Effect and mechanism of rat macrophage α7-acetylcholine receptor mediated anti-inflammatory ;pathway on endotoxin-induced inflammation reaction / 中华临床感染病杂志

ABSTRACT

Objective To investigate the effect of rat macrophage α7-acetylcholine receptor (α7-AChR )-mediated cholinergic anti-inflammatory pathway on endotoxin-induced inflammation reaction . Methods Density gradient centrifugation method was used to isolate rat primary macrophages and flow cytometry was used to identify the cell purity .α7-AChR in macrophages was detected by fluorescence confocal microscopy and Western blot .After 1ipopolysaccharides ( LPS) was added to the culture media of primary culture of macrophages , the concentration of tumor necrosis factor ( TNF)-αin the supernatant was determined by enzyme linked immunosorbent assay (ELISA).The expression of p-NF-κB protein in primary cultured macrophages was detected by Western blot .ANOVA was used to analyze TNF-αlevels after adding different concentrations of nicotine .Results α7-AChR was observed by fluorescence confocal microscope in primary macrophages .Nicotine could significantly reduce the concentration of TNF-αin culture supernatants of macrophages after LPS stimulation .When the concentrations of nicotine were 0, 1, 10, 100μmol/L, the concentrations of TNF-αwere (2 123 ±86), (1 486 ±80), (1 316 ±83) and (1 090 ±77)pg/mL, respectively (t=16.33, 20.18 and 26.83, P<0.05).The level of p-NF-κB in macrophages was also reduced when nicotine added .Conclusion Activation of macrophage α7-AChR can inhibit the endotoxin-induced release of inflammatory factor TNF-α, which may be through NF-κB signal pathway .