汪文婧,程储记,陈旭昕,韩志海.BMSC对内毒素诱导的肺泡上皮细胞氧化应激的影响[J].转化医学杂志,2021,10(4):206-211
BMSC对内毒素诱导的肺泡上皮细胞氧化应激的影响
Effects of Bone Marrow Mesenchymal Stem Cells on Lipopolysacharride Induced Alveolar Epithelial Cells in Oxidative Stress
  
DOI:
中文关键词:  肺泡上皮细胞  骨髓间充质干细胞  内毒素  急性呼吸窘迫综合症  氧化应激
英文关键词:Alveolar epithelial cell  Bone marrow mesenchymal stem cell  Lipopolysacharride  Acute respiratory distress syndrome  Oxidative stress
基金项目:国家自然科学基金资助项目 (81300050);中华国际医学交流基金会项目(Z-2017-24-2028-17)
作者单位
汪文婧 安庆市立医院心胸外科ICU 
程储记 安庆市立医院心胸外科ICU 
陈旭昕 中国人民解放军总医院第六医学中心呼吸与危重症医学科 
韩志海 中国人民解放军总医院第六医学中心呼吸与危重症医学科 
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中文摘要:
      目的 探讨骨髓间充质干细胞((bone marrow stromal cell,BMSCs)对内毒素(lipopolysaccharide, LPS)诱导的Ⅱ型肺泡上皮细胞(alveolar epithelial cellⅡ,AECⅡ)氧化应激的影响。方法 选用A549细胞代替AECⅡ,将实验分为A549组、A549+LPS组、A549-BMSC+LPS组及A549-BMSC+PBS组,即单独培养A549细胞或将BMSC与A549细胞共培养于Transwell体系,培养过夜后予LPS或等体积的PBS刺激。收集各组A549细胞样本,通过流式细胞仪检测活性氧(reactive oxygen species,ROS)的表达水平,并利用化学检测试剂盒,通过分光光度计检测脂过氧化物(lipid peroxides,LPO)、丙二醛(malondialdehyde,MDA)、超氧化物歧化酶(superoxide dismutase, SOD)及谷胱甘肽过氧化物酶(glutathione peroxidase,GSH)表达水平。结果 A549细胞在LPS的干预下,细胞内ROS表达水平(658.40±27.89)及其代谢产物LPO(1.46±0.15)、MDA(36.85±2.75)的水平明显升高(P<0.001),抗氧化因子SOD(13.44±3.32)、GSH(346.56±44.33)的活性明显降低(P<0.001);与A549+LPS组比较,A549-BMSC+LPS组ROS(544.61±41.30)、LPO(1.05±0.09)、MDA(10.63±0.95)的水平明显降低(P=0.001、P=0.006、P<0.001),SOD(33.41±3.59)、GSH(447.18±45.56)的活性明显升高(P=0.001、P=0.028);而与A549组比较,A549-BMSC+PBS组ROS(108.97±17.49)、LPO(0.12±0.03)、MDA(1.23±0.36)的表达水平呈下降趋势(P=0.033、P=0.004、P=0.012),SOD(68.33±8.01)、GSH(793.89±62.33)的活性进一步升高(P=0.001、P=0.002)。结论 BMSC能显著减轻LPS诱导的AECⅡ细胞的ROS生成,同时能正向调节其抗氧化的能力,可以一定程度上抑制LPS诱导的AECⅡ细胞的氧化应激反应。抑制肺泡上皮细胞氧化应激状态下ROS的产生、增强肺泡上皮细胞抗氧化的能力,或许可以成为治疗急性肺损伤/急性呼吸窘迫综合征的新策略之一。
英文摘要:
      Objective To investigate the effect of bone marrow mesenchymal stem cells (BMSC) on lipopolysaccharide (LPS) induced oxidative stress in alveolar epithelial cells (AECⅡ). Methods A549 cell was used as AECⅡin our study., and four groups including A549, A549+LPS, A549-BMSC+LPS and A549-BMSC+PBS were set. A549-BMSC means A549 was co-cultured with BMSC in the Transwell system. After overnight culture, they were stimulated with LPS or an equal volume of PBS. Then A549 was collected from each group, and the activity of reactive oxygen species (ROS) was measured with flow cytometry. Using the chemical detection kit, the levels of lipid peroxide (LPO), malondialdehyde (MDA), superoxide dismutase (SOD), and glutathione peroxidase (GSH) were measured with the spectrophotometer. Results Due to the intervention of LPS, compared to the A549 group, the levels of ROS (658.40±27.89), LPO (1.46±0.15), and MDA (36.85±2.75) in the A549+LPS group were significantly increased (P<0.001), while the levels of antioxidant factor SOD (13.44±3.32) and GSH (346.56±44.33) were significantly decreased (P<0.001). Compared to the A549+LPS group, the levels of ROS (544.61±41.30), LPO (1.05±0.09), and MDA (10.63±0.95) in the A549-BMSC+LPS group were significantly decreased (P=0.001, P=0.006, P<0.001), and the levels of SOD(33.41±3.59) and GSH(447.18±45.56) were significantly increased (P=0.001, P=0.028). Compared to the A549 group, the levels of ROS (108.97±17.49), LPO (0.12±0.03), and MDA (1.23±0.36) in the A549-BMSC+PBS group were also decreased (P=0.033, P=0.004, P=0.012), and levels of SOD (68.33±8.01), GSH (793.89±62.33) were increased (P=0.001, P=0.002). Conclusion BMSC could significantly reduce the ROS level in the AECⅡ induced by LPS, and improve its antioxidant capacity. BMSC could inhibit the oxidative stress response of AECⅡ induced by LPS. It may be a new treatment strategy for ALI/ARDS via inhibiting the production of ROS and enhancing the antioxidant capacity in the AECⅡ by BMSC.
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