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目的观察五没食子酰葡萄糖(PGG)对内毒素/脂多糖(LPS)的体外拮抗作用。方法应用生物传感技术测定PGG与脂质A(lipidA)的亲和力,动态浊度法测定PGG对LPS的中和作用。分离培养人外周血单核细胞(PBMC),观察不同浓度的PGG对LPS刺激下人PBMC释放肿瘤坏死因子(TNF)α、白细胞介素(IL)6的影响。结果不同浓度的PGG与脂质A发生结合反应,其反应的速率随浓度的增高而增快,二者的解离常数(Kd)值为5.2×10-7mol/L。PGG对LPS具有较强的中和作用。PGG在20mg/L以上的浓度时能够显著抑制LPS刺激下TNF-α和IL-6的释放,此时其TNF-α和IL-6水平分别从(1788±171)、(1966±232)ng/L降至(958±234)、(1351±99)ng/L(P<0.05),该作用具有一定的剂量效应关系。结论PGG可通过对LPS的结合及中和作用,在体外发挥拮抗LPS的作用。
Objective To observe the in vitro antagonism of pentaglucose (PGG) on endotoxin/lipopolysaccharide (LPS). Methods Biosensor technology was used to determine the affinity of PPG and lipidA, and dynamic nephelometry was used to determine the neutralizing effect of PPG on LPS. Human peripheral blood mononuclear cells (PBMCs) were isolated and cultured, and the effects of different concentrations of PPG on release of tumor necrosis factor (TNF) alpha and interleukin (IL) 6 from human PBMC stimulated by LPS were observed. Results Different concentrations of PGG and lipid A were combined and the reaction rate increased with the increase of concentration. The dissociation constant (Kd) value of the two was 5.2×10-7 mol/L. PGG has a strong neutralizing effect on LPS. PGG can significantly inhibit the release of TNF-α and IL-6 under the stimulation of LPS when the concentration is more than 20mg/L. At this time, the levels of TNF-α and IL-6 are from (1788±171) and (1966±232)ng, respectively. /L decreased to (958±234), (1351±99) ng/L (P<0.05), and this effect had a certain dose-effect relationship. Conclusion PGG can play a role in antagonizing LPS in vitro through the combination and neutralization of LPS.