银杏内酯B对脂多糖刺激的小鼠腹腔巨噬细胞TNFα生成及大鼠胸腔多形核白细胞NF-κB活化的影响

目的研究银杏内酯B对脂多糖刺激的小鼠腹腔巨噬细胞TNFα生成及大鼠胸腔多形核白细胞NF-κB活化的影响。方法用L929细胞结晶紫染色法检测TNFα的含量，用电泳迁移率改变检测法检测NF-κB的结合活性。结果1和10 μmol·L^-1银杏内酯B能够显著抑制LPS刺激的小鼠腹腔巨噬细胞TNFα的生成，其IC₅₀为0.26 μmol·L^-1；1 mg·L^-1 LPS和1 nmol·L^-1 PAF均可活化大鼠胸腔多形核白细胞NF-κB；银杏内酯B能够抑制LPS或 PAF刺激的NF-κB活化。结论银杏内酯B能够抑制LPS刺激的小鼠腹腔巨噬细胞TNFα生成及大鼠胸腔多形核白细胞NF-κB的活化。PAF参与LPS激活NF-κB的过程。

Effects of ginkgolide B on lipopolysaccharide-induced TNFalpha production in mouse peritoneal macrophages and NF-kappaB activation in rat pleural polymorphonuclear leukocytes]

AIM: To study the effects of ginkgolide B on lipopolysaccharide (LPS)--induced TNFalpha production in mouse peritoneal macrophages and NF-kappaB activation in rat pleural polymorphonuclear leukocytes. METHODS: L929 crystal violet staining assay was used to show the level of TNFalpha released from mouse peritoneal macrophages induced by LPS. Electrophoretic mobility shift assay (EMSA) was used to determine NF-kappaB binding activities. RESULTS: Ginkgolide B (1, 10 micromol x L(-1)) was shown to significantly inhibit LPS (10 mg x L(-1))-induced TNFalpha production in mouse peritoneal macrophages, the IC50 was 0.26 micromol x L(-1); LPS (1 mg x L(-1)) and PAF (1 nmol , L(-1)) were shown to increase the NF-kappaB binding activities in rat pleural polymorphonuclear leukocytes; ginkgolide B (10 micromol x L(-1)) was found to inhibit LPS (1 mg x L(-1))-induced NF-kappaB activation in rat pleural polymorphonuclear leukocytes; ginkgolide B (1, 10 micromol x L(-1)) was shown to inhibit PAF (1 nmol x L(-1))-induced NF-kappaB activation in rat pleural polymorphonuclear leukocytes. CONCLUSION: The inhibition of NF-kappaB activation and TNFalpha production might be considered to be part of the mechanisms underlying the antiinflammatory action of ginkgolide B; PAF is involved in activation of the NF-kappaB pathway stimulated with LPS.