Role of cannabinoid receptor 2 in macrophage pyroptosis induced by lipopolysaccharide in mice / 中华麻醉学杂志

ABSTRACT

Objective:To evaluate the role of cannabinoid receptor 2 (CB2R) in macrophage pyroptosis induced by lipopolysaccharide (LPS) in mice.Methods:Macrophage line RAW264.7 cells of mice were routinely cultured and divided into 3 groups ( n=18 each) using a random number table method: control group (group C), LPS group and LPS plus CB2R agonist HU308 group (group HU308). Group C received no mediation, LPS at the final concentration of 1 μg/ml was added in the other groups.After incubation for 15 min, HU308 with the final concentration of 10 μmol/L was added in group LPS+ HU308.All groups were then incubated for 6 h. The expression of nucleotide-binding oligomerization domain-like receptor containing pyrin domain 3 (NLRP3), caspase-1, caspase-11 and gasdermin D (GSDMD) mRNA was measured by real-time polymerase chain reaction, the expression of NLRP3, caspase-1, caspase-11, GSDMD and C-terminal domain of human GSDMD (GSDMD-C) was detected by Western blot, and the concentrations of interleukin-18 (IL-18) and IL-1β in the supernatant were determined by enzyme-linked immunosorbent assay.GSDMD-C/GSDMD ratio was calculated. Results:Compared with group C, the expression of NLRP3, caspase-1, caspase-11, GSDMD and GSDMD-C was significantly up-regulated, GSDMD-C/GSDMD ratio was increased, and the concentrations of IL-18 and IL-1β in the supernatant were increased in group LPS ( P<0.05). Compared with group LPS, the expression of NLRP3, caspase-1, caspase-11, GSDMD and GSDMD-C was significantly down-regulated, GSDMD-C/GSDMD ratio was decreased, and the concentrations of IL-18 and IL-1β were decreased in group HU308 ( P<0.05). Conclusion:CB2R is involved in macrophage pyroptosis induced by LPS in mice.