骨髓间充质干细胞对COPD-OSA重叠综合征大鼠肺血管内皮损伤的修复作用观察  

Repair effect of bone marrow mesenchymal stem cells on pulmonary vascular endothelial injury in rats with COPD-OSA overlap syndrome

作　　者：陈敏[1] 黄照明 郭翔[1] 毕虹[1] 杜俊毅[1] 杨超 杨丽娟 王丽艳[1] 邱云花 金志贤[1] CHEN Min;HUANG Zhaoming;GUO Xiang;BI Hong;DU Junyi;YANG Chao;YANG Lijuan;WANG Liyan;QIU Yunhua;JIN Zhixian(Kunming Calmette Hospital of the First People′s Hospital of Kunming,Kunming 650224,China)

机构地区：[1]昆明市第一人民医院附属甘美医院,昆明650224

出　　处：《山东医药》2020年第35期12-16,共5页Shandong Medical Journal

基　　金：国家自然科学基金资助项目(81560010);云南省应用基础研究计划项目[2017FE467(-100)];昆明市西山区科技计划项目(西科字22号)。

摘　　要：目的观察大鼠骨髓间充质干细胞(BMSCs)对慢性阻塞性肺疾病(COPD)-阻塞性睡眠呼吸暂停(OSA)重叠综合征大鼠肺血管内皮损伤的修复作用,并探讨其可能的机制。方法取SPF级雌性SD大鼠2只,分离、培养其BMSCs并进行鉴定。取SPF级雌性SD大鼠16只,采用烟熏联合间歇低氧的方法制备COPD-OSA重叠综合征大鼠模型,随机分为观察组和对照组,每组8只。观察组经尾静脉注射BMSCs 2×106/次,1次/周,共4次;对照组注射等体积、等次数的生理盐水。两组治疗后1周,处死后取肺组织。HE染色及MASSON染色后观察肺组织、肺小动脉病理改变,采用DAPI-CD34-TUNEL三重定性法观察肺组织血管内皮细胞凋亡情况,采用ELISA法检测肺组织内皮素1(ET-1)、内皮型一氧化氮合酶(eNOS)、血管内皮生长因子(VEGF)、基质细胞衍生因子1α(SDF-1α)表达。结果与对照组比较,观察组肺组织病理呈现的肺气肿、肺间质淋巴细胞浸润、中性粒细胞浸润以及支气管壁淋巴细胞增生、细支气管管壁平滑肌增生、杯状细胞增生等病理改变更轻,肺小动脉肌化、外膜胶原纤维沉积、血管壁结构异常、血管周围炎症、肺间质增厚等病理改变较轻。观察组与对照组肺组织血管内皮细胞凋亡率分别为2.3552%±0.3923%、1.3339%±0.1553%,两组比较P<0.01。观察组肺组织ET-1、VEGF表达均低于对照组,eNOS表达高于对照组(P均<0.05)。两组肺组织SDF-1α表达比较差异无统计学意义(P>0.05)。结论经尾静脉注射BMSCs可减轻COPD-OSA重叠综合征大鼠的肺组织血管内皮损伤、抑制血管内皮细胞凋亡,其机制可能与降低ET-1表达及升高eNOS表达有关。Objective To investigate the repair effect of bone marrow mesenchymal stem cells(BMSCs)on pulmonary vascular endothelial injury in rats with chronic obstructive pulmonary disease(COPD)-obstructive sleep apnea(OSA)overlap syndrome and to explore its possible mechanism.Methods BMSCs were isolated from femur and tibial bone marrow of SPF-grade female SD rats,and then were cultured and identified.Sixteen female SD rats with SPF grade were selected to prepare the RAT models of COPD-OSA overlap syndrome by smoking combined with intermittent hypoxia.The rats were randomly divided into the observation group and control group,with 8 rats in each group.The rats in the observation group were injected with 2×106 BMSCs per time,once a week,for 4 times in total,and the rats in the control group were injected with normal saline of the same volume.One week after treatment,the rats in the two groups were sacrificed,the lung tissues were taken for paraffin section for HE staining and MASSON staining to observe the pathological changes of the lung tissues and pulmonary arteriole,and the apoptosis of vascular endothelial cells in the lung tissues was observed by the triple qualitative method of DAPI-CD34-TUNEL,the expression levels of endothelin 1(ET-1),endothelial nitric oxide synthase(eNOS),vascular endothelial growth factor(VEGF)and stromal cell derived factor 1(SDF-1)were detected by ELISA.Results Compared with the control group,the pathological changes of the lung tissues in the observation group were milder,such as emphysema,interstitial lymphocyte infiltration,neutrophil infiltration,bronchial wall lymphocyte hyperplasia,bronchiolar wall smooth muscle hyperplasia and goblet cell hyperplasia;the changes of pulmonary arterioles myosization,outer membrane collagen fiber deposition,abnormal vascular wall structure,perivascular inflammation,and pulmonary interstitial thickening were relatively mild.The apoptosis rates of pulmonary vascular endothelial cells in the observation group and the control group were 2.3552%±0.3923%and 1

关 键 词：重叠综合征 慢性阻塞性肺疾病 阻塞型睡眠呼吸暂停 骨髓间充质干细胞 血管内皮损伤 损伤修复 大鼠 

分 类 号：R563.9[医药卫生—呼吸系统]