穿心莲内酯对细菌内毒素所致脓毒症的体内外拮抗研究

Antagonism of Andrographolide Against Sepsis Induced by Bacterial Endotoxin in Vitro and in Vivo

目的:观察穿心莲内酯的体内外抗内毒素作用。方法:首先配置80、40和20μg/mL的穿心莲内酯浓度,分别加入10 ng/mL的内毒素温孵进行中和试验,利用鲎试剂定量测定内毒素含量,计算不同浓度的穿心莲内酯对内毒素的清除率,比较其体外抗内毒素的作用。其次配置80、40和20μg/mL的穿心莲内酯浓度,分别对10 ng/mL的内毒素刺激RAW264.7细胞所表达的相关因子进行抑制实验。最后配置160、80和40μg/mL的穿心莲内酯浓度,对内毒素攻击小鼠进行保护实验。结果:体外80、40和20μg/mL的穿心莲内酯浓度分别对内毒素中和试验对应的清除率为78.3%、30.8%和20.6%,差异有统计学意义;80、40和20μg/mL的穿心莲内酯浓度对RAW264.7细胞所表达的肿瘤坏死因子-α(TNF-α)、血清炎症因子白介素-6(IL-6)水平均有不同程度的降低,高浓度的穿心莲内酯降低程度最高;不同浓度的穿心莲内酯组能显著降低内毒素血症小鼠TNF-α和IL-6水平。结论:不同浓度的穿心莲内酯在体内外均有不同程度的抗内毒素作用,相对地,浓度越高抗内毒素效果更好。

Objective:To observe the antiendotoxin effect of andrographolide in vitro and in vivo.Method:Firstly,the concentration of andrographolide(80,40 and 20μg/mL),10 ng/mL endotoxin was added to incubate for neutralization test.The endotoxin concentration was measured quantitatively with TAL reagent,and the clearance rate of andrographolide of different concentrations on endotoxin was calculated to compare its antiendotoxin effect in vitro.Secondly,the concentration of andrographolide(80,40,and 20μg/mL)inhibited the expression of related factors in RAW264.7 cells stimulated by endotoxin(10 ng/mL).Finally,configuration andrographolide concentration(160,80,and 40μg/mL)was used to protect mice from endotoxin attack.Results:In vitro the clearance rates of andrographolide(80,40 and 20μg/mL)to endotoxin neutralization test were 78.3%,30.8%and 20.6%,respectively,with statistical difference;the concentration of andrographolide(80,40 and 20μg/mL)decreased the expression of TNF-αand IL-6 in RAW264.7 cells in varying degrees.The highest concentration of andrographolide decreased the levels of TNF-αand IL-6;it can significantly reduce the levels of TNF-αand IL-6 in mice with endotoxemia.Conclusion:Andrographolide of different concentrations in has different degrees of anti endotoxin effect in vivo and in vitro,relatively the higher the concentration,the better the anti endotoxin effect.