Role of p38 MAPK signal pathway in attenuation of lipopolysaccharide-induced human umbilical vein endothelial cell injury by penehyclidine hydrochloride

Objective To investigate the role of p38 mitogen-activated protein kinase (p38 MAPK) in attenuation of lipopolysaccharide (LPS)-induced human umbilical vein endothelial cell injury by penehyclidine hydrochloride (PHC).Methods Human umbilical vein endothelial cells were provided by Medical Research Center,Wuhan University,cultured and seeded in 96-well plate (100 μl/hole) or 24-well plate (3 nl/hole) with density of 1 × 104/ml or in culture flasks (5 ml/flask) with density of 1 × 106/ml.The cells were randomly divided into 4 groups ( n =23 each):group control (group C) ; group LPS; group PHC (group P) and group PHC + LPS (group PL).The cells were exposed to LPS 1 μg/ml in groups L and PL or/and PHC 2 μg/ml in groups P and PL.LPS was added at 1 h after PHC in group PL.The cells were collected at 24 h exposure to LPS for determination of the expression of phosphorylated p38 MAPK (p-p38 MAPK) and p38 MAPK.The ratio between p-p38 MAPK and p38 MAPK was calculated.Cell viability,NO content and inducible nitric oxide synthase (iNOS) expression were also determined.Results LPS significantly decreased cell viability,increased NO content,iNOS expression,p-p38 MAPK and p-p38 MAPK/p38 MAPK ratio in group L as compared with group C.In group PL pretreatment with PHC significantly attenuated LPS-induced cell injury.Conclusion p38 MAPK pathway is involved in attenuation of LPS-induced endothelial cell injury by PHC. Key words: p38 Mitogen-activated protein kinases; Cholinergic antagonists; Endotoxemia; Endothelium,vascular