• 抑制Wnt/β-catenin信号通路可减轻脓毒症急性肺损伤
  • 熊杰.抑制Wnt/β-catenin信号通路可减轻脓毒症急性肺损伤[J].内科急危重症杂志,2021,27(6):511-516
    DOI:10.11768/nkjwzzzz20210616
    中文关键词:  脓毒症  急性肺损伤  Wnt/β-catenin信号通路  XAV939  修复
    英文关键词:
    基金项目:
    作者单位E-mail
    熊杰 华中科技大学同济医学院附属同济医院 908075933@qq.com 
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    中文摘要:
          摘要 目的:探讨抑制Wnt/β-catenin信号通路对脓毒症急性肺损伤(ALI)的影响。方法:选取(250±10)g健康成年SD雄性大鼠50只,随机分为3组:对照组10只、脓毒症ALI组(CLP组)20只、脓毒症ALI治疗组(CLP+XAV939组)20只。统计各组大鼠体重下降率及死亡率,取肺组织观察肺组织病理学变化,通过蛋白印迹法(WB)、免疫组化(IHC)、免疫荧光(IF)、实时定量PCR等技术检测Wnt/β-catenin信号通路关键蛋白(TNF-α、β-catenin、Cyclin D 、LRP5/6)表达在3组之间的差异性。结果:CLP组48h内死亡率为40%,而对照组和CLP+XAV939组无大鼠死亡。对照组大鼠术后48h体重无变化,而CLP组大鼠体重下降20%,CLP+XAV939组下降10%。组织病理学表现,对照组可见完整的肺泡结构,未见炎症细胞浸润;CLP组见肺泡水肿和塌陷,肺泡间隔增厚增宽并有大量炎症细胞浸润,肺血管充血明显;CLP+XAV939组可见基本完整的肺组织结构,炎症细胞数量及分部范围较CLP组减小。WB显示,与对照组比较,CLP组、CLP+XAV939组Wnt/β-catenin信号通路关键蛋白表达量上升,且CLP组高于CLP+XAV939组(P<0.01或P<0.05)。 实时定量PCR示,CLP组β-cateninmRNA表达量较对照组增高(P<0.05),CLP+XAV939组与对照组无差异。 IHC示,与对照组比较,CLP组和CLP+XAV939组肺内多种细胞内β-catenin蛋白表达量明显增多,且CLP组多于CLP+XAV939组(P<0.05)。IF示,与对照组比较,CLP组和CLP+XAV939组肺内LRP5/6表达量明显升高,且CLP组高于CLP+XAV939组(P均<0.05)。结论:Wnt/β-catenin信号通路参与脓毒症ALI过程,腹腔注射XAV939抑制Wnt/β-catenin信号通路可减轻ALI。
    英文摘要:
          Abstract Objective: To explore the effect of inhibiting the Wnt/β-catenin signaling pathway on acute lung injury in sepsis. Methods: A total of 50 SD male rats \[(250±10)g\] were randomly divided into three groups: control group (n=10), sepsis acute lung injury group (CLP group, n=20), and medication group (CLP+XAV939 group, n=20). The rate of weight loss and the mortality were observed. The pathological changes oflung tissue were examined. Western blotting, immunohistochemistry, immunofluorescence,and real-time quantitative PCR (RT-PCR) were used to investigate the difference in expression levels of the key proteins (TNF-α, β-catenin,CyclinD,LRP5/6) in the Wnt signaling pathway among three groups. Results: Within 48h, the mortality in the CLP group was 40%, and there were no deaths in the control group and the CLP+XAV939 group. After 48h, weight in the control group had no significant change, and that in the CLP group and the CLP+XAV939 group dropped by 20% and 10% respectively. Pathologically, the control group had intact alveolar structure with no inflammatory cell infiltration, the CLP group showed alveolar edema and collapse, thickened and widened alveolar septa and substantial inflammatory cells infiltration, and obvious pulmonary vascular congestion, and the CLP+XAV939 group had basically intact alveolar structure, with less infiltration of inflammatory cells and smaller distribution than the CLP group. The Western blotting showed that the expression of key proteins in the Wnt/β-catenin signaling pathway was increased significantly in the CLP group and CLP+XAV939 group as compared with the control group (P<0.01 or P<0.05),and that in the CLP group was higher than that in the CLP+XAV939 group(P<0.05). RT-PCR showed that the expression level of β-catenin mRNA in the CLP group was significantly higher than that in the control group (P<0.05), but there was no significant difference between the CLP+XAV939 group and the control group (P<0.05). The immunohistochemistry showed that the expression level of β-catenin in the CLP+XAV939 group and the CLP group was higher than that in the control group,and that in the CLP group was higher than that in the CLP+XAV939 group. The immunofluorescence showed that the expression level of LRP5/6 in the CLP+XAV939 group and the CLP group was higher than that in the control group, and that in the CLP group was higher than that in the CLP+XAV939 group. Conclusion: The Wnt/β-catenin signaling pathway is involved in the process of acute lung injury in sepsis.The intraperitoneal injection of XAV939 to inhibit Wnt/β-catenin signaling pathway can alleviate acute lung injury in sepsis.