p38MAPK信号通路在盐酸戊乙奎醚减轻内毒素诱导人脐静脉内皮细胞损伤中的作用

Role of p38 MAPK signal pathway in attenuation of iipopolysaccharide-induced human umbilical vein endo- thelial cell injury by penehycHdine hydrochloride

目的探讨p38丝裂原活化蛋白激酶（p38MAPK）信号通路在盐酸戊乙奎醚减轻内毒素诱导人脐静脉内皮细胞损伤中的作用。方法培养人脐静脉内皮细胞，以1×104／ml密度接种于96孔培养板（100μ／孔）或24孔培养板（3ml／孔），以1×106／ml密度接种于培养瓶（5ml／瓶），采用随机数字表法，将其随机分为4组（n=23）：正常对照组（C组）、脂多糖（LPS）组（L组）、盐酸戊乙奎醚组（P组）和盐酸戊乙奎醚＋LPS组（PL组），P组和PL组加入终浓度为2μg／ml的盐酸戊乙奎醚，L组和PL组加人终浓度为1μg／ml的LPS，PL组于加入盐酸戊乙奎醚后1h加入LPS。加入LPS后24h时，收集细胞，测定磷酸化p38MAPK（p-p38MAPK）和p38MAPK的表达水平，以二者比值反映p38MAPK激活程度，并测定细胞活力、NO含量和诱导型一氧化氮合酶（iNOS）表达。结果与C组比较，L组细胞活力降低，NO、p-p38MAPK和p38MAPK激活程度升高，iNOS表达上调（P〈0．01），P组上述指标差异无统计学意义（P〉O．05）；与L组比较，PL组细胞活力升高，NO、p-p38MAPK和p38MAPK激活程度降低，iNOS表达下调（P〈0．05或0．01）。结论p38MAPK信号通路参与了盐酸戊乙奎醚减轻内毒素诱导人脐静脉内皮细胞损伤的作用。

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/d/hole） or 24-well plate （3 ml/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 tzg/ml in groups L and PL or/and PHC 2 /lg/ml in groups P and PL. LPS was added at 1 h after PHC in group PL. The ceils 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 1〉38 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.