盐酸戊乙奎醚对内毒素刺激的肺微血管内皮细胞的作用及对β-抑制蛋白-2的影响

Effect of Penehyclidine Hydrochloride on Endothelial Injury and Expression of Beta-Arrestin-2 in LPS-Induced HPMVEC

目的:探讨在内毒素(LPS)作用下盐酸戊乙奎醚(PHCD)对肺微血管内皮细胞的作用及β-抑制蛋白-2(β-arrestin-2)表达的影响。方法:培养人肺微血管内皮细胞,用随机数字表法将其分为4组:正常对照组(C组)、LPS诱导组(L组)、PHCD+LPS组(PL组)及PHCD对照组(P组)。P组和PL组加入终浓度为2μg/ml的PHCD,L组及PL组加入终浓度为0.1μg/ml的LPS,PL组于加入PHCD后60min再加入LPS,C组加入同等体积的培养基。加入LPS 60min后,测细胞LDH水平、VCAM-1、VE-cadherin表达及β-arrestin-2的表达。结果:与C组比较,L组细胞LDH水平、VCAM-1表达升高,VE-cadherin和β-arrestin-2表达降低;与L组比较,PL组细胞LDH水平、VCAM-1表达降低,而VE-cadherin和β-arrestin-2表达增加。结论:盐酸戊乙奎醚预处理,可以通过上调β-arrestin-2的表达,减轻LPS引起的肺微血管内皮细胞损伤。

Objective：To investigate the effect of penehyclidine hydrochloride（PHCD）on endothelial injury and expression ofβ-arrestin-2 in lipopolysaccharide（LPS）-induced human pulmonary microvascular endothelial cell（HPMVEC）.Methods：HPMVEC were cultured and seeded in 6-well plates（2ml/hole）with the density of 2×105/ml or in culture flasks（4ml/flask）with the density of 1×107/ml,then,cells were randomly divided into 4groups：group control（group C）,group LPS（group L）,group PHCD＋LPS（group PL）and group PHCD（group P）.The cells were treated with 2μg/ml PHCD in group P.In group PL,cells were treated with 2μg/ml PHCD for 60 min and then stimulated with 0.1μg/ml LPS.Cells in group C were cultured with the same amount of medium.Then,cells were collected at 60 min after LPS treatment to measure thelevel of LDH,and the expression of VCAM-1,VE-cadherin and β-arrestin-2.Results：Compared with group C,the level of LDH and expression of VCAM-1 were increased,and the expression of VE-cadherin andβ-arrestin-2 were decreased in group L.Compared with group L,the level of LDH and expression of VCAM-1 were decreased,and VE-cadherin andβ-arrestin-2 expression were increased in group PL.Conclusion：These findings suggest that PHCD can alleviate the LPS-induced pulmonary microvascular endothelial injury by increasing the expression ofβ-arrestin-2.