白头翁汤及其主要成分对LPS诱导内皮细胞分泌NO、E-selectin和IL-8的影响

Effects of Pulsatillae Decoction and Its Main Ingredients on LPS-induced NO,E-selectin and IL-8 Secretion of Endothelial Cells

拟探讨白头翁汤及其主要成分对细菌内毒素(LPS)导致的机体炎症等病理损伤的治疗作用。培养内皮细胞至单层融合状态,加入内毒素刺激,3h后加入高、中、低3种浓度的8种药物,继续培养21h后收集细胞上清液,用ELISA方法测定NO、E-selectin和IL-8的含量。结果显示,白头翁汤高剂量组、白头翁素高剂量组和小檗碱高剂量组的NO含量极显著低于LPS对照组(P<0.01),白头翁汤中剂量组、白头翁素中剂量组、小檗碱中剂量组和秦皮乙素高剂量组显著低于LPS对照组(P<0.05);白头翁汤高剂量组和药根碱高剂量组的E-selectin含量极显著低于LPS对照组(P<0.01),白头翁皂苷B4高剂量组、小檗碱高剂量组和药根碱中剂量组显著低于LPS对照组(P<0.05);秦皮乙素高剂量组的IL-8含量极显著低于LPS对照组(P<0.01),白头翁汤高剂量组、秦皮甲素高剂量组和中剂量组、秦皮乙素中剂量组显著低于LPS对照组(P<0.05)。结果提示白头翁汤及其主要成分具有明显的抗细菌内毒素作用,可在由NO、E-selectin和IL-8介导的机体炎症等病理损伤过程中发挥治疗作用。

The aim of the research was to investigate the antiendotoxin of Pulsatillae Decoction （PD）, the levels of NO,E-selectin and IL-8 secreted by cultured rat intestinal microvascular endothelial cells were determined after treatment with its active ingredients, namely anemoside B4, anemonin, berberine, jatrorrhizine, palmatine, aesculin, and esculetin. Endothelial cell was challenged with 1 μg · mL^-1 LPS for 3 h, then treated respectively with PD at three concentrations （1, 5 and 10 mg · mL^-1） and its seven ingredients at three concentrations （1, 5 and 10 μg · mL^-1 ）. The cells were incubated at 37 ℃ in a cell incubator for 21 hours. The supernatants were collected and the content of NO,E-selectin and IL-8 were analyzed by ELISA kits. The results showed that as compared with LPS control group, the NO contents were significantly decreased in the middle- and high-dose groups of PD, the middle and high-dose groups of Anemonin, the middle- and high-dose groups of Berberine and the high-dose group of Esculetin （P〈0.01 or P〈0.05） ; The E-selectin contents were significantly decreased in the high-dose group of PD, the high-dose group of Anemoside B4, the high-dose group of Berberine and the middle- and high-dose groups of Jateorhizine （P〈0.01 or P〈0.05） ; The IL-8 contents were significantly decreased in high-dose group of PD, the middle- and high-dose groups of Aesculin and the middle- and high-dose groups of Esculetin （P〈0.01 or P〈0. 05）. It suggested that PD may exert beneficial therapeutic actions on systemic inflammatory response syndrome, multiple organ dysfunction syndrome and immune diseases mediated by NO, E-selectin and IL-8.