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World Journal of Emergency Medicine ›› 2017, Vol. 8 ›› Issue (4): 292-296.doi: 10.5847/wjem.j.1920-8642.2017.04.008

• Original Articles • Previous Articles     Next Articles

The effects of microRNA-34a regulating Notch-1/NF-κB signaling pathway on lipopolysaccharide-induced human umbilical vein endothelial cells

Yun Ge, Man Huang, Yue-feng Ma()   

  1. Department of General Intensive Care Unit, The Second Affiliated Hospital, Zhejiang University School of Medicine, Hangzhou 310052, China
  • Received:2017-02-27 Accepted:2017-07-20 Online:2017-12-15 Published:2017-12-15
  • Contact: Yue-feng Ma E-mail:2193017@zju.edu.cn

Abstract:

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 μg/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.

Key words: MicroRNA-34a, Notch-1, NF-κB, Lentivirus, Human umbilical vein endothelial cells