目的:银杏内酯B (GB)是已知的天然而强效的血小板激活因子受体拮抗剂.本实验通过研究GB对盲肠结扎穿孔(CLP)诱导的脓毒症小鼠的存活、外周血一氧化氮(NO)、活性氧簇(ROS)及细胞因子TNF-α、IL-1β、IL-6和IL-10的影响来评价其对脓毒症的免疫调节效果.方法:采用CLP的标准化程序建立小鼠脓毒症模型,术前30 min对药物组模型小鼠进行GB(10 μg/g)预处理;统计存活率;以Griess法检测小鼠外周血NO 水平;以H2DCFDA标记结合流式细胞术检测外周血ROS 水平;以流式细胞微球阵列法与ELISA法检测外周血细胞因子水平.结果:CLP手术后24 h,小鼠胸腺和脾脏萎缩明显,外周血炎症介质NO和ROS及细胞因子TNF-α、IL-1β、IL-6和IL-10的水平均显著上升,术后5 d内CLP模型小鼠全部死亡,而10 μg/g GB于术前30 min预处理,能缓解胸腺和脾脏萎缩,有效抑制炎症介质与炎性细胞因子风暴,显著提高脓毒症存活率.结论:GB预处理能显著降低CLP小鼠死亡率,下调外周血炎症介质NO和ROS及细胞因子TNF-α、IL-1β、IL-6和IL-10的水平,对脓毒症小鼠有明显的保护作用.
AIM: To explore the potential immunomodulatory effect and related mechanisms of ginkgolide B ( GB), a known potent antagonist of platelet - activating factor receptor, on the pathological process of sepsis. METHODS: The experimental sepsis model was established by a standardized procedure of cecal ligation and puncture (CLP). GB treatment (10 μg/g) was given to the CLP mice 30 min before the surgical operation. The survival rate was observed every day for 3 weeks. The NO content in the serum was measured by Griess assay. The ROS level in the blood was deter- mined by H2 DCFDA labeling and flow cytometry. The inflammatory cytokines in the serum were detected by the methods of cytometric bead array and ELISA. RESULTS: The thymus and spleen of the mice significantly atrophied, and the levels of NO, ROS, TNF -α, IL - 1β, IL - 6 and IL - 10 in the blood were dramatically elevated 24 h after CLP. All the CLP mice died in 5 days. However, treatment with 10 μg/g of GB 30 min before CLP remarkably enhanced the indexes of thymus and spleen, inhibited the storm of inflammatory mediators and cytokines, and improved the survival rate. CONCLUSION : The protective effect of GB on CLP - induced experimental sepsis indicates that GB is a candidate of natural immunomodulator for treating sepsis.