吴茱萸碱对酵母多糖诱导急性腹膜炎小鼠的保护作用

发布时间：2018-03-30 02:18

本文选题：吴茱萸碱　切入点：急性腹膜炎　出处：《第三军医大学学报》2017年02期

【摘要】：目的探讨吴茱萸碱(evodiamine,EVO)对酵母多糖(zymosan,ZYM)诱导的小鼠腹膜炎中炎症反应及中性粒细胞(polymorphonuclear neutrophil,PMN)浸润的影响。方法将54只成年雄性C57BL/6小鼠按随机数字表法分为正常对照组、模型组和EVO治疗组。模型组腹腔注射1 mg/m L ZYM溶液(1 m L)建立急性腹膜炎小鼠模型,EVO治疗组则同时腹腔注射10 mg/kg EVO及1 mg/m L ZYM溶液,正常对照组腹腔注射等量磷酸盐缓冲液。各组处理后2、6、12 h处死6只小鼠,收集眼眶血及腹腔灌洗液。采用酶联免疫吸附法检测血清及腹腔灌洗液中白介素6(interleukin-6,IL-6)、肿瘤坏死因子α(tumor necrosis factor-α,TNF-α)、角化细胞来源趋化因子(keratinocyte-derived chemokine,KC)及巨噬细胞炎性蛋白2(macrophage inflammatory protein 2,MIP-2)的含量;通过血细胞计数板计数腹腔灌洗液中混合细胞总数;利用流式细胞仪检测腹腔灌洗液中PMN比例及PMN中活性氧(reactive oxygen species,ROS)水平;采用Western blot检测6 h时腹腔灌洗液混合细胞中核转录因子-k B(nuclear factor-κB,NF-κB)p65入核情况。结果建模后2、6、12 h,模型组血清及腹腔灌洗液中IL-6、TNF-α、KC及MIP-2含量均高于正常对照组(P0.05),EVO治疗组上述指标均较模型组明显降低(P0.05),其中,血清中IL-6、KC及MIP-2均于2 h达高峰,随后逐渐下降,而TNF-α在2 h逐渐上升,6 h达到高峰,随后逐渐下降;腹腔灌洗液中TNF-α、KC及MIP-2均于2 h达高峰,随后逐渐下降,而IL-6在2 h逐渐上升,6 h达到高峰,随后逐渐下降。制模后6 h,EVO治疗组可显著降低腹腔混合细胞数与PMN数及其百分比(P0.05),且PMN中ROS显著降低(P0.05)。另EVO治疗可显著降低腹腔混合细胞中细胞核的NF-κB p65表达(P0.05)。结论 EVO可抑制ZYM诱导急性小鼠腹膜炎模型中血清及腹腔灌洗液中炎症因子及趋化因子的分泌,减轻腹腔PMN的浸润,并抑制PMN中ROS的产生,该作用可能是通过抑制NF-k B p65入核实现的。
[Abstract]:Objective to investigate the effects of evodiamine EVOO on inflammatory response and polymorphonuclear neutrophil PMNs infiltration in mice peritonitis induced by yeast polysaccharide zymosanine. Methods 54 adult male C57BL/6 mice were randomly divided into normal control group. Model group and EVO group. The model group was treated by intraperitoneal injection of 1 mg/m L ZYM solution (1 mL) to establish acute peritonitis mouse model. The model group was treated with 10 mg/kg EVO and 1 mg/m L ZYM solution at the same time. In the normal control group, the same amount of phosphate buffer solution was injected intraperitoneally. Six mice were killed at 12 h after treatment in each group. The contents of interleukin-6 interleukin-6 (IL-6), tumor necrosis factor- 伪 (TNF- 伪), keratinocyte derived chemokine (KC) and macrophage inflammatory protein (2(macrophage inflammatory protein 2MIP-2) in serum and peritoneal lavage fluid were detected by Elisa. The total number of mixed cells in peritoneal lavage fluid was counted by blood cell counter, the ratio of PMN in peritoneal lavage fluid and reactive oxygen speciesrose in PMN were measured by flow cytometry. The nuclear transcription factor -k B(nuclear factor- 魏 B, NF- 魏 B)p65 in peritoneal lavage fluid mixed cells was detected by Western blot at 6 h. Results the levels of IL-6 TNF- 伪 KC and MIP-2 in serum and peritoneal lavage fluid in model group were higher than those in normal control group (P0.05EVO) at 12 h after modeling. Compared with the model group, the indexes were significantly lower than that of the model group. Serum IL-6 KC and MIP-2 reached their peak at 2 h, then decreased gradually, while TNF- 伪 gradually increased at 2 h and reached the peak at 6 h, then decreased gradually, and TNF- 伪 KC and MIP-2 in peritoneal lavage fluid reached the peak at 2 h and then decreased gradually. However, IL-6 gradually increased at 2 h and reached the peak at 6 h. After 6 h of model making, the number of mixed cells and the number and percentage of PMN in abdominal cavity were significantly decreased in EVO group, and the ROS in PMN decreased significantly (P 0.05). In addition, the expression of NF- 魏 B p65 in the nucleus of peritoneal mixed cells was significantly decreased by EVO treatment. Conclusion EVO can inhibit the secretion of inflammatory factors and chemokines in serum and peritoneal lavage fluid of acute peritonitis induced by ZYM in mice. It can reduce the infiltration of PMN in abdominal cavity and inhibit the production of ROS in PMN, which may be achieved by inhibiting the entry of NF-k B p65 into the nucleus.
【作者单位】：重庆医科大学附属第二医院肝胆外科;第三军医大学大坪医院野战外科研究所第一研究室创伤、烧伤与复合伤国家重点实验室;
【基金】：国家自然科学基金面上项目(81372088)~~
【分类号】：R259

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