脐血与成人外周血血浆对脂多糖诱导的血管内皮细胞TLR4/NF-κB表达的影响

The impact of umbilical cord plasma versus that of adult peripheral plasma on lipopolysaccharide-induced TLR4/NF-κB expression in the vascular endothelial cells

目的:探讨脐血血浆与成人外周血血浆对脂多糖(LPS)诱导的血管内皮细胞TLR4/NF-κB表达的影响。方法:采集剖宫产分娩的足月健康孕妇脐血和健康成人外周血各20份,制备血浆。将体外培养的人脐静脉内皮细胞ECV304分为两组,(1)脐血组,加入1μg/mL的LPS和10%的脐血血浆;(2)成人外周血组,加入1μg/mL的LPS和10%的成人外周血血浆。逆转录-聚合酶链反应检测TLR4 mRNA的表达,免疫印迹法检测TLR4、IκBα蛋白的表达。结果:脐血组TLR4 mRNA及TLR4蛋白的表达量显著低于成人外周血组(P<0.01),脐血组IκBα的蛋白表达量显著高于成人外周血组(P<0.01)。结论:脐血血浆抑制LPS诱导的内皮细胞TLR4/NF-κB活化,从而抑制其炎症反应。

Objective To compare the impact of umbilical cord plasma with that of adult peripheral plasma on lipopolysaccharide-induced TLR4/NF-κB expression in the vascular endothelial cells. Methods Umbilical cord blood from 20 full-term infants and peripheral venous blood from 20 healthy adult volunteers were collected to generate plasma. Human umbilical vascular endothelial cells （ECV304） were cultured in vitro with 1 μg/mL LPS plus either 10% umbilical cord plasma （cord plasma group） or 10% peripheral plasma （peripheral plasma group）. Toll-like receptor 4 （TLR4） mRNA expression was detected by RT-PCR, so were expressions of TLR4 and IKBα protein by Western blot. Results The expressions of TLR4 mRNA and TLR4 protein were significantly lower but IκBα protein expression was higher in the cord plasma group, as compared with the peripheral plasma group （P 〈 0.01 for all comparisons）. Conclusions Umbilical cord plasma can inhibit the LPS-induced TLR4/NF-κB activation, resulting in the suppression of the inflammatory reaction.