骨髓间充质干细胞不同时相干预对SD大鼠急性肺损伤的修复作用  

The effects of bone marrow mesenchymal stem cells intervention at different times on acute lung injury repairmen

作　　者：叶永顺 廖画 李文丰[1] 王依[1] 刘华[1] 

机构地区：[1]广东药科大学附属第一医院干保科,广东广州510080 [2]南方医科大学第五附属医院呼吸内科,广东广州510900

出　　处：《广东医学》2017年第5期657-661,共5页Guangdong Medical Journal

基　　金：广东省财政技术研究开发与推广应用专项基金项目(编号:粤财工[2013]401号)

摘　　要：目的探究骨髓间充质干细胞(BM-MSCs)不同时间干预对以静脉注射脂多糖(LPS)构建大鼠急性肺损伤(ALI)模型的修复作用。方法体外分离、培养并鉴定大鼠BM-MSCs。将120只雄性SD大鼠纳入研究,分成对照组、LPS组和LPS+BM-MSCs组,上述3组因BM-MSCs处理时间差异分成2、8、24、48、96 h共5个亚组(n=8)。LPS组以5 mg/kg尾静脉注射LPS,LPS+BM-MSCs组在注射LPS后分别在上述不同时间点尾静脉注射1 mL的BM-MSCs(1×10~6·mL^(-1));对照组则在上述时相点注射等剂量的生理盐水。检测各组肺泡灌洗液肿瘤坏死因子-α(TNF-α)、白细胞介素(IL)^(-1)α等炎症因子浓度、肺湿干比及肺损伤病理评分。结果成功分离、培养、纯化BM-MSCs,并诱导向成骨、成软骨、成脂细胞分化;流式细胞术鉴定所培养细胞系CD90、CD105、CD45、CD34阳性率分别为98.89%、97.37%、3.17%及1.41%。在LPS诱导的ALI中,TNF-α、IL^(-1)α、IL^(-1)0在8 h亚组中测得最高值[分别为(364.911±32.182)pg/mL、(286.316±18.567)pg/mL、(162.037±22.377)pg/mL],随着时间延长逐渐下降;肺湿干比在2 h亚组中测得最高值(6.361±0.265);肺水肿及组织损伤程度在2 h亚组中最严重,病理评分也测得最高值(0.462±0.048),随着时间延长逐渐下降。在LPS+BM-MSCs组中,2、8、24 h的炎症因子、湿干比及病理评分均有显著改变(P<0.05),随着时间延长,差异无统计学意义(P>0.05)。结论早期应用BM-MSCs可减少TNF-α、IL^(-1)α等炎症因子,从而减轻肺组织损伤及肺泡水肿。Objective To investigate the intervention effects of bone marrow - derived mesenchymal stem cells （ BM - MSCs） on the repairmen of acute lung injury （ALI） at different times. Methods The rat BM - MSCs were isola- ted, cuhured and identified in vitro. 120 male SD rats were divided into control group, LPS group and LPS ＋ MSCs group （ The above three groups were divided into 5 subgroups （2 h, 8 h, 24 h, 48 h, 96 h, n = 8 ） according to different time points of MSCs administration） ; in which placebo saline, lipopolysaccharide （LPS） , and LPS plus MSCs treatment were given, respectively. The inflammation factors, including TNF-α, IL -1α and et al. , in bronchoalveolar lavage fluid, the lung wet/dry ratio, and pathological score of lung injury were compared. Results With the cells isolated, cultured and further purified, the BM - MSCs can be induced to differentiate to adipocyte - oriented, chrondroeyte - oriented and osteocyte - oriented cells. The positive rates of CD90, CD105, CD45 and CD34 were respectively 98.89% , 97. 37% , 3. 17% and 1.41% by flow cytometry. The maximum concentration ofTNF -α, IL -1α, IL - 10 were observed in 8 h subgroup of LPS group [ （364. 911±32. 182） pg/mL, （286. 316±18. 567） pg/mL, （ 162. 037 ± 22. 377） pg/mL], and gradually decreased over time, while the maximum lung wet/dry ratio and pathological grading were observed in 2 h sub- group of LPS group（6. 361±0. 265, 0. 462 ± 0. 048 ）, then gradually decreased over time. As early intervention （2 h, 8 h, 24 h） of BM - MSCs, the inflammatory factor, dry wet ratio and pathological score were significantly different （P 〈 0. 05 ）, while there were no significant difference over time （ P 〉 0. 05 ）. Conclusion Early application of BM - MSCs can reduce TNF - α and IL - 1α, so as to reduce the lung injury and alveolar edema.

关 键 词：骨髓间充质干细胞 急性肺损伤 炎症因子 

分 类 号：R329.2[医药卫生—人体解剖和组织胚胎学]