| 胆绿素对脂多糖诱导小鼠RAW264.7巨噬细胞NLRP3炎性体激活的作用 |
| 投稿时间：2017-01-03 修订日期：2017-02-01 点此下载全文 |
| 引用本文：龚睿,蒋磊,宋宁,费东生,杨松林,刘文,赵鸣雁.胆绿素对脂多糖诱导小鼠RAW264.7巨噬细胞NLRP3炎性体激活的作用[J].医学研究杂志,2017,46(10):35-39 |
| DOI：
10.11969/j.issn.1673-548X.2017.10.010 |
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| 基金项目:国家自然科学基金资助项目（81171785）；黑龙江省自然科学基金资助项目（ZJY0704-02）；哈尔滨医科大学研究生创新科研项目（YJSCX2015-21HYD） |
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| 中文摘要:目的 探讨胆绿素（biliverdin，BV）对脂多糖诱导RAW264.7巨噬细胞NLRP3炎性体激活的影响及其作用机制。方法 用不同浓度胆绿素（10、20、30μmol/L）处理小鼠RAW264.7巨噬细胞30min后再用LPS （1μg/ml）处理细胞6h，ELISA法检测培养上清液IL-β和IL-18中的分泌量。30μmol/L胆绿素处理细胞30min后再用LPS处理细胞6h，Western blot法检测胞内NLPR3、caspase-1、ASC和磷酸化IκB （p-IκB）的蛋白表达水平；NF-κB抑制剂BAY11-7082处理细胞1h后，然后给予细胞30μmol/L胆绿素孵育30min，经1μg/ml LPS孵育6h，ELISA法检测细胞培养上清液中IL-β、IL-18的表达水平。结果 （1）在受到LPS作用后，RAW264.7巨噬细胞分泌IL-β、IL-18水平显著升高；而在预先给予胆绿素后，抑制了LPS诱导的IL-β、IL-18表达，并且呈剂量依赖性（P < 0.05）。（2）与空白对照组相比，LPS处理巨噬细胞6h后，NLRP3炎性体各组分蛋白和p-IκB蛋白表达明显上调（P均<0.05），在同时间点，巨噬细胞在LPS处理前经过胆绿素预处理后，NLRP3炎性体各组分蛋白和p-IκB蛋白表达都显著降低（均P<0.05）。（3）与LPS组相比，BAY+胆绿素明显抑制了LPS诱导的IL-β、IL-18的表达（P<0.05），并且BAY和胆绿素的对于LPS诱导炎性反应对的联合抑制作用显著的高于BAY或者胆绿素的单独作用（P<0.05）。结论 在RAW264.7巨噬细胞中，胆绿素可以通过抑制NF-κB的活性进而抑制NLRP3炎性体的形成，从而发挥抗炎作用。 |
| 中文关键词:胆绿素 脂多糖 NLRP3炎性体 |
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| Effect of Biliverdin on NLRP3 Inflammasome Activation in Mouse Macrophage RAW264.7 Induced by Lipopolysaccharide and Its Mechanism |
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| Abstract:Objective To investigate the effects of biliverdin on NLRP3 inflammasome pathway in LPS-induced RAW264.7 macrophages. Methods RAW264.7 macrophage was pre-treated with different concentrations (10,20,30μmol/L) of biliverdin(BV) for 30 minutes, then administrated with LPS(1μg/ml) for 6 hours, and the expression of supernatant levels of proinflammatory cytokine IL-1β,IL-18 were measured respectively by ELISA. Cells were treated with LPS(1μg/ml) for 6h,and pretreated with BV(30μmol/L) for 30min before LPS stimulation, the protein expression of NLPR3,caspase-1,ASC and phosphorylated IκB were determined by western blot. Cells were preincubated with BAY11-7082 (an inhibitor of NF-κB) for 1 hour, followed by BV treatment and then LPS stimulation. Meanwhile, the levels of pro-inflammatory cytokine IL-1β, IL-18 in the supernatant were measured by ELISA. Results LPS significantly increased IL-1β, IL-18 scretion levels in the supernatant, however, pretreatment with BV suppressed the LPS-induced IL-1β, IL-18 in a concentration-dependent manner(P<0.05). The protein expressions of NLRP3 inflammasome and phosphorylated IκB were up-regulated significantly 6 hours after LPS stimulation compared with control group(P<0.05),while downregulated by BV on the same time point(P<0.05). Compared with LPS group, a large reduction in IL-1β, IL-18 expression were observed in BAY+BV+LPS group(P<0.05). Furthermore, these cytokines were much less in BAY+BV+LPS group than in BAY+LPS group or BV+LPS group(P<0.05). Conclusion Biliverdin exerted anti-inflammatory effects by regulating NLRP3 expression partially through the NF-κB pathway in RAW264.7 macrophages. |
| keywords:Biliverdin Lipopolysaccharide NLRP3 inflammasome |
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