脐带间充质干细胞联合盐酸多西环素对肺纤维化小鼠的早期干预效果  

Effect of umbilical cord-derived mesenchymal stem cell transplantation combined with doxycycline on mice with early stage pulmonary fibrosis

作　　者：杨宇[1] 袁晓梅[1] 赵欣[1] 吴敏娜[2] 高新愿[1] 许芝山 钟根深 郭悦鹏[1] 

机构地区：[1]新乡医学院第一附属医院呼吸科,卫辉453100 [2]新乡医学院基础医学院微生物教研室,新乡453000 [3]河南省神经病学研究所生物治疗实验室,卫辉453100

出　　处：《郑州大学学报（医学版）》2016年第4期489-494,共6页Journal of Zhengzhou University(Medical Sciences)

基　　金：国家自然科学基金资助项目81201765;新乡医学院研究生科研创新支持计划资助项目YJSCX20406Z

摘　　要：目的:探讨脐带间充质干细胞(UC-MSCs)联合盐酸多西环素对肺纤维化小鼠的早期治疗效果及作用机制。方法:健康成年清洁级雌性C57BL/6小鼠40只,采用随机数字表法分为5组(n=8):对照组(气管内注入生理盐水,48 h后尾静脉注射生理盐水)、模型组(气管内注入博来霉素,48 h后尾静脉注射生理盐水)、盐酸多西环素组(气管内注入博来霉素,48 h后腹腔注射盐酸多西环素,隔日注射1次,连用4次)、干细胞组(气管内注入博来霉素,48 h后尾静脉注射UC-MSCs)、联合组(气管内注射博来霉素,48 h后尾静脉注射UC-MSCs联合腹腔注射盐酸多西环素,隔日注射1次,连用4次)。于造模第28天,取小鼠右肺组织行HE、Masson染色,进行肺泡炎评分及肺纤维化评分;取左肺组织测定羟脯氨酸含量。采用免疫组化法检测MMP-9、TIMP-1蛋白的表达。结果:与模型组相比,干细胞组和盐酸多西环素组肺泡炎及肺纤维化程度减轻,羟脯氨酸含量降低,MMP-9蛋白的表达减弱,TIMP-1蛋白的表达升高,联合组以上指标改善最为显著(P<0.05)。结论:早期应用UC-MSCs联合盐酸多西环素能够显著减轻小鼠肺纤维化程度,其机制可能与提高TIMP-1表达、抑制MMP-9的表达有关。Aim： To investigate the effect of umbilical cord-derived mesenchymal stem cells（ UC-MSCs） transplantation and doxycycline in a mice model of pulmonary fibrosis and its mechanism. Methods： A total of 40 C57 BL /6 female mice were randomly allocated into 5 groups. Mice in control group were intratracheally injected with NS under operating microscope. Mice in other groups were intratracheally injected with bleomycin. After 48 h,mice in the control and model group were injected with normal saline via the tail vein,mice in the doxycycline group were injected with doxycycline via abdomen,mice in the stem cell group were injected with UC-MSCs via the tail vein,and mice in the combination treatment group were given UC-MSCs plus doxycycline. On the 28 th day,the lung tissue was collected. Histopathological scoring of alveolitis and pulmonary fibrosis was performed according to Szapiel＇s method. The content of hydroxyproline was determined using alkaline hydrolysis method. The expressions of MMP-9 and TIMP-1 were determined using immunohistochemistry.Results： Compared with the model group,the alveolitis and pulmonary fibrosis in the stem cell group and doxycycline group alleviated,the content of hydroxyproline and the expression of MMP-9 decreased,while the expression of TIMP-1 increased,and the above parameters improved most significantly in the combination treatment group（ P 0. 05）. Conclusion： UC-MSCs transplantation and doxycycline can relieve bleomycin-induced pulmonary fibrosis in mice. The mechanism might be related with inhibiting MMP-9 and inducing TIMP-1 expression.

关 键 词：脐带间充质干细胞 盐酸多西环素 肺纤维化 MMP-9 TIMP-1 小鼠 

分 类 号：R563[医药卫生—呼吸系统]