中国组织工程研究 ›› 2025, Vol. 29 ›› Issue (31): 6642-6648.doi: 10.12307/2025.547

• 脐带脐血干细胞 umbilical cord blood stem cells • 上一篇    下一篇

人脐带间充质干细胞抑制脓毒症小鼠肺组织细胞的焦亡

何长良1,王  炎2,罗  玲2,刘  坚2   

  1. 1中山市古镇人民医院呼吸与危重症医学科,广东省中山市   528400;2湘潭市中心医院/湖南大学附属医院综合ICU,湖南省湘潭市   411100
  • 收稿日期:2024-06-18 接受日期:2024-08-31 出版日期:2025-11-08 发布日期:2025-02-19
  • 通讯作者: 刘坚,博士研究生,主治医师,湘潭市中心医院/湖南大学附属医院综合ICU,湖南省湘潭市 411100
  • 作者简介:何长良,男,1989年生,2013年长沙医学院毕业,主治医师,主要从事急性呼吸窘迫综合征、慢性阻塞性肺疾病等肺部急慢性疾病的诊疗与研究。
  • 基金资助:
    湖南省自然科学基金科卫联合项目(2023JJ60375),项目负责人:王炎;湖南省卫健委科研项目(D202317018303),项目负责人:罗玲

Human umbilical cord-derived mesenchymal stem cells thwart pyroptosis of lung tissue cells in septic mice

He Changliang1, Wang Yan2, Luo Ling2, Liu Jian2   

  1. 1Department of Respiratory and Critical Care Medicine, Guzhen People’s Hospital, Zhongshan 528400, Guangdong Province, China; 2Department of General Intensive Care Unit, Xiangtan Central Hospital/Affiliated Hospital of Hunan University, Xiangtan 411100, Hunan Province, China
  • Received:2024-06-18 Accepted:2024-08-31 Online:2025-11-08 Published:2025-02-19
  • Contact: Liu Jian, Doctoral candidate, Attending physician, Department of General Intensive Care Unit, Xiangtan Central Hospital/Affiliated Hospital of Hunan University, Xiangtan 411100, Hunan Province, China
  • About author:He Changliang, Attending physician, Department of Respiratory and Critical Care Medicine, Guzhen People’s Hospital, Zhongshan 528400, Guangdong Province, China
  • Supported by:
    Hunan Natural Science Foundation Science and Health Joint Project, No. 2023JJ60375 (to WY); Hunan Provincial Health Commission Research Project, No. D202317018303 (to LL)

摘要:

文题释义:

脐带间充质干细胞:是从胎儿分娩后脐带组织中分离培养获得的一种间充质干细胞,由于细胞来源为废弃的脐带组织,所以具有组织来源丰富、无创伤、伦理限制少的优点。此外,脐带间充质干细胞还具有良好的增殖、分化与免疫调节能力。 
脓毒症:是可导致机体严重器官功能障碍、危及生命的炎性综合反应,一般由病原微生物侵入机体引发。脓毒症的发病机制尚未完全阐明,近年研究认为免疫炎症因子风暴、免疫抑制、免疫麻痹和免疫耗竭带来的免疫功能紊乱是其重要病理机制,这些机制最终导致器官损伤和功能障碍。

