薄荷酮对内毒素致炎症模型小鼠的保护作用研究

发布时间：2018-03-01 04:19

本文关键词： 薄荷酮内毒素炎症因子 NLRP炎症小体 cathepsin-B PXR　出处：《中国药理学通报》2017年02期 　论文类型：期刊论文

【摘要】：目的观察薄荷酮对内毒素(LPS)致炎症模型小鼠的保护作用,并初步探讨与NLRP3炎症小体激活相关的调控作用机制。方法♂C57BL/6J小鼠按体质量分层随机分为空白对照组、模型对照组、地塞米松组(5 mg·kg~(-1))、薄荷酮0.25 g·kg~(-1)及0.5 g·kg~(-1)剂量组。除地塞米松组实验当日腹腔注射给药1次外,其余各组小鼠连续灌胃给药5 d,每天1次。各组小鼠末次给药30 min后,除空白组小鼠外,其余各组小鼠腹腔注射LPS(15 mg·kg~(-1),10 m L·kg~(-1))制备炎症反应模型,造模12 h后,小鼠取血分离血清,采用ELISA及液相蛋白芯片技术测定炎症因子IL-18、IL-1β、IL-5、TNF-α、IFN-γ、粒细胞集落刺激因子(G-CSF)、粒细胞-巨噬细胞集落刺激因子(GM-CSF)及巨噬细胞炎性蛋白-1β(MIP-1β)的水平;取小鼠全血进行白细胞(WBC)计数;剖取肺组织计算肺脏指数,并测定肺组织中NO含量,进行肺组织病理组织学检查;RT-PCR法检测肺组织中NLRP3、caspase-1及IFN-α mRNA表达;免疫组化法观察肺组织中cathepsin B、P2X7R蛋白表达。结果 0.5 g·kg~(-1)薄荷酮明显降低内毒素致炎症模型小鼠血清IL-18、IL-1β、IL-5、TNF-α、IFN-γ、G-CSF、GM-CSF、MIP-1β的水平(P0.05或P0.01),下调模型小鼠肺组织NLRP3、IFN-α mRNA表达及cathepsin-B、P2X7R蛋白表达(P0.05或P0.01),明显降低模型小鼠肺组织中NO的含量(P0.05);0.25 g·kg~(-1)薄荷酮明显降低模型小鼠血清中IL-1β、IL-5、TNF-α、MIP-1β含量(P0.05),下调肺组织中P2X7R蛋白表达及NO水平(P0.05或P0.01);病理组织学检查显示0.5、0.25 g·kg~(-1)薄荷酮能明显减少肺组织切片中嗜中性粒细胞数量(P0.01),减轻肺泡膈的增厚;薄荷酮对模型小鼠全血白细胞计数与肺指数的升高表现出降低趋势。结论薄荷酮对内毒素致炎症模型小鼠有保护作用,能抑制血清多种炎性细胞因子的释放而减轻肺部炎性损伤,该作用可能与干扰NLRP3炎症小体的激活有关。
[Abstract]:Objective to observe the protective effect of menthone on inflammatory model mice induced by lipopolysaccharide (LPS), and to explore the regulatory mechanism related to the activation of NLRP3 inflammatory bodies. 鈾侰57BL / 6J mice were randomly divided into control group, model control group, dexamethasone group (5 mg 路kg ~ (-1)), menthol 0.25 g 路kg ~ (-1) and 0.5 g 路kg ~ (-1)). The other mice were given orally for 5 days, once a day. After the last administration for 30 min, the other groups were injected intraperitoneally with LPS(15 mg 路kg ~ (-1) and 10 mL 路kg ~ (-1) 路kg ~ (-1) LPS(15 for 30 min. After 12 hours, the serum samples were collected from the mice. ELISA and liquid-phase protein chip technique were used to detect the levels of IL-18, IL-1 尾, IL-5, TNF- 伪, G-CSF- 纬, G-CSF-, GM-CSFand macrophage inflammatory protein 1 尾 -MIP-1 尾, and WBCcount in whole blood of mice. Lung index was calculated and no content in lung tissue was measured. The expression of NLRP3caspase-1 and IFN- 伪 mRNA in lung tissue was detected by RT-PCR. Results 0. 5 g 路kg-1) menthone significantly decreased the level of IL-18, IL-1 尾, IL-5, TNF- 伪, IFN- 纬, G-CSFGM-CSFMIP-1 尾, P0.01, down-regulated the expression of NLRP3mIFN- 伪 IFN- 伪 and cathepsin-BP2X7R protein in lung tissue of model mice by immunohistochemical method, and down-regulated the expression of NLRP3- IFN- 伪 mRNA and cathepsin-BP2X7R protein by P05 or P0.01. P0.01A significantly decreased the content of no in the lung tissue of the model mice. (P0.05g 路kg-1) menthone significantly decreased the serum IL-1 尾 -IL-5TNF- 伪 -MIP-1 尾 content, down-regulated the expression of P2X7R protein and the level of no (P0.05 or P0.01A) in the lung tissue of the model mice, and the pathological examination showed that it was 0.5U 0.25 g 路kg-1) menthol could be used in the lung tissue of the model mice, and the expression of P2X7R protein and the level of no in the lung tissue were down-regulated (P0.05g 路kg-1, P0.05g 路kg-1). The number of neutrophilic granulocytes in lung tissue sections was significantly reduced (P 0.01), and the thickening of alveolar diaphragm was alleviated. Menthone showed a decreasing trend on the increase of whole blood leukocyte count and lung index in model mice. Conclusion menthone has protective effect on endotoxin-induced inflammatory model mice. It can inhibit the release of many inflammatory cytokines in serum and alleviate the pulmonary inflammatory injury, which may be related to interfering with the activation of NLRP3 inflammatory corpuscles.
【作者单位】：成都中医药大学药学院中药材标准化教育部重点实验室四川省中药资源系统研究与开发利用省部共建国家重点实验室培育基地;
【基金】：国家自然科学基金资助项目(No81473399) 国家基础科学人才培养基金项目(NoJ1310034-09)
【分类号】：R285.5;R-332

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