骨髓间充质干细胞移植修复慢性阻塞性肺病气道损伤

Bone marrow mesenchymal stem cell transplantion for repair of airway injury in chronic obstructive pulmonary disease

背景:间充质干细胞能够分化为肺实质细胞并参与肺部损伤的修复,为间充质干细胞在慢性阻塞性肺病中的应用提供了新的方法。目的:观察骨髓间充质干细胞移植对大鼠慢性阻塞性肺病气道损伤的修复作用。方法:将24只雌性大鼠随机分为4组:1骨髓间充质干细胞移植组(12只):采用熏烟+脂多糖法建立慢性阻塞性肺病大鼠模型,于造模后第1天经尾静脉输注1 mL CM-Dil标记的骨髓间充质干细胞。2骨髓间充质干细胞对照组(4只):在第1天和第14天经气管注入生理盐水300μL,经尾静脉输注1 mL CM-Dil标记的骨髓间充质干细胞。3慢性阻塞性肺病模型组(4只):采用熏烟+脂多糖法建立慢性阻塞性肺病大鼠模型,于造模后第1天经尾静脉输注1 mL PBS。4健康对照组(4只):在第1天和第14天经气管注入生理盐水300μL,经尾静脉输注1 m L PBS。在各组大鼠注射骨髓间充质干细胞后的第1,7,15,30天,进行病理学和血清学检测。结果与结论:1苏木精-伊红染色结果显示:骨髓间充质干细胞移植组大鼠肺气肿和气道病变较慢性阻塞性肺病模型组轻,但较骨髓间充质干细胞对照组和健康对照组严重。2移植后第1天,骨髓间充质干细胞移植组外周血白细胞总数和中性粒细胞比例高于骨髓间充质干细胞对照组和健康对照组(P<0.05),随着时间延长,骨髓间充质干细胞移植组白细胞总数和中性粒细胞比例不断降低。3移植后第1天,骨髓间充质干细胞移植组外周血白细胞介素10水平低于骨髓间充质干细胞对照组和健康对照组(P<0.05),肿瘤坏死因子α和粒细胞集落刺激因子水平高于骨髓间充质干细胞对照组和健康对照组(P<0.05)。随着移植时间的延长,骨髓间充质干细胞移植组外周血肿瘤坏死因子α水平不断降低,白细胞介素10水平不断升高,粒细胞集落刺激因子水平先升高再降低,其中第7天水平最高。4CM-Dil染色联合免疫组化检测提示,部分CM-Dil阳性细胞同时CC16表达阳性。结果显示经尾静脉注射骨髓间充质干细胞能够改善慢性阻塞性肺病大鼠的肺部病理损伤,通过分化为气管黏膜上皮细胞以及参与免疫调节对气道进行修复。

BACKGROUND： Mesenchymal stem cells can differentiate into lung parenchymal cells involved in lung injury repair, providing a new approach for the application of mesenchymal stem cells in patients with chronic obstructive pulmonary disease.OBJECTIVE： To observe the effect of bone marrow mesenchymal stem cell transplantation on the repair of airway injury in rats with chronic obstructive pulmonary disease. METHODS： Twenty-four female rats were randomized into four groups： bone marrow mesenchymal stem cell transplantation group（cell transplantation group, n=12）; bone marrow mesenchymal stem cells group（cell controlgroup, n=4）; model group（n=4）; healthy control group（n=4）. Rat models of chronic obstructive pulmonary disease were established in the cell transplantation group and model group using fumigation＋lipopolysaccharide method; and at 1 day after modeling, model rats were given 1 mL CM-Dil-labeled bone marrow mesenchymal stem cells and 1 mL PBS via the tail vein in these two groups, respectively. In addition to tracheal injection of normal saline（300 μL） at 1 and 14 days, rats in the cell control and healthy control groups were given 1 mL CM-Dil-labeled bone marrow mesenchymal stem cells and 1 mL PBS via the tail vein, respectively. At 1, 7, 15 and 30 days after cell transplantation, lung tissue and serum markers of all rats were detected. RESULTS AND CONCLUSION：（1） Hematoxylin-eosin staining showed that emphysema and airway injury was milder in the cell transplantation group than the model group, but severer than the cell control and healthy control groups.（2） The total number of leukocytes and neutrophils in the peripheral blood was higher in the cell transplantation group than the cell control and healthy control groups（P〈0.05）; with time, the total number of leukocytes and neutrophils was decreased gradually.（3） Compared with the cell control and healthy control groups, the interleukin-10 level in the peripheral blood was lower and the levels of tumor necrosis factor-α and granulocyte colony-stimulating factor were higher at 1 day after cell transplantation（P〈0.05）. With time, in the cell transplantation group, the interleukin-10 level was increased gradually, the level of tumor necrosis factor-α was decreased gradually, and the level of granulocyte colony-stimulating factor was increased first and then decreased, which was highest at 7 days after cell transplantation.（4） Partial CM-Dil-positive cells were positive for CC16. Taken together, bone marrow mesenchymal stem cell transplantation via the tail vein can improve lung injury of rats with chronic obstructive pulmonary disease, and it is involved in the repair of airway injury through differentiation into epithelial cells and immune regulation.