NF-κB from NC + LPS group were significantly higher than those of NC group, but IL-10 level and the protein expression of After intervention of miR-34a lentivirus, the cell supernatants TNF-α and the protein expression of NF-κB in OE + LPS group after 12 hours markedly decreased compared to NC + LPS group . While the cell supernatants IL-1β and IL-6 and the mRNA expression of NF-κB slightly decreased in OE + LPS group, IL-10 and the mRNA and protein expression of Notch-1 were the opposite. regulating Notch-1 / NF-κB signaling pathway can reduce the HUVEC damage caused by LPS stimulation.