| 宣伟,李帅,吴秀艳,耿艳,张卓.萝卜硫素对小鼠放射性肺损伤的防护作用机制[J].中华放射医学与防护杂志,2017,37(4):251-258 |
| 萝卜硫素对小鼠放射性肺损伤的防护作用机制 |
| Mechanism of protective effect of sulforaphane against radiation-induced lung injury in mice |
| 投稿时间：2016-11-30 |
| DOI：10.3760/cma.j.issn.0254-5098.2017.04.003 |
| 中文关键词: 萝卜硫素 放射性肺损伤 NLRP3 核转录因子-κB 白介素-1β |
| 英文关键词:Sulforaphane Radiation-induced lung injury NLR Family, Pyrin Domain-Containing 3 Protein Nuclear factor-κB Interleukin-1β |
| 基金项目:辽宁省科技厅肺癌转化医学研究中心建设项目（2014225003） |
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| 中文摘要: |
| 目的 探讨萝卜硫素（sulforaphane，SF）对小鼠急性放射性肺损伤的防护作用及其机制。方法 40只雌性C57BL/6J小鼠，按随机数字表法分为健康对照组、单纯照射组、照射+SF 3 mg/kg 组、照射+SF 5 mg/kg 组、照射+SF 10 mg/kg 组，每组8只。以6 MV X射线全肺单次12 Gy照射，各给药组自照射前7 d开始至照射后7 d，隔天腹腔注射不同浓度的SF，健康对照组和单纯照射组注射同等剂量的二甲基亚砜溶剂（DMSO+生理盐水）。照后14 d，留取小鼠肺组织标本，HE染色观察病理改变，免疫组织化学法观察NLRP3的定位与表达，ELISA法检测支气管肺泡灌洗液中IL-6、TNF-α、TGF-β1表达水平，实时荧光定量反转录聚合酶链反应（qRT-PCR）检测肺组织中NLRP3、IL-1β mRNA的表达，Western blot检测NF-κB p65、NLRP3、IL-1β蛋白表达，凝胶迁移实验（EMSA）检测NF-κB活性。结果 HE染色结果显示，各照射+SF组与单纯照射组比较，肺组织急性炎性反应减轻。肺组织中NLRP3表达下降，照射+SF 10 mg/kg组NLRP3降低显著（F=42.750，P<0.05）。支气管肺泡灌洗液中IL-6、TNF-α、TGF-β1水平下降（tIL-6=-62.65~-21.00，tTNF-α=-32.18~-16.57，tTGF-β1=-58.22~-46.11，P<0.05）。肺组织NLRP3和IL-1β mRNA表达显著下降（tNLRP3=-6.56~-5.68，tIL-1β=-29.75~-21.20，P<0.05）。NF-κB p65、NLRP3、IL-1β蛋白表达下降（tNF-κB p65=-34.00~-1.71，tNLRP3=-25.01~-16.91，tIL-1β=-73.70~-55.14，P<0.05），其中NF-κB p65、NLRP3相对表达量和SF呈剂量依赖性降低（r=0.945、0.926）。EMSA结果显示，NF-κB活性下降，且和SF呈剂量依赖性降低（tNF-κB=-38.68、-614.82、-2 831.40，P<0.05）。结论 萝卜硫素可通过降低小鼠肺组织NLRP3表达，有效地降低炎性因子的表达，减轻辐射诱导的炎症反应，对放射性肺损伤具有防护作用。 |
| 英文摘要: |
| Objective To investigate the radioprotective function and its mechanism of Sulforaphane (SF) in mice acute radiation-induced lung injury. Methods Totally 40 female C57BL/6J mice were equally divided into 5 groups randomly. Group A, treated by SF 3 mg/kg plus radiation; group B, treated by SF 5 mg/kg plus radiation; group C, treated by SF 10 mg/kg plus radiation; radiation group with a single dose of 12 Gy in 6 MV X-ray by a linear accelerator, and control group with sham radiation. The mice in drug group were administered intraperitoneally with different concentration of SF every other day from 7 d before irradiation to 7 d after irradiation, while the same volume of DMSO plus physiological saline solvent was given in the control and radiation groups. After being sacrificed at 14 d of SF administration, the pathomorphological changes of mice were observed in trauma lung tissue, the positioning and expression of NLRP3 was observed by immunohistochemical staining, the levels of IL-6, TNF-α and TGF-β1 in bronchoalveolar lavage fluid (BALF) were measured by ELISA, the expressions of NLRP3 and IL-1β mRNA in lung tissue were assayed by qRT-PCR, the expressions of NF-κB p65, NLRP3 and IL-1β proteins in lung tissue were assayed by Western blot, the activity of NF-κB was detected by EMSA. Results In comparison with radiation group, there was an obvious amelioration in pathological injury of lung tissue in the treatment groups: the expression of NLRP3 in lung tissue decreased; the concentration of NLRP3 in the drug intervention group (SF 10 mg/kg) markedly decreased (F=42.750, P<0.05). the IL-6, TNF-a and TGF-β1 levels in BALF decreased(tIL-6=-62.65-21.00; tTNF-α=-32.18-16.57; tTGF-β1=-58.22-46.11, P<0.05); the expressions of NLRP3 and IL-1β mRNA markedly decreased (tNLRP3=-6.56-5.68; tIL-1β=-29.75--21.20, P<0.05), and the expressions of NF-κB p65, NLRP3 and IL-1β proteins decreased (tNF-κB p65=-34.00--1.71, tNLRP3=-25.01--16.91, tIL-1β=-73.70--55.14, P<0.05); the relative expressions of NF-κB p65 and NLRP3 were reduced in a dose-dependent manner (r=0.945, 0.926); and the activity of NF-κB were obviously reduced (tNF-κB=-38.68, -614.82, -2 831.40, P<0.05). Conclusions Sulforaphane effectively alleviates the RILI in lung of mice by downregulating the expressions of inflammatory factor NLRP3. |
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