SIRT1基因沉默在辐射诱导的NLRP3和IL-1β表达中的作用研究

Effects of NLRP3 and IL-1β on radiation-induced expression through SIRT1 gene silencing

    摘要
  • 摘要:
    目的 研究沉默信息调节因子1（SIRT1）基因沉默对间充质干细胞（MSCs）受照后Nod样受体蛋白3（NLRP3）和IL-1β表达的影响，探讨SIRT1激活剂——白藜芦醇的辐射防护作用及其机理。
    方法 将MSCs分为空白对照组、单纯照射组、RNA干扰组、白藜芦醇组和RNA干扰+白藜芦醇组。采用酶联免疫吸附测定、Western blot和RT-PCR等方法检测IL-1β、SIRT1和NLRP3在蛋白和mRNA水平的表达。
    结果 辐射可导致MSCs细胞外IL-1β分泌水平明显升高，给予白藜芦醇后，细胞IL-1β分泌水平较单纯照射组显著下降（t=21.68，P＜0.01），NLRP3和IL-1β mRNA水平较单纯照射组明显降低（t=14.44，P＜0.01；t=12.35，P＜0.01），SIRT1基因沉默后，NLRP3和IL-1β的mRNA水平回升至单纯照射组水平（t=14.86，P＜0.01；t=11.12，P＜0.01），即使再给予白藜芦醇，NLRP3和IL-1β的mRNA水平仍明显高于白藜芦醇组（t=11.31，P＜0.01；t=10.54，P＜0.01）。
    结论 SIRT1基因沉默减弱了白藜芦醇对辐射诱导的NLRP3和IL-1β的抑制作用，说明白藜芦醇可能通过激活SIRT1、抑制NLRP3、降低IL-1β的表达，从而减轻辐射引起的细胞损伤。

     

    Abstract:
    Objective To investigate the effect of silence information regulator-1(SIRT1) on NLRP3 and IL-1β production in mesenchymal stem cells(MSCs) exposed to radiation and to explore the SIRT1 role against radiation.
    Methods MSCs were divided into control, irradiation, RNAi, resveratrol, and RNAi+resveratrol group. IL-1β, SIRT1, and Nod-like receptor protein 3(NLRP3) expressions were detected using enzyme-linked immunosorbent assay(ELISA), Western blot analysis, and real-time PCR assay.
    Results Extracellular IL-1β secretion induced by radiation was increased significantly. After resveratrol treatment, the levels of IL-1β secretion decreased compared with those of the radiation group(t=21.68, P < 0.01). The mRNA level of NLRP3 and IL-1β also decreased compared with that of the radiation group(t=14.44, P < 0.01; t=12.35, P < 0.01). SIRT1 knockdown significantly suppressed resveratrol-mediated anti-inflammatory activity(t=14.86, P < 0.01; t=11.12, P < 0.01). The mRNA level of NLRP3 and IL-1β remained higher in the RNAi+resveratrol group than those in the resveratrol group (t=11.31, P < 0.01; t=10.54, P < 0.01).
    Conclusions SIRT1 gene silencing can suppress NLRP3 and IL-1β expression induced by radiation. Thus, resveratrol may alleviate cellular damage induced by radiation through activation of SIRT1, inhibition of NLRP3, and decrease in IL-1β expression.

     

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