骨髓来源间充质干细胞培养上清液减轻脂多糖诱导的小鼠急性肺损伤

发布时间：2018-11-05 16:59

【摘要】：目的:观察经尾静脉输注骨髓间充质干细胞培养上清液(MSCs CdM)对脂多糖(LPS)诱导的小鼠急性肺损伤的治疗作用及其机制。方法:采用全骨髓培养法分离纯化骨髓间充质干细胞,传至第3代时观察细胞形态,流式细胞术检测细胞表面标志,并且收集上清液用超滤离心管进行离心。30只BALB/c小鼠随机分为对照组、LPS模型组和MSCs CdM治疗组。对照组腹腔内注射生理盐水(0.01 mL/g),LPS组和MSCs CdM治疗组腹腔内注射LPS(5 mg/kg,0.01 mL/g)制备急性肺损伤模型。造模1 h后经尾静脉输注MSCs CdM(MSCs CdM治疗组)或生理盐水(LPS组或对照组)300μL。6 h后处死小鼠,留取标本检测肺组织病理形态学、肺组织湿干重比(W/D)、支气管肺泡灌洗液(BALF)中蛋白含量、血清及BALF中细胞因子水平和肺组织中髓过氧化物酶(MPO)的活性。结果:与对照组比较,LPS处理后肺组织病理损伤严重,BALF中蛋白、血清肿瘤坏死因子α(TNF-α)和白细胞介素6(IL-6)含量、肺组织中MPO活性及肺组织湿干重比均显著升高。与LPS组比较,MSCs CdM治疗组肺组织病理损伤程度减轻,BALF中蛋白、血清TNF-α和IL-6含量、肺组织中MPO活性及肺组织湿干重比均显著降低,而BALF中白细胞介素10(IL-10)和角质细胞生长因子(KGF)水平显著高于LPS组和对照组。结论:骨髓间充质干细胞培养上清液可有效减轻LPS诱导的急性肺损伤,其作用机制可能与其调节肺部TNF-α、IL-6、IL-10和KGF的水平有关。
[Abstract]:Aim: to observe the therapeutic effect of (MSCs CdM) on acute lung injury induced by lipopolysaccharide (LPS) in mice. Methods: bone marrow mesenchymal stem cells (BMSCs) were isolated and purified by whole bone marrow culture. Cell morphology was observed at the third passage, and cell surface markers were detected by flow cytometry. The supernatant was collected and centrifuged with ultrafiltration centrifuge tube. 30 BALB/c mice were randomly divided into control group, LPS model group and MSCs CdM treatment group. The model of acute lung injury was induced by intraperitoneal injection of normal saline (0. 01 mL/g), LPS) and intraperitoneal injection of LPS (5 mg/kg,0.01 mL/g) in the control group. The mice were sacrificed after 1 hour intravenous infusion of MSCs CdM (MSCs CdM or normal saline (LPS group or control group) for 1 hour. The lung tissue morphology and wet / dry weight ratio (W / D) of lung tissue were measured. Protein content in bronchoalveolar lavage fluid (BALF), cytokine level in serum and BALF and myeloperoxidase (MPO) activity in lung tissue. Results: compared with the control group, the pathological injury of lung tissue was serious after LPS treatment, and the contents of protein, serum tumor necrosis factor 伪 (TNF- 伪) and interleukin 6 (IL-6) in BALF were observed. The activity of MPO and the wet dry weight ratio of lung tissue increased significantly. Compared with LPS group, the pathological injury of lung tissue was alleviated in, MSCs CdM treatment group. The contents of BALF protein, serum TNF- 伪 and IL-6, the activity of MPO in lung tissue and the wet / dry weight ratio of lung tissue were significantly decreased in, MSCs CdM treatment group. The levels of interleukin 10 (IL-10) and keratinocyte growth factor (KGF) in BALF were significantly higher than those in LPS and control groups. Conclusion: the supernatant of bone marrow mesenchymal stem cell culture can effectively attenuate acute lung injury induced by LPS, and its mechanism may be related to the regulation of TNF- 伪, IL-6,IL-10 and KGF levels in lung.
【作者单位】：第四军医大学西京医院呼吸与危重症医学科;第四军医大学病理和病理生理教研室;
【基金】：国家自然科学基金资助项目(No.81072642;No.81372129)
【分类号】：R563.8

【参考文献】