目的探讨银杏叶提取物EGb761对神经调节素B(Neuromedin B,NMB)诱导的分娩期小鼠子宫平滑肌细胞中NF-κB活性和IL-6表达的影响。方法应用原代培养的、NMB受体(Neuromedin B receptor,NMBR)表达阳性的分娩期小鼠子宫平滑肌细胞,联合NMBR和核因子κB(Nuclearfactor kappa B,NF-κB)p65的RNA干扰技术、real-time PCR和磷酸化ELISA等方法,确定EGb761对NMB诱导的小鼠子宫平滑肌细胞中NF-κB和IL-6表达的影响。结果高浓度的EGb761(100 mg.L-1)组,NF-κB p65DNA结合活性明显低于对照组及低浓度组(P〈0.01),中、低浓度EGb组与对照组比较差异均无统计学意义。高浓度的EGb761预处理,可明显下调NMB诱导的子宫平滑肌细胞中NF-κB活性和IL-6的表达(P〈0.01),且二者的变化呈正相关(r=0.892,P〈0.01)。EGb761对NMB诱导的NF-κB活性和IL-6表达的调节作用可被NMBR及NF-κB基因沉默所阻断,且沉默两基因的阻断效率无差异。结论 EGb761可借助NMBR通路,抑制NF-κB活性和IL-6表达的上调,影响平滑肌细胞的活动。
Aim To investigate the effects of Ginkgo biloba extract 761(EGb761) on the activity of nuclear factor κB p65(NF-κBp65) and expression of interleukin-6(IL-6) induced by Neuromedin B(NMB) in primary cultured smooth muscle cells from the term.Methods The primary cultured smooth muscle cells with positive expression of NMB receptor(NMBR) from the term were prepared.The combination of NMBR and NF-κBp65 RNA interference(RNAi),real time PCR and Phosphorylation ELISA methods were used to study the effects of EGb761 on the activity of NF-κBp65 and expression of IL-6 induced by NMB in smooth muscle cells.Results The NF-κBp65 DNA binding activity was significantly lower in EGb761 group with high concentration(100 mg.L-1) than that in control group and low dose concentration group(P 0.01).However,there were not differences compared low or middle concentration groups with control group.The pretreated with high concentration of EGb761 could significantly reduce NF-κBp65 activity and IL-6 expression induced by NMB in pregnant smooth muscle cells(P 0.01),and these changes were significantly correlated(r = 0.892,P 0.01).Besides,the inhibiting role of EGb761 could be blocked by NMBR and NF-κB RNAi,respectively.Moreover,the blocking efficiency of two genes knockdown showed no significant difference.Conclusion EGb761 can inhibit NF-κBp65 activity and expression of IL-6 via NMBR pathway in primary cultured smooth muscle cells from the term.