中文摘要：

目的探讨粒细胞-巨噬细胞集落刺激因子（GM-CSF）联合骨髓间充质干细胞（MSCs）对新生大鼠高体积分数氧（高氧）肺损伤的影响。方法采用贴壁选择法分离、培养、扩增大鼠骨髓来源MSCs,并以BrdU进行标记,注射前以PBS按要求剂量稀释;3日龄清洁级SD新生大鼠32只,置于950mL/L氧环境下7d,随机分为4组,每组8只;高氧加GM-CSF加mMSCs（联合组）,高氧加GM-CSF（GM-CSF组）,高氧加mMSCs（MSCs组）,高氧对照组（对照组）。联合组和GM-CSF组大鼠于模型结束后皮下注射GM-CSF9μg/kg,联合组同时腹腔注射5×104MSCs,MSCs组腹腔注射5×10^4MSCs,对照组仅注射相同容积的PBS。空气环境下再饲养3d。肺组织病理学检测其辐射状肺泡计数（RAC）;支气管肺泡灌洗液（BALF）细胞计数检测其白细胞,并进行细胞分类计数;ELISA法检测其肺组织匀浆TNF-α、GM-CSF水平。免疫组织化学法观察BrdU阳性细胞分布情况。结果1.4组RAC,BALF中白细胞、中性粒细胞、巨噬细胞,肺组织匀浆TNF-α、GM-CSF水平比较均有统计学差异（F=14.59P=0.000,F=5.65P=0.004,F=3.11P=0.042,F=26.31P=0.000,F=73.65P=0.000,F=43.91P=0.000）。与对照组比较,其他3组RAC值均明显增加（Pa〈0.05）、TNF-α水平均下降（Pa〈0.05）;GM-CSF组和联合组白细胞、中性粒细胞、巨噬细胞计数减少,GM-CSF水平增加（Pa〈0.05）,而MSCs组和对照组比较均无统计学差异（Pa〈0.05）。2.免疫组织化学显示联合组及MSCs组可见BrdU染色阳性细胞,联合组Br-dU积分高于MSCs组[（1.85±0.33）vs（1.04±0.18）分,t=6.09P=0.000]。结论GM-CSF、MSCs对高氧暴露引起的新生大鼠肺损伤均具有保护作用,可能与抑制损伤部位的炎性反应有关,而联合治疗具有协同作用。

英文摘要：

Objective To investigate the role of granulocyte macrophage-colony stimulating factor（GM-CSF） combined with mesenchymal stem cells （MSCs） on lung injury in hyperoxia-exposed newborn rats.Methods Mouse MSCs were isolated,cultured,amplified and labeled with 5-Bromo-2＇-deoxyuridine （BrdU）;After being cultured in 950 mL/L oxygen environment for 7 days,thirty-two 3-day-old Sprague Dawley（SD） rats were randomly divided into 4 groups as combined group,MSCs group,GM-CSF group and control group,and the rats were treated with MSCs（5×104 intraperitoneal injection） and（or） GM-CSF（9 μg/kg subcutaneous）,respectively;All animals were killed by an injection of sodium pentobarbital （120 mg/kg）.The value of radical alveolar counts （RAC）,the levels of TNF-α and GM-CSF in tissue homogenates,the count of white blood cells and the distribution of BrdU-positive cells were measured accordingly.Results 1.There were a significant differences in the field of value of RAC（F=14.59 P=0.000）,the levels of TNF-α（F=73.65 P=0.000） and GM-CSF（F=43.91 P=0.000）and the number of white blood cells （F=5.65 P=0.004） and neutrophils（F=3.11 P=0.042） and macrophage cells（F=26.31 P=0.000）.Compared with control group,the value of RAC was increased,while the levels of TNF-α were decreased in the others.While there were the increased levels of GM-CSF and the decreased numbers of white blood cells and neutrophils and macrophage cells in both GM-CSF group and combined group,there was no difference between MSCs group and control group in those fields.2.BrdU-positive cells were only found in combined group and MSCs group,and there existed significantly differences in BrdU-positive cells between the two groups[（1.85±0.33） vs （1.04±0.18） score,t=6.09 P=0.000].Conclusion GM-CSF and MSCs have synergistic protective roles against hyperoxia-induced lung injury in newborn rats,which may be related with the depression of inflammatory reaction of damaged region.