脐带间充质干细胞对博来霉素诱导小鼠肺纤维化的干预机制初探  

Therapeutic effects and the underlying mechanism of umbilical cord-derived mesenchymal stem cells for bleomycin induced lung injury in rats

作　　者：李玉坤[1] 王华[2] 姜纯国[3] 黄慧[3] 刘佳[3] 王艳勋[3] 肖凤君[2] 杨国儒 吴祖泽[2] 

机构地区：[1]山东省德州市人民医院呼吸内科 [2]军事医学科学院放射与辐射医学研究所 [3]中国医学科学院北京协和医院呼吸内科,100730 [4]山东省潍坊市第二人民医院呼吸与危重症医学科

出　　处：《中华结核和呼吸杂志》2013年第11期808-813,共6页Chinese Journal of Tuberculosis and Respiratory Diseases

基　　金：国家自然科学基金（81170055）

摘　　要：目的 观察脐带间充质干细胞（UC-MSC）对博来霉素诱导的小鼠肺纤维化模型的干预作用.方法 体外分离、培养新生儿UC-MSC.将88只6～8周C57BL/6雄性小鼠按随机数字表法分为4组,对照组（20只）气管内注入磷酸盐缓冲液（PBS）,3d后尾静脉注射生理盐水；干细胞组（20只）气管内注入PBS,3d后尾静脉注射UC-MSC；博来霉素组（24只）气管内注入博来霉素,3d后尾静脉注射生理盐水；博来霉素＋干细胞组（24只）气管内注入博来霉素,3d后尾静脉注射UC-MSC,于实验第21天处死各组小鼠,取小鼠肺组织,行HE、Masson和免疫组织化学染色检测基质金属蛋白酶（MMP）-2和金属蛋白酶组织抑制剂-1（TIMP-1）的表达.结果 （1）肺组织病理：博来霉素＋干细胞组肺泡炎、纤维化程度及胶原蛋白沉积较博来霉素组明显减轻（分别为1.55 ±0.51和2.16 ±0.77;1.45±0.60和2.32±0.82,均P＜0.05）,对照组和干细胞组差异无统计学意义（分别为0.35 ±0.49和0.37 ±0.50;0.45 ±0.69和0.32±0.58,均P＞0.05）.（2） MMP-2表达含量：博来霉素＋干细胞组MMP-2的表达明显低于博来霉素组（分别为1.59 ±0.59和2.37 ±0.68,P＜0.05）,对照组和干细胞组差异无统计学意义（分别为0.80±0.69和0.84±0.77,P＞0.05）.（3） TIMP-1表达含量：博来霉素＋干细胞组TIMP-1的表达明显高于博来霉素组（分别为1.95 ±0.58和0.79 ±0.71,P＜0.05）,对照组和干细胞组差异无统计学意义（分别为1.10±0.72和1.32±0.58,P＞0.05）.结论 UC-MSC植入可以减轻博来霉素导致的小鼠肺纤维化程度,可能是通过影响MMP-2和TIMP-1的表达而发挥作用；UC-MSC对正常肺组织无影响.Objective To study the efficacy of umbilical cord-derived mesenchymal stem cells （UC-MSCs） for bleomycin-induced pulmonary fibrosis in mice.Methods UC-MSCs were isolated from the umbilical cord after parental consent.One hundred C57BL/6 mice were randomly divided into 4 groups （12 of these for preliminary experiment）.Mice in the control group （n =20） were instilled with PBS via trachea and NS was injected via the tail vein after 3 days.Mice in the stem cell group （n =20） were instilled with PBS via trachea and were injected with MSC via the tail vein after 3 days.Mice in the bleomycin group （n =24） were instilled with bleomycin via trachea and NS was injected via the tail vein after 3 days.Mice in the bleomycin plus stem cell group （n =24） were instilled with bleomycin via trachea and were injected with MSCs via the tail vein after 3 days.All of the mice were sacrificed at the 21th day,and the lungs were immediately fixed with 4% paraformaldehyde for 48 h,embedded in paraffin and sectioned at 5 μmol/Lthickness.The sections were stained with hematoxylin and eosin （H＆E） and Masson-trichrome.Histopathological scoring of pulmonary fibrosis was performed according to Ashcroft＇s method.The concentrations of matrix metalloproteinases-2 and tissue inhibitor of metalloproteinase-lwere determined using immunohistochemistry.Results Compared with the bleomycin group,MSC transplantation significantly reduced pulmonary inflammation,fibrosis and deposition of collagen in the bleomycin plus stem cell group [（1.55±0.51） vs （2.16±0.77）,and （1.45±0.60） vs （2.32±0.82）,respectively,P〈0.05].There was no difference between the control group and the stem cell group [（0.35 ± 0.49） vs （0.37 ±0.50）,P 〉 0.05].The expression of MMP-2 in the bleomycin plus stem cell group was lower than the bleomycin group [（1.59 ± 0.59） vs （2.37 ± 0.68）,P 〈 0.05],but there was no difference between the control group and the stem cell group [（0.80 ± 0.69） vs （0.84 ± 0.77）

关 键 词：肺纤维化 博来霉素 间质干细胞 基质金属蛋白酶类 金属蛋白酶1组织抑制剂 

分 类 号：R563.9[医药卫生—呼吸系统]