CAJ | 학술논문

目的研究骨髓间充质干细胞( MSC)对博来霉素(BLM)所致大鼠肺间质纤维化治疗作用的可能机制.方法将54只雌性Wister大鼠按随机数字表法分为对照组、BLM组和MSC组,每组18只.对照组气管内注入生理盐水,BLM组气管内注入博来霉素,MSC组气管内注入博来霉素后,立即经尾静脉注入雄性大鼠MSC液0.5 ml(细胞数为2.5×106个),分别于第7、14、28天各组均处死6只大鼠.检测MSC移植前的5-溴-2-脱氧尿苷(BrdU)标记率,留取肺组织行病理学检查,测定羟脯氨酸含量,采用双重免疫荧光法检测BrdU标记的MSC是否表达Ⅱ型肺泡细胞(ATⅡ)特异性标志物肺表面活性蛋白-C(SP-C)；RT-PCR法检测不同时期肺组织和骨髓中SP-C mRNA表达量,观察肺损伤后自身骨髓动员是否参与Ⅱ型肺泡细胞的修复.结果 BrdU标记的MSC于48 h终浓度为10 μmol/L时标记率＞98％,且传代细胞可连续标记.MSC组7、14、28 d的肺组织冰冻切片中均可见到BrdU标记的MSC定位于肺组织并同时表达SP-C.28 d时MSC组与BLM组肺纤维化程度评分分别为(2.17±0.26)分和(2.83±0.24)分,肺组织羟脯氨酸含量分别为( 138 ±21)mg/g和(184±19)mg/g.肺组织中SP-C mRNA的表达量分别为0.98±0.15和0.59 ±0.14,且两组在不同时期骨髓中SP-C mRNA表达量均较对照组明显增加.结论骨髓间充质干细胞可定植于肺组织,并转化为Ⅱ型肺泡细胞,可阻止肺纤维化的进展,同时损伤诱导自身骨髓动员增强也参与修复过程.
목적 연구골수간충질간세포( MSC)대박래매소(BLM)소치대서폐간질섬유화치료작용적가능궤제.방법 장54지자성Wister대서안수궤수자표법분위대조조、BLM조화MSC조,매조18지.대조조기관내주입생리염수,BLM조기관내주입박래매소,MSC조기관내주입박래매소후,립즉경미정맥주입웅성대서MSC액0.5 ml(세포수위2.5×106개),분별우제7、14、28천각조균처사6지대서.검측MSC이식전적5-추-2-탈양뇨감(BrdU)표기솔,류취폐조직행병이학검사,측정간포안산함량,채용쌍중면역형광법검측BrdU표기적MSC시부표체Ⅱ형폐포세포(ATⅡ)특이성표지물폐표면활성단백-C(SP-C)；RT-PCR법검측불동시기폐조직화골수중SP-C mRNA표체량,관찰폐손상후자신골수동원시부삼여Ⅱ형폐포세포적수복.결과 BrdU표기적MSC우48 h종농도위10 μmol/L시표기솔＞98％,차전대세포가련속표기.MSC조7、14、28 d적폐조직빙동절편중균가견도BrdU표기적MSC정위우폐조직병동시표체SP-C.28 d시MSC조여BLM조폐섬유화정도평분분별위(2.17±0.26)분화(2.83±0.24)분,폐조직간포안산함량분별위( 138 ±21)mg/g화(184±19)mg/g.폐조직중SP-C mRNA적표체량분별위0.98±0.15화0.59 ±0.14,차량조재불동시기골수중SP-C mRNA표체량균교대조조명현증가.결론 골수간충질간세포가정식우폐조직,병전화위Ⅱ형폐포세포,가조지폐섬유화적진전,동시손상유도자신골수동원증강야삼여수복과정.
Objective To study the possible mechanisms of marrow mesenchymal stem cells (MSC) in therapy of bleomycin (BLM)-induced pulmonary fibrosis in rats.Methods Fifiy-four female Wistar rats were randomly divided into a control group,a BLM group and a MSC group.The control group received intratracheal normal saline,the BLM group received intratracheal instillation of bleomycin,and the MSC group was injected with male rat MSC solution of 0.5 ml (2.5 × 106 cells) via the tail vein after intratracheal instillation of bleomycin. Six rats from each group were killed on day 7,14 and 28 of the experiments.BrdU labeling rate was measured before MSC transplantation. Lung tissue specimens were obtained for pathological examination,hydroxyproline content measurement,and detection of the expression of type Ⅱ alveolar cell ( AT Ⅱ ) specific marker-pulmonary surfactant protein-C (SP-C) in BrdU labeled MSC using dual immunofluorescence method.RT-PCR method was used to detect SP-C mRNA expression in the lung tissue and the bone marrow at different stages.The bone marrow mobilization involved in repair of type Ⅱ alveolar cells after lung injury was observed.Results The final concentration of BrdU labeled MSC at 48 h was 10 μmoL/L, while the labeling efficiency was＞98％, and the passage cells could be continuously labeled.In the MSC group,BrdU labeled MSCs with expression of SP-C were observed in all frozen sections of lung tissue at day 7,14,and 28.By day 28,the lung fibrosis scores of the MSC group and the BLM group were (2.17 ± 0.26) and (2.83 ± 0.24),respectively,the lung tissue hydroxyproline contents were (138 ± 21 ) mg/g and (184 ± 19) mg/g,respectively,and the lung tissue SP-C mRNA expressions were (0.98 ± 0.15) and (0.59 ± 0.14),respectively. For both groups the SP-C mRNA expressions in the bone marrow at different stages were significantly increased as compared to the control group.Conclusions Marrow mesenchymal stem cells could be transplanted into lung tissues of rats,and transformed into type Ⅱ alveolar cells and was shown to prevent the development of pulmonary fibrosis.The damage-induced enhancement of host bone marrow mobilization was also involved in the repair process.