摘要
背景:急性肺损伤是脓毒症患者的常见严重并发症,其发病机制复杂,目前尚无有效的药物治疗方案。人脐带间充质干细胞通过抑制细胞过度焦亡从而改善脓毒症急性肺损伤的作用机制愈发受到重视。
目的:探索人脐带间充质干细胞对脓毒症所致急性肺损伤小鼠肺组织细胞焦亡的影响及机制。
方法:48只Balb/c雄性小鼠按随机数字法分为假手术组、模型组、治疗组(n=16)。模型组、治疗组行盲肠结扎穿孔建立脓毒症所致急性肺损伤模型,假手术组仅开腹后缝合。治疗组术后6 h尾静脉注射200 μL人脐带间充质干细胞悬液(5×105个细胞),模型组及假手术组术后6 h尾静脉注射200 μL生理盐水。术后28 h,各组随机取5只小鼠予以尾静脉注射伊文思蓝检测肺血管通透性;收集各组剩余小鼠肺泡灌洗液和肺组织,ELISA检测肺泡灌洗液中白细胞介素1β、白细胞介素18水平,计数肺泡灌洗液中细胞总数及巨噬细胞数;苏木精-伊红染色观察肺组织病理学形态,TUNEL染色观察肺组织细胞焦亡情况,RT-PCR、Western blot检测肺组织中Toll样受体4、GSDMD及Caspase-11的mRNA和蛋白表达。
结果与结论:与模型组相比,治疗组术后28 h肺血管通透性显著下降(P < 0.05),肺泡灌洗液中白细胞介素1β、白细胞介素18水平降低(P < 0.05),细胞总数及巨噬细胞数降低(P < 0.05),肺损伤程度减轻,肺组织细胞焦亡指数降低(P < 0.05),肺组织Toll样受体4、GSDMD、GSDMD-N及Caspase-11 mRNA及蛋白表达水平均显著下降(P < 0.05)。结果表明,人脐带间充质干细胞可能通过抑制Toll样受体4/Caspase-11/GSDMD信号通路活化,改善肺组织细胞焦亡,从而在一定程度上改善脓毒症小鼠肺损伤的严重程度。

关键词: 人脐带间充质干细胞, Toll样受体4, Caspase-11, GSDMD, 急性肺损伤, 急性呼吸窘迫综合征, 细胞焦亡, 脓毒症, 工程化干细胞

Abstract: BACKGROUND: Acute lung injury is a common severe complication in sepsis patients, with a complex pathogenesis, for which there is currently no effective pharmacological treatment. The therapeutic mechanism by which human umbilical cord-derived mesenchymal stem cells improve sepsis-induced acute lung injury through the inhibition of excessive cellular pyroptosis is increasingly receiving attention.
OBJECTIVE: To investigate the effects and mechanisms of human umbilical cord-derived mesenchymal stem cells on cellular pyroptosis in lung tissues of mice with sepsis-induced acute lung injury.
METHODS: Totally 48 male Balb/c mice were randomly divided into sham, model, and treatment groups (n=16 per group). The model and treatment groups underwent cecum ligation and puncture to establish a sepsis-induced acute lung injury model, while the sham group underwent laparotomy without further procedures. The treatment group received a tail vein injection of 200 μL of human umbilical cord-derived mesenchymal stem cell suspension (5×105 cells) 6 hours after the procedure, while the model and sham groups received 200 μL of saline 6 hours after the surgery. Twenty-eight hours post-procedure, five mice from each group were randomly selected for tail vein injection of Evans blue dye to assess pulmonary vascular permeability. The remaining mice from each group had their bronchoalveolar lavage fluid and lung tissues collected for ELISA to measure levels of interleukin-1β and interleukin-18, cell counts, and macrophage numbers in the bronchoalveolar lavage fluid. Hematoxylin and eosin staining was used to observe the pathological morphology of lung tissue. TUNEL staining was used to assess cellular pyroptosis. RT-PCR and western blot analyses were conducted to measure mRNA and protein expression levels of Toll-like receptor 4, GSDMD, and Caspase-11 in lung tissues.
RESULTS AND CONCLUSION: Compared to the model group, the treatment group showed a significant decrease in pulmonary vascular permeability at 28 hours post-procedure (P < 0.05), reduced levels of interleukin-1β and interleukin-18 in bronchoalveolar lavage fluid (P < 0.05), and lower cell and macrophage counts (P < 0.05). Lung injury severity was reduced, with a decreased pyroptosis index (P < 0.05) and significantly lower levels of Toll-like receptor 4, GSDMD, GSDMD-N, and Caspase-11 mRNA and protein expression in lung tissues (P < 0.05). These results suggest that human umbilical cord-derived mesenchymal stem cells may improve the severity of sepsis-induced lung injury to some extent by inhibiting the activation of the Toll-like receptor 4/Caspase-11/GSDMD signaling pathway and reducing cellular pyroptosis in lung tissues. 

Key words: human umbilical cord-derived mesenchymal stem cell, Toll-like receptor 4, Caspase-11, GSDMD, acute lung injury, acute respiratory distress syndrome, pyroptosis, sepsis, engineered stem cell

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