实用医学杂志 ›› 2021, Vol. 37 ›› Issue (18): 2339-2343.doi: 10.3969/j.issn.1006⁃5725.2021.18.007

• 基础研究 • 上一篇    下一篇

血红素加氧酶⁃1修饰的骨髓间充质干细胞减轻脂多糖诱导的肺微血管内皮细胞氧化应激

陈旭昕1,2 李虎明1,2 唐俭1 孟激光1,2 韩志海1,2   

  1. 1 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(北京 100048);2 中国人民解放军总医院呼吸 与危重症医学部(北京 100091)

  • 出版日期:2021-09-25 发布日期:2021-09-25
  • 通讯作者: 韩志海 E⁃mail:zhihaihandoctor@163.com
  • 基金资助:
    国家自然科学基金资助项目(编号:81300050);北京市自然科学基金项目(编号:7182163);海军总医院创新培育基金项目(编号:CXPY201417);解放军总医院第六医学中心创新培育基金(编号:CXPY201902)

Bone marrow ⁃derived mesenchymal stem cells modified with heme oxygenase ⁃ 1 attenuate lipopolysaccha⁃ ride ⁃ induced oxidative stress in pulmonary microvascular endothelial cells

CHEN Xuxin*,LI Huming TANG Jian,MENG Jiguang,HAN Zhihai.   

  1. Department of Pulmonary and Critical Care Medicine,the Sixth Medical Center,PLA General Hospital,Beijing 100048,China;*College of Pulmonary and Critical Care Medicine,PLA General Hospital,Beijing 100091,China

  • Online:2021-09-25 Published:2021-09-25
  • Contact: HAN Zhihai E⁃mail:zhihaihandoctor@163.com

摘要:

目的 探讨血红素加氧酶⁃1(HO⁃1)修饰的骨髓间充质干细胞(MSC⁃HO⁃1)对脂多糖(LPS 刺激下肺微血管内皮细胞(PVEC)氧化应激的影响。方法 实验分组:A 组(PVEC +LPS),PVEC 仅予 LPS 刺激;B 组(PVEC/MSC+LPS),PVEC 细胞与 MSC 共培养于 Transwell 体系过夜后予 LPS 刺激;C 组(PVEC/ MSC⁃HO⁃1+LPS),同上 PVEC MSC⁃HO⁃1 共培养后予 LPS 刺激;D 组(正常 PVEC)。采用流式细胞仪及活 性氧检测试剂盒,测 PVEC 内活性氧(ROS)含量;采用比色法及相应商品化检测试剂盒,测 PVEC 内脂质过 氧化物(LPO)、丙二醛(MDA)水平及超氧化物歧化酶(SOD)、谷胱甘肽过氧化物酶(GSH⁃PX)活性;用 Western blot 检测各组 PVEC HO⁃1 表达变化。结果 D 组相比,A 组受 LPS 刺激后,出现明显氧化应激 P < 0.05);与 MSC MSC⁃HO⁃1 共培养均可明显减轻氧化应激(P < 0.05),但 MSC⁃HO⁃1 共培养减轻氧 化应激程度更为显著(P < 0.05)。Western blot 结果显示与 MSC⁃HO⁃1 共培养可明显上调 PVEC HO⁃1 白表达。结论 MSC⁃HO⁃1 显著改善 LPS 诱导的 PVEC 氧化应激失衡,可能机制与上调 PVEC HO⁃1 表达 有关。

关键词:

间充质干细胞, 脂多糖, 血红素加氧酶?1

Abstract:

Objective To explore the influence of bone marrow⁃derived mesenchymal stem cells modified with heme oxygenase⁃1(MSC⁃HO⁃1)on oxidative stress in pulmonary microvascular endothelial cells(PVECs following lipopolysaccharide(LPS)stimulation. Methods PVECs were divided into group A(PVEC + LPS), group B(PVEC/MSC+LPS),group C(PVEC/MSC⁃HO⁃1+LPS)and group D(PVEC). In groups B and C,PVECs were co⁃cultured with MSC or MSC⁃HO⁃1 overnight. LPS(0.1 mg/mL),as a stimulus,was thenadded into transwell co⁃culture system for 6 hours. Intracellular accumulation of ROS was measured using a commercial kit and flow cytometry. Activities of lipid peroxide(LPO),malondialdehyde(MDA),superoxide dismutase(SOD)and gluta⁃ thione peroxidase(GSH⁃PX)in PVECs were determined by using colorimetric assay. HO⁃1 protein expression of PVEC was analyzed by Western blot. Results As compared with group D,LPS⁃stimulation caused the imbalance of oxidative stress in PVECs. Co⁃culture with MSC or MSC⁃HO⁃1 significantly inhibited oxidative stress induced by LPS in PVECs,and the extent of inhibition in group C was more evident. Western blot analysis showed that co⁃ culture with MSC⁃HO⁃1 could significantly upregulate protein expression of HO⁃1 in PVECs. Conclusions MSC⁃ HO⁃1 attenuates lipopolysaccharide⁃induced oxidative stress in PVECs. The detailed mechanism might be associated with upregulation of HO⁃1 expression in PVECs.

Key words:

mesenchymal stem cell, lipopolysaccharide, heme oxygenase?1, oxidative stress, pul?monary microvascular endothelial cell