吴茱萸碱对酵母多糖诱导急性腹膜炎小鼠的保护作用

陈浩; 范霞; 孙宇; 余静; 罗莉; 唐婉琦; 马娓; 杨雪; 梁华平; 龚建平

doi:10.16016/j.1000-5404.201608031

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吴茱萸碱对酵母多糖诱导急性腹膜炎小鼠的保护作用

基础医学 | 更新时间：2020-08-05

- 吴茱萸碱对酵母多糖诱导急性腹膜炎小鼠的保护作用
- Protective effect of evodiamine on zymosan-induced acute peritonitis in mice
- 第三军医大学学报 2017年39卷第2期页码：163-168
- 作者机构：
  
  1.400010 重庆, 重庆医科大学附属第二医院肝胆外科
  2.400042 重庆, 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
- 作者简介：
  
  龚建平, E-mail: gongjianping11@126.com Gong Jianping, E-mail: gongjianping11@126.com
- 基金信息：
  
  国家自然科学基金面上项目(81372088)
- DOI：10.16016/j.1000-5404.201608031
  中图分类号： R284.1;R392.3;R572.2
- 纸质出版日期：2017-1-30，
  
  收稿日期：2016-8-5，
  
  录用日期：2016-10-6
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陈浩, 范霞, 孙宇, 等. 吴茱萸碱对酵母多糖诱导急性腹膜炎小鼠的保护作用[J]. 第三军医大学学报, 2017,39(2):163-168.

Hao Chen, Xia Fan, Yu Sun, et al. Protective effect of evodiamine on zymosan-induced acute peritonitis in mice[J]. J Third Mil Med Univ, 2017,39(2):163-168.
陈浩, 范霞, 孙宇, 等. 吴茱萸碱对酵母多糖诱导急性腹膜炎小鼠的保护作用[J]. 第三军医大学学报, 2017,39(2):163-168. DOI： 10.16016/j.1000-5404.201608031.

Hao Chen, Xia Fan, Yu Sun, et al. Protective effect of evodiamine on zymosan-induced acute peritonitis in mice[J]. J Third Mil Med Univ, 2017,39(2):163-168. DOI： 10.16016/j.1000-5404.201608031.

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/mL ZYM溶液(1 mL)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/mL 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-kB (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(

＜0.05)，EVO治疗组上述指标均较模型组明显降低(

＜0.05)，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(

＜0.05)，且PMN中ROS显著降低(

＜0.05)。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(

＜0.05)。

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-kB p65入核实现的。

Abstract

Objective

To determine the effect of evodiamine (EVO) on zymosan(ZYM)-induced acute peritonitis in mice and explore its mechanism.

Methods

Fifty-four adult male C57BL/6 mice were randomly divided into control group (phosphate buffer saline+10% dimethyl sulphoxide)

model group (ZYM 50 mg/kg +10% dimethyl sulphoxide)

and EVO treatment group (EVO 10 mg/kg+ZYM 50 mg/kg)

and the drugs were administered by intraperitoneal injection. Six mice in each group were sacrificed at 2

6 and 12 h respectively after the injection. Blood samples and peritoneal lavage fluid (PLF) were collected. The levels of interleukin-6 (IL-6)

tumor necrosis factor-α (TNF-α)

keratinocyte-derived chemokine (KC) and macrophage inflammatory protein 2 (MIP-2) in the serum and PLF were measured by enzyme linked immunosorbent assay (ELISA). The numbers of leukocytes and polymorphonuclear neutrophil (PMN) in PLF were determined by hemocytometry and flow cytometry

respectively. The level of reactive oxygen species (ROS) in PMN was also detected by flow cytometry. The protein expression of nuclear factor-κB p65 (NF-κB p65) in the leukocytes was analyzed by Western blotting.

Results

After ZYM injection

the levels of IL-6

TNF-α

KC and MIP-2 in the serum and PLF were significantly elevated at 2

6 and 12 h (

＜0.05)

while EVO treatment could dramatically reduce the levels of all the cytokines and chemokines (all

＜0.05). The serum levels of IL-6

KC and MIP-2 reached the peak in 2 h and then declined gradually. While that of TNF-α was elevated in 2 h and got the summit in 6 h

followed by slow reduction. The PLF levels of above cytokines and chemokines were in similiar trends as in the serum. Furthermore

EVO could notably reduce the numbers of ZYM-induced inflammatory cells and PMN in the PLF(

＜0.05)

and reduce the percentage of PMN and its ROS level at 6 h(

＜0.05). Western blot results showed that EVO treatment could down-regulate the NF-κB p65 expression in peritoneal inflammatory cells.

Conclusion

EVO can effectively alleviate ZYM-induced peritoneal inflammatory injury in the mice through reducing the secretion of cytokines and chemokines

inhibiting the infiltration of leukocytes and neutrophils

decreasing the percentage of neutrophils and ROS production

and down-regulating NF-κB p65 expression in the nuclei.

关键词

吴茱萸碱急性腹膜炎酵母多糖中性粒细胞浸润

Keywords

evodiamineacute peritonitiszymosanpolymorphonuclear neutrophil infiltration

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