Figure 2

ENS activation in rat primary cultures inhibits the LPS-induced increase in TNF-α transcript and protein levels. (A) Evaluation of the impact of electrical field stimulation (EFS) on TNF-α secretion was performed by ELISA in rENSpc treated (+) or not (−) with LPS (0.1 μg/ml) (twelve independent experiments). (B) TNF-α mRNA expression was measured in rENSpc by qPCR (twelve independent experiments). (C) The effect of ATP (100 μM) and involvement of P2X7 receptor on TNF-α secretion were measured by ELISA in ENS cultures pretreated or not with BzATP (P2X7 agonist; 100 μM) or A439079 (P2X7 antagonist; 30 μM) (five independent experiments). 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 or antagonist versus LPS alone; §P <0.05 LPS with ATP compared with LPS with ATP in presence of A438079). ATP, adenosine-5'-triphosphate; BzATP, 2’(3’)-O-(4-benzoylbenzoyl) adenosine-5'-triphosphate triethylammonium salt; CT, control; EFS, electrical field stimulation; LPS, lipopolysaccharide; mRNA, messenger ribonucleic acid; rENSpc, rat enteric nervous system primary culture; SEM, standard error of the mean; TNF-α, tumor necrosis factor alpha.