吸入骨髓间充质干细胞来源外泌体减轻慢性阻塞性肺疾病的炎性损伤  

Inhalation of bone marrow mesenchymal stem cells-derived exosomes alleviates inflammatory injury in chronic obstructive pulmonary disease

作　　者：王敏[1] 尹秀山 王盈熹 张妍[1] 赵龙[1] 夏书月[1] Wang Min;Yin Xiushan;Wang Yingxi;Zhang Yan;Zhao Long;Xia Shuyue(Department of Respiratory Medicine,Center Hospital Affiliated to Shenyang Medical College,Shenyang 110024,Liaoning Province,China;Shenyang University of Chemical Technology,Shenyang 110142,Liaoning Province,China)

机构地区：[1]沈阳医学院附属中心医院呼吸内科,辽宁省沈阳市110024 [2]沈阳化工大学,辽宁省沈阳市110142

出　　处：《中国组织工程研究》2023年第6期827-834,共8页Chinese Journal of Tissue Engineering Research

基　　金：辽宁省科学技术计划项目(LNCCC-D14-2015),项目负责人:夏书月;沈阳市科学技术计划项目(17-230-9-05),项目负责人:夏书月;沈阳市科学技术计划项目(20-205-4-073),项目负责人:赵龙;沈阳医学院硕士研究生科技创新基金项目(Y20210516),项目负责人:王敏。

摘　　要：背景:研究表明骨髓间充质干细胞源外泌体在多种呼吸系统炎症及疾病损伤模型中表现出强大的修复和再生能力,但在慢性阻塞性肺疾病中的研究较少,且尚未有研究将外泌体雾化吸入应用于慢性阻塞性肺疾病的模型实验中。目的:探讨大鼠骨髓间充质干细胞源外泌体通过雾化吸入途径对慢性阻塞性肺疾病大鼠炎症和肺部损伤的治疗作用,并且明确最适治疗剂量。方法:体外分离培养大鼠骨髓间充质干细胞,并提取鉴定其外泌体。脂多糖联合烟熏28 d建立慢性阻塞性肺疾病大鼠模型,然后给予低剂量(0.5×10^(8)particles/kg)、中剂量(1.0×10^(8)particles/kg)、高剂量(1.5×10^(8)particles/kg)外泌体雾化吸入治疗以及外泌体(1.5×10^(8)particles/kg)尾静脉注射治疗,模型组雾化1 mL PBS,对照组不造模、雾化1 mL PBS。连续雾化或注射治疗5 d,最后一次雾化或注射治疗后的第2天开始检测,使用小动物肺功能仪测试各组肺功能指标,ELISA检测支气管肺泡灌洗液及血清中的白细胞介素1β和肿瘤坏死因子α水平,苏木精-伊红染色和Masson染色从组织学评估肺组织改变。结果与结论:①骨髓间充质干细胞来源外泌体在透射电镜下显示为椭圆形的双层膜囊泡结构,呈典型的杯口状,粒径分析提示外泌体的峰直径为91.7 nm,占比为97.3%,颗粒浓度为3.3×10^(9) L^(-1),并且外泌体表面蛋白CD9和CD63高表达;②与尾静脉注射外泌体相比,雾化吸入外泌体显著改善了慢性阻塞性肺疾病大鼠的肺功能、肺组织切片胶原沉积和肺组织病理变化,并且明显降低了支气管肺泡灌洗液和血清中白细胞介素1β和肿瘤坏死因子α水平,且低剂量外泌体治疗效果最为显著;③以上结果表明,雾化吸入骨髓间充质干细胞来源外泌体可以减轻慢性阻塞性肺疾病的炎性损伤,并且最适剂量可能为0.5×10^(8)particles/kg。BACKGROUND:Many studies have demonstrated that exosomes derived from bone marrow mesenchymal stem cells show strong repair and regeneration ability in various models of respiratory inflammation and disease injury,but there are few studies on chronic obstructive pulmonary disease,and no studies have applied aerosolized inhalation of exosomes in model experiments of chronic obstructive pulmonary disease.OBJECTIVE:To investigate the therapeutic effects of exosomes derived from rat bone marrow mesenchymal stem cells on inflammation and lung injury in rats with chronic obstructive pulmonary disease by aerosol inhalation,and to determine the optimal therapeutic dose.METHODS:Rat bone marrow mesenchymal stem cells were isolated and cultured in vitro,and exosomes were extracted and identified.The rat model of chronic obstructive pulmonary disease was established by lipopolysaccharide combined with smoking for 28 days.Then low dose(0.5×10^(8)particles/kg),medium dose(1.0×10^(8)particles/kg),and high dose(1.5×10^(8)particles/kg)exosome aerosol treatment and exosome(1.5×10^(8)particles/kg)were given by tail vein injection.The model group was atomized with 1 mL PBS,while the control group was not molded with 1 mL PBS.Continuous atomization or injection was conducted for 5 days,and the test was started on the second day after the last atomization or injection treatment.The lung function indexes were tested by small animal pulmonary function instrument.The levels of interleukin-1βand tumor necrosis factor-αin bronchoalveolar lavage fluid and serum were detected by ELISA.The changes of lung tissues were assessed histologically by hematoxylin-eosin staining and Masson staining.RESULTS AND CONCLUSION:(1)The exosomes of bone marrow mesenchymal stem cells showed elliptic double-membrane vesicles under transmission electron microscopy,which were typical cup-shaped.Particle size analysis indicated that the peak diameter of exosomes was 91.7 nm,accounting for 97.3%,and the particle concentration was 3.3×10^(9) L^(-1).In addition

关 键 词：骨髓间充质干细胞 外泌体 慢性阻塞性肺疾病 雾化吸入 炎症 肺部损伤 

分 类 号：R459.9[医药卫生—治疗学] R318[医药卫生—临床医学] R563.3