骨髓间充质干细胞对大鼠急性肺损伤的影响  

Effect of bone marrow mesenchymal stem cells on acute lung injury in rats

作　　者：石智勇[1,2] 侯念果 李会[2] 帅训军[2] 杨洪光[2] 孙常荣[1] 路文卿 艾登斌 SHI Zhi-yong;HOU Nian-guo;LI Hui;SHUAI Xun-jun;YANG Hong-guang;SUN Chang-rong;LU Wen-qing;AI Deng-bin(Department of Anesthesiology,Weifang Medical College,Shandong Province,Weifang 261053,China;Department of Anesthesiology,Qingdao Municipal Hospital,Shandong Province,Qingdao 266011,China)

机构地区：[1]潍坊医学院麻醉学系,山东潍坊261053 [2]青岛市市立医院麻醉科,山东青岛266011

出　　处：《中国当代医药》2019年第29期11-15,共5页China Modern Medicine

基　　金：山东省医药卫生科技发展计划项目（2017WS209

摘　　要：目的探讨骨髓间充质干细胞(BMSCs)减缓大鼠急性肺损伤(ALI)的机制。方法36只6~7周龄雄性SD大鼠适应性饲养1周后,随机取12只作为对照组(Con组),其余大鼠均经尾静脉注射脂多糖(LPS)制作大鼠急性肺损伤模型。若大鼠模型的氧合指数(PaO2/FiO2)<300 mmHg,肺组织病理学评分升高,湿重(W)/干重(D)值升高,提示造模成功。将模型大鼠随机分为模型组(ALI组)和治疗组(BMSCs组),每组12只。BMSCs组造模1 h后气管内注射100μl BMSCs悬浮液(细胞总数为5×10^6),Con组和ALI组气管内注射100μl 0.9%生理盐水。24 h后腹主动脉置入导管抽取动脉血测定血氧分压(PaO2)并计算PaO2/FiO2;取肺组织行病理学评分、测量W/D;酶联免疫吸附试验(ELISA)检测支气管肺泡灌洗液(BALF)中肿瘤坏死因子-α(TNF-α)、白细胞介素-1α(IL-1α)、白细胞介素-10(IL-10)水平;采用血细胞计数仪测定中性粒细胞数(PMN);蛋白免疫印迹法(Western-Blot)测定肺组织中Bcl-2、Bax、Caspase-3蛋白的表达,并计算Bcl-2/Bax。比较三组大鼠的PaO2/FiO2、W/D、肺组织病理学评分、TNF-α、IL-1α、IL-10、PMN、Caspase-3、Bcl-2、Bax及Bcl-2/Bax水平。结果ALI组和BMSCs组的W/D、肺组织病理学评分、TNF-α、IL-1α、IL-10水平、PMN数目、Caspase-3蛋白、Bax蛋白的表达高于Con组,PaO2/FiO2、Bcl-2蛋白表达、Bcl-2/Bax低于Con组,差异均有统计学意义(P<0.05);BMSCs组的W/D、肺组织病理学评分、TNF-α、IL-1α、PMN数目、Caspase-3蛋白及Bax蛋白表达低于ALI组,IL-10水平、PaO2/FiO2、Bcl-2蛋白及Bcl-2/Bax高于ALI组,差异均有统计学意义(P<0.05)。结论BMSCs可抑制大鼠ALI模型的肺组织细胞凋亡,其机制可能与上调了肺组织中的Bcl-2蛋白和下调了Bax蛋白和Caspase-3蛋白的表达有关。Objective To explore the mechanism of bone marrow mesenchymal stem cells(BMSCs)in alleviating acute lung injury(ALI)in rats.Methods Thirty-six male SD rats aged 6-7 weeks were fed for one week.Twelve rats were randomly selected as control group(Con group),the other rats were injected with lipopolysaccharide(LPS)via caudal vein to establish acute lung injury model.If the oxygenation index(PaO2/FiO2)<300 mmHg,the pathological score of lung tissue increased,and the wet weight(W)/dry weight(D)value increased,suggesting that the model was successful.The model rats were randomly divided into model group(ALI group)and treatment group(BMSCs group),with 12 rats in each group.In BMSCs group,100 ml BMSCs suspension was injected into the trachea after 1 hour of modeling(the total number of cells was 5×10^6),100 ml 0.9%saline was injected into the trachea of Con group and ALI group.After 24 hours,the abdominal aorta was inserted with a catheter to extract arterial blood and the partial pressure of oxygen(PaO2)was measured and PaO2/FiO2 was calculated.The lung tissues were taken for pathological scoring and W/D was measured.The levels of tumor necrosis factor-α(TNF-α),interleukin-1α(IL-1α)and interleukin-10(IL-10)in bronchoalveolar lavage fluid(BALF)were detected by enzyme-linked immunosorbent assay(ELISA).The number of neutrophils(PMN)was measured by blood cell counting instrument.The expression of Bcl-2 protein,Bax protein and Caspase-3 protein in lung tissue was determined by Western-Blot,and the expression of Bcl-2/Bax was calculated.The levels of PaO2/FiO2,W/D,lung histopatho logical score, TNF-α, IL-1α, IL-10, PMN, Caspase-3, Bcl-2, Bax and Bcl-2/Bax were compared among the three groups. Results The W/D, lung histopathological score, TNF-α, IL-1α, IL-10, PMN number, Caspase-3 protein and Bax protein expression in ALI group and BMSCs group were higher than those in Con group, while PaO2/FiO2, Bcl-2 protein expression and Bcl-2/Bax protein expression were lower than those in Con group, the differences were statist

关 键 词：骨髓间充质干细胞 急性肺损伤 脂多糖 凋亡 

分 类 号：R563[医药卫生—呼吸系统]