参附对脐静脉内皮细胞Toll样受体4/NF-κB途径的影响

目的观察参附注射液对内毒素介导的人脐静脉细胞Toll样受体4/NF-κB途径及细胞间黏附分子、E-选择素表达的影响。方法健康产妇分娩后的新鲜婴儿脐带,应用胶原酶消化,收集内皮细胞进行培养。实验分为对照组、内毒素组、内毒素+参附组。对照组:M199培养基中不加任何物质。内毒素组:M199培养基中加入脂多糖1 mg/L。内毒素+参附组:预先加入参附注射液10 mL/L,30 min后加入脂多糖1 mg/L。干预1h收集细胞,提取核蛋白测定NF-κB活性,干预12 h后收集细胞检测Toll样受体4、细胞间黏附分子1、E-选择素表达水平。结果内毒素组的Toll样受体4、细胞间黏附分子1、E-选择素表达水平、NF-κB活性均显著高于对照组,内毒素+参附组的Toll样受体4、细胞间黏附分子1、E-选择素表达水平、NF-κB活性均明显低于内毒素组。结论参附注射液可以通过抑制NF-κB活化,减弱Toll样受体4/NF-κB途径,抑制了脂多糖介导的黏附分子表达,从而减轻机体的炎症反应。

Effects of Shenfu injection on lipopolysaccharide-induce Toll-like receptor-(NF-κB) signal in human umbilical vein endothelial cells

Objective To investigate the effects of Shenfu injection on lipopolysaccharide(LPS)-induced Toll-like receptor-nuclear factor kappa B(TLR4/NF-κB) signal and intercellular adhesion molecule-1(ICAM-1),E-selection in human umbilical vein endothelial cells(HUVECs).Methods HUVECs obtained from collagenase-digested umbilical veins were cultured and divided into 3 group:①control group:cultured in M199 medium without any intervention.②LPS group:cultured in M199 medium with LPS(1 mg/L).③LPS+Shenfu group:pretreated with Shenfu injection for 30 min before LPS incubation.HUVECs were stimulated for 1 h,and NF-κB P65 activation were examined.HUVECs were stimulated for 12 h and then collected,TLR4,ICAM-1,and E-selection were examined.Results NF-κB P65 activity,TLR4,ICAM-1,E-selection mRNA and protein expression were higher in LPS group than in control group.NF-κB P65 activity,TLR4,ICAM-1 and E-selection mRNA and protein expression were lower in LPS+Shenfu group than in LPS group.Conclusion Shenfu injection play a role in inhibiting adhesion molecule up-regulation and inflammatory reaction by inhibiting NF-κB activation,and subsequently suppression TLR4/NF-κB signal.