LPS对小鼠枯否细胞核因子-κB活性的促进效应

STIMULATING EFFECT OF LPS ON THE ACTIVITY OF NF-κB IN MOUSE KUPFFER CELLS

为探讨细菌胞壁主要内毒素脂多糖 (Lipopolysaccharide ,LPS)对与炎症反应紧密相关的核因子 -κB(NF -κB)活性的影响 ,从正常对照和不同剂量的LPS刺激后不同时相的小鼠中分离出其肝脏枯否细胞 (Kupffercell,KC)。用凝胶迁移率改变分析法 (Electrophoreticmobilityshiftassay ,EMSA)检测KC核提取物中NF -κB的活化与LPS的量效、时效关系。结果发现 :刺激 3h后 ,低剂量的 (1mg/kg)LPS ,即可检测到NF -κB活性 ;中剂量组 (5mg/kg)的NF -κB活性达峰值 ,高剂量组 (10mg/kg)则仍可维持NF -κB的活化状态。中剂量 (5mg/kg)刺激 0 .5h即可检测到NF -κB活性 ,3h时NF -κB活性达峰值 ,并可持续到至少 8h。表明抑制NF -κB的过度活化可能有助于炎症的治疗。

To investigate the effects of lipopolysaccharides (LPS), i.e. the main endotoxic components of Gram-negative bacteria, on the inflammation-related nuclear factor-κB (NF-κB) activity in mouse kupffer cells, mice were divided into the control group and the LPS stimulated group, and electrophoretic mobility shift assay (EMSA) was performed. It was found that after 3 h of LPS stimulation the activity of NF-κB was detected in dosage 1 mg/kg LPS, reached the maximum in 5 mg/kg LPS and the latter level persisted in 10 mg/kg LPS. Stimulated with 5 mg/kg LPS, the DNA-binding activity of NF-κB was observed half an hour after LPS infusion, peaked at 3 h and persisted for at least 8 h. The activation of NF-κB by LPS observed in this experiment well explained the inflammatory effects of LPS and showed that inhibiting NF-κB activation might be useful in the therapy of inflammation.