Role of Erbin in muramyl dipeptide-induced inflammatory responses in macrophages of mice / 中华麻醉学杂志

ABSTRACT

Objective:To evaluate the role of ErbB2-interacting protein (Erbin) in muramyl dipeptide (MDP)-induced inflammatory responses in the macrophages of mice.Methods:Erbin gene knockout RAW264.7 cell line (Erbin -/ -RAW264.7) was constructed by CRISPR/CAS9 gene-editing technology.RAW264.7 cells were cultured in vitro.Each type of cells was divided into 2 groups ( n=16 each)by a random number table method: RAW264.7 group, RAW264.7 plus MDP group, erbin -/ -RAW264.7 group, and erbin -/ -RAW264.7 plus MDP group.In each MDP group, cells were incubated with 10 μg/ml MDP for 6 h, then immunofluorescence was used to determine the expression of nuclear factor kappa B (NF-κB) p65, and the concentrations of tumor necrosis factor-alpha(TNF-α)and interleukin-6(IL-6)in the culture medium were determined by enzyme-linked immunosorbent assay. Results:Compared with RAW264.7 group, the concentrations of TNF-α and IL-6 in the culture medium were significantly increased( P<0.05), NF-κB p65 moved to the nucleus, and the red fluorescence area was increased in RAW264.7+ MDP group.Compared with RAW264.7+ MDP group and Erbin -/- RAW264.7 group, the concentrations of TNF-α and IL-6 in the culture medium were significantly increased ( P<0.05), NF-κB p65 moved more markedly to the nucleus, and the red fluorescence area was increased in Erbin -/-RAW264.7+ MDP group. Conclusion:Erbin inhibits MDP-induced inflammatory responses in macrophages through inhibiting the activity of NF-κB p65 in mice.