Figure 4

ENS activation in human LMMP inhibits the LPS-induced TNF-α production. (A) The impact of electrical field stimulation (EFS) on TNF-α secretion by the human LMMP treated (+) or not (−) with LPS (0.1 μg/ml) was performed by ELISA (four independent experiments each performed in triplicate; CT means control). (B) The involvement of P2X7 receptor on TNF-α secretion were measured by ELISA in human LMMP pretreated, or not, with BzATP (P2X7 agonist; 100 μM) before LPS stimulation (three independent experiments each performed in triplicate; CT means control). (C) ERK- and AMPK-dependent pathway contribution to TNFα production induced by LPS were assessed using MEK1/2 inhibitor (U0126; 10 μM) and AMPK inhibitor (C compound, CC; 10 μM) co-treatment on human LMMP treated, or not, with LPS for seven hours (three independent experiments each performed in triplicate; CT means control). Values represent the mean ± SEM (Mann-Whitney U test; *P <0.05 as compared with control without LPS; #P <0.05 LPS with EFS, agonist versus LPS alone). AMPK, 5’-AMP-activated protein kinase; BzATP, 2’(3’)-O-(4-benzoylbenzoyl) adenosine-5'-triphosphate triethylammonium salt; CT, control; CC, C compound; CT, control; EFS, electrical field stimulation; ERK, extracellular signal-regulated kinase; LMMP, longitudinal muscle/myenteric plexus; LPS, lipopolysaccharide; MEK1/2, mitogen-activated protein kinase kinase 1/2; SEM, standard error of the mean; TNF-α, tumor necrosis factor alpha.