CAJ | 학술논문

目的：：探讨气管内移植人脐血间充质干细胞( HUCBMSCs ）对于博来霉素诱导的小鼠肺纤维化的作用，比较气管内及尾静脉移植HUCBMSCs对肺纤维化的治疗效果。方法：取第2代HUCBMSCs培养至4代6；0只无特定病原体的雄性昆明小鼠随机分成4组：阴性对照组( Cont组）、博来霉素组( BLM组）、HUCBMSCs移植Ⅰ组( MⅠ组）和HUCBMSCs移植Ⅱ组( MⅡ组），每组15只；后3组小鼠采用气管内插管灌注博来霉素制造肺纤维化模型2，4 h后，MⅠ组各小鼠气管内灌注5-溴-2-脱氧尿嘧啶核苷(Brdu）标记的HUCBMSCs，MⅡ组各小鼠尾静脉内注射Brdu标记的HUCBMSCs，分别于模型建造后的第7、14、28天每组各处死5只小鼠取材。采用HE染色和Masson染色观察肺组织形态学变化；免疫组织化学法检测间充质干细胞在肺组织的定位和分布；碱水解法测定羟脯氨酸的水平；免疫印迹法检测TGF-β1、α-SMA的蛋白表达量。结果：两组间充质干细胞移植组第7、14、28天均可见Brdu标记的细胞；3个时间点的MⅠ组、Ⅱ组的肺泡炎症和肺纤维化程度均分别较博来霉素组轻，差异有统计学意义(P＜0.05）。肺组织中TGF-β1、α-SMA的表达情况：BLM组＞MⅠ组、MⅡ组＞Cont组(P＜0.05）；MⅠ组和Ⅱ组表达量差异无统计学意义( P＞0.05）。结论：气管内移植HUCBMSCs可以定植于受损的肺组织中，能够减轻博来霉素诱导的小鼠肺纤维化。
목적：：탐토기관내이식인제혈간충질간세포( HUCBMSCs ）대우박래매소유도적소서폐섬유화적작용，비교기관내급미정맥이식HUCBMSCs대폐섬유화적치료효과。방법：취제2대HUCBMSCs배양지4대6；0지무특정병원체적웅성곤명소서수궤분성4조：음성대조조( Cont조）、박래매소조( BLM조）、HUCBMSCs이식Ⅰ조( MⅠ조）화HUCBMSCs이식Ⅱ조( MⅡ조），매조15지；후3조소서채용기관내삽관관주박래매소제조폐섬유화모형2，4 h후，MⅠ조각소서기관내관주5-추-2-탈양뇨밀정핵감(Brdu）표기적HUCBMSCs，MⅡ조각소서미정맥내주사Brdu표기적HUCBMSCs，분별우모형건조후적제7、14、28천매조각처사5지소서취재。채용HE염색화Masson염색관찰폐조직형태학변화；면역조직화학법검측간충질간세포재폐조직적정위화분포；감수해법측정간포안산적수평；면역인적법검측TGF-β1、α-SMA적단백표체량。결과：량조간충질간세포이식조제7、14、28천균가견Brdu표기적세포；3개시간점적MⅠ조、Ⅱ조적폐포염증화폐섬유화정도균분별교박래매소조경，차이유통계학의의(P＜0.05）。폐조직중TGF-β1、α-SMA적표체정황：BLM조＞MⅠ조、MⅡ조＞Cont조(P＜0.05）；MⅠ조화Ⅱ조표체량차이무통계학의의( P＞0.05）。결론：기관내이식HUCBMSCs가이정식우수손적폐조직중，능구감경박래매소유도적소서폐섬유화。
Objective:To explore the effect of intratracheal transplantation human umbilical cord blood mesenchymal stem cells on pulmonary interstitial fibrosis by bleomycin in mice,compare the treatment in pulmonary fibrosis of intratracheal transplantation with tail vain injection human umbilical cord blood mesenchymal stem cells.Methods:The second generation of HUCBMSCs were cultured to the fourth generation.Sixty specific pathogen free male Kunming mice were randomly divided into 4 groups:negative control group ( Cont group) ,bleomycin group( BLM group) ,HUCBMSCs transplantation groupⅠ( MⅠgroup) and HUCBMSCs transplantation groupⅡ(MⅡ group),each group 15 mice.Pulmonary fibrosis models were induced by bleomycin via intratracheal perfusion in the latter three groups.Twenty four hours after model establishment,5-bromo-2-deoxyuridine( Brdu) marked HUCBMSCs were poured in trachea in MⅠgroup ,the same were injected into tail vein in MⅡ group.At the 7th,14th,28th day,5 mice in each group were executed re-spectively.The morphological changes of the lung tissues were observed by HE staining and Masson staining.The localization and distribution of human umbilical cord blood mesenchymal stem cells were determined by the method of immunohistochemistry.The hydroxyproline contents were measured by alkali hydrolysis assay.The protein levels of transforming growth factor β-1 ( TGF-β1 ) and smooth muscle alpha-actin(α-SMA) were detected by Western blot.Results:In the two mesenchymal stem cell transplantation groups, there were Brdu marked cells at the 7th,14th,28th days in lung tissue.The alveolitis and fibrosis in lung of the two mesenchymal stem cell transplantation groups were milder than which of the the bleomycin group(P<0.05).Hydroxyproline levels in the bleomycin groups at 3 time points showed a gradually rising trend,the highest level was at 28th day( P<0.01).The protein levels of TGF-β1 andα-SMA in the lung tissues ranked as follows:BLM group>MⅠ,MⅡgroup>Cont group(P<0.05),the difference between MⅠgroup and MⅡgroup was not statistically significant(P>0.05).Conclusion:The colonization of human umbilical cord blood mesenchymal stem cells can be seen in the damaged tissue via intratracheal transplantation which can alleviate pulmonary fibrosis in mice caused by bleomycin.