Effects of thiol on NF-?B activity in and release of TNF-? from endotoxin-activated cultured rat kupffer cells / 中华麻醉学杂志

ABSTRACT

Objective To determine whether thiol can affect the activity of nuclear factor kappa B (NF-?B) in and release of TNF-?from lipopolysaccharide (LPS)-activated kupffer cells (KCs) and ascertain whether these effects are mediated through glutathione (GSH) .Methods KCs were isolated from livers of healthy male adult SD rats weighing 250-300g and cultured for 12h. The cultured KCs were exposed to LPS 10 ng?ml-1. The effects of different concentrations of glutathione monoethylester (GSHEE) and N-acetylcysteine (NAC) on NF-?B activity in and release of TNF-afrom KCs were determined. The effect of pretreatment with GSH synthesis inhibitor - BSO on the inhibitory effect of NAC on inflammation was investigated. NF?B activity was determined by electrophoretic mobility shift assay (EMSA) and intracellular GSH content and TNF-?level in supernatant by HPLC. Results Intracellular GSH content remained unchanged after exposure of KCs to LPS. Both GSHEE and NAC increased intracellular GSH level and inhibited NF-?B activity and release of TNF-?. BSO blocked the increase in GSH induced by NAC but did not affect the inhibitory effect of NAC on TNF?release. Conclusions LPS does not affect GSH level in KCs. NAC can inhibit NF-?B activity in and TNF-?release from KCs and increase intracellular GSH level but the inhibitory effect is not mediated through GSH.