长托宁对脂多糖引起的人脐静脉内皮细胞损伤的保护作用及其机制  

Protective effects of penehyclidine hydrochloride on lipopolysaccharide-induced human umbilical vein endothelial cells injury and its mechanism

作　　者：罗金龙[1] 谢艾妮[2] 刘环芹[1] 杨鹏[1] 李树生[1] 杨光田[1] 

机构地区：[1]华中科技大学同济医学院附属同济医院急诊科,武汉430030 [2]华中科技大学同济医学院附属协和医院心血管外科

出　　处：《中华实验外科杂志》2012年第7期1299-1302,共4页Chinese Journal of Experimental Surgery

摘　　要：目的观察长托宁（PHC）对脂多糖（LPS）引起的人脐静脉内皮细胞（HUVEC）损伤的保护作用，并探讨其机制。方法将体外培养HUVEC分为5组：空白对照组、LPS组、LPS／PHC（10μg/L）组、LPS／PHC（25μg/L）组、LPS／PHC（50μg/L）组。噻唑蓝（MTT）比色法测定内皮细胞的存活率；化学比色法测定乳酸脱氢酶（LDH）浓度；硝酸还原酶法测定一氧化氮（NO）浓度；定量逆转录-聚合酶链反应（RT—PCR）检测诱导型一氧化氮合酶（iNOS）mRNA表达；Western blot法检测iNOS蛋白质含量；酶联免疫吸附试验（ELISA）测定核因子-κB（NF—κB）和信号转导和转录激活因子3（STAT3）活性。结果与空白对照组比较，LPS组细胞存活率[（60．31±8．76）％比（100．00±0．00）％]明显下降、LDH含量[（326±52）μmol／L比（125±25）μmol／L]和NO浓度[（55．49±10．16）μmol／L比（12．13±11．02）μmol／L]明显增加（P〈0．01）；而PHC可以逆转上述效应（P〈0．05）。同时PHC可以降低LPS导致的内皮细胞iNOS转录和蛋白质表达水平，下调NF—κB和STAT3的表达（P〈0．05）。结论长托宁可以减轻LPS对内皮细胞的损伤，其作用可能是通过抑制NF—κB和STAT3信号系缔．减少NO生成宴王Ⅲ的．Objective To investigate the protective effects of penehyclidine bydrochloride （PHC） on lipopolysaceharide （LPS）-induced human umbilical vein endothelial cells （HUVECs） injm7 and the underlying mechanism. Methods Cultured HUVECs were divided into five groups： control group, LPS group, LPS/PHC （10 μg/L） group, LPS/PHC （25 μg/L） group, LPS/PHC （50 μg/L） group. Cell via- bility was measured by methyl thiazol tetrazolium （MTT） colorimetric method. The concentrations of lactate dehydrogenase （LDH） and nitric oxide （NO） were measured respectively by using chemical colorimetric method and nitrate reduction method. The expression of inducible nitric oxide synthase （iNOS） mRNA and protein was detected by using reverse transcription-polymerase chain reaction （RT-PCR） and Western blot- ting respectively. The activity of nuclear factor-κB （NF-κB） and signal transducer and activators of tran- scription 3 （STAT3） were measured by enzyme linked immunosorbent assay （ELISA）. Results As com- pared with control group, cell viability [ （60. 31 ± 8.76）% vs 100. 00% ] was decreased and the concen- trations of LDH [ （326 ± 52） μmol/L vs （ 125 ± 25 ） μmol/L ] and NO [ （ 55.49 ± 10. 16 ） μmol/L vs （12. 13 ± 11.02） μmol/L] were increased in LPS group （P 〈 0.01 ） ; PHC could reverse these effects oil HUVECs （P 〈0. 05）. At the same time, PHC reduced the expression of iNOS mRNA and protein, and the activity of NF-κB and STAT3 （P 〈 0. 05 ）. Conclusion PHC can protect the endothelial function by reducing the concentration of NO. Inhibiting the NF-κB and STAT3 signal pathway may be the underlying mechanism.

关 键 词：长托宁 脂多糖 人脐静脉内皮细胞 诱导型一氧化氮合酶 一氧化氮 

分 类 号：R285.5[医药卫生—中药学]