骨髓间充质干细胞对小鼠肺脏缺血再灌注损伤的影响

　　[摘要]目的 探讨骨髓间充质干细胞对小鼠肺脏缺血再灌注损伤的影响。方法 采用全骨髓培养法分离纯化小鼠骨髓间充质干细胞（BMSCs），结扎左侧肺门60 min，再灌注240 min制备小鼠肺脏缺血再灌注损伤模型。70只小鼠随机分为7组（n=10），包括：假手术组（Sham组）、缺血再灌注组（I/R组）、BMSCs 1组（尾静脉注射1×106个BMSCs）、BMSCs 2组（尾静脉注射2×106个BMSCs）、BMSCs 5组（尾静脉注射5×106个BMSCs）、BMSCs 10组（尾静脉注射10×106个BMSCs）、BMSCs it组（气管内注射5×106个BMSCs）。再灌注结束时抽取小鼠股动脉血，行血气分析，并计算氧合指数（PaO2/FiO2）。随后处死小鼠，取肺组织，采用ELISA法检测定肺组织中超氧化物歧化酶（SOD）的活性、白细胞介素（interleukin，IL）-8、肿瘤坏死因子（TNF-α）含量及髓过氧化物酶（MPO）的活性，光镜下观察肺组织损伤程度并进行病理学评分。结果 与Sham组比较，I/R组、BMSCs 1组、BMSCs 2组、BMSCs 5组、BMSCs 10组、BMSCs it组的PaO2/FiO2降低，肺组织病理学评分、MPO活性、IL-8、TNF-α含量升高，SOD活性降低，差异有统计学意义（P<0.05）。与I/R组比较，BMSCs 1组、BMSCs 2组、BMSCs 5组、BMSCs 10组、BMSCs it组的PaO2/FiO2升高，肺组织病理学评分、MPO、IL-8、TNF-α水平降低，SOD活性升高，差异有统计学意义（P<0.05）；其中BMSCs 10组最明显，差异有统计学意义（P<0.05），气管内注射与血管内注射相同浓度BMSCs作用无明显区别，差异无统计學意义（P>0.05）。结论 BMSCs能够明显减轻肺组织缺血再灌注损伤，其作用机制与调节肺组织炎症反应有关，且具有明显量效关系，气管内注射与血管内注射相同浓度BMSCs作用无明显区别。
　　[关键词]骨髓间充质干细胞；肺；再灌注损伤；氧自由基
　　[中图分类号] R332 [文献标识码] A [文章编号] 1674-4721（2018）2（b）-0018-04
　　Influence of bone marrow mesenchymal stem cells on lung ischemia-reperfusion injury in mice
　　SUN Chang-rong1，2 HOU Nian-guo1 LI Hui1 SHUAI Xun-jun1 SUN Ming-jie1 TANG Yu-ru1 XU Tang-wen1，2 AI Deng-bin1▲
　　1.Department of Anesthesiology，Weifang Medical College，Shandong Province，Weifang 261053，China；2.Department of Anesthesiology，Research Center of Clinical Anesthesiology of Qingdao City，Qingdao Municipal Hospital，Shandong Province，Qingdao 266011，China
　　[Abstract]Objective To investigate the influence of bone marrow mesenchymal stem cells （BMSCs） on lung ischemia-reperfusion injury （LIRI） in mice.Methods BMSCs from rats were isolated，cultured and purified by the whole bone marrow acherence method.LIRI was produced by occlusion of the left hilum of lungs for 60 min followed by 240 min reperfusion.70 mice were randomly divided into 7 groups （n=10），included the sham operation group （the Sham group），the ischemia-reperfusion group （the I/R group），the BMSCs 1 group （tail vein injections of 1×106 BMSCs），the BMSCs 2 group （tail vein injections of 2×106 BMSCs），the BMSCs 5 group （tail vein injection of 5×106 BMSCs），the BMSCs 10 group （tail vein injection of 10×106 BMSCs），the BMSCs it group （endotracheal injection of 5×106 BMSCs）.At the end of reperfusion，blood samples were obtained for blood gas analysis and PaO2/FiO2 was calculated.Then，the animals were sacrificed and lung tissues were immediately removed for determination of superoxide dismutase （SOD），tumor necrosis factor-α （TNF-α），interleukin-8 （IL-8），and myeloperoxidase （MPO） and for microscopic examination of the pathological changes of lungs which were scored.Results Compared with Sham group，PaO2/FiO2 was significantly decreased，the pathological scores and the levels of TNF-α，IL-8，and MPO were increased，and SOD activity was decreased in I/R，BMSCs 1，BMSCs 2，BMSCs 5，BMSCs 10，BMSCs it group，the difference was statistically significant （P<0.05）.Compared with I/R group，PaO2/FiO2 was significantly increased，the pathological scores and the levels of TNF-α，IL-8，and MPO in lung tissues were decreased，and SOD activity was increased in BMSCs 1，BMSCs 2，BMSCs 5，BMSCs 10，BMSCs it group，the difference was statistically significant （P<0.05）.The most obvious group is BMSCs 10，the difference was statistically significant （P<0.05）.There is no obvious difference between endotracheal injection and intravascular injection with the same concentration of BMSCs，the difference was not statistically significant （P>0.05）.Conclusion BMCS can alleviate LIRI in mice，and the responsible mechanism is related to inhibition of inflammatory responses.The lung protective effect is dose dependent.There is no significant difference between endotracheal injection and intravascular injection with the same concentration of BMSCs.