骨髓间充质干细胞抑制大鼠肺纤维化的作用机制  

Inhibitory Effect of Mesenchymal Stem Cells on Pulmonary Fibrosis in Rats

作　　者：王贤[1] 曾玉兰[1] 彭红星[1] 杨荣时[1] 

机构地区：[1]华中科技大学同济医学院附属梨园医院呼吸内科,武汉430077

出　　处：《华中科技大学学报（医学版）》2014年第3期300-303,共4页Acta Medicinae Universitatis Scientiae et Technologiae Huazhong

基　　金：湖北省自然科学基金资助项目(No.2010CHB01100)

摘　　要：目的观察骨髓间充质干细胞(MSCs)对大鼠肺纤维化抑制作用的机制。方法体外分离、培养、纯化4周龄SD大鼠的骨髓MSCs。SD实验大鼠随机分为4组(每组12只):正常对照组(气管内注入生理盐水)、模型组(气管内注入博莱霉素)、MSCs治疗早期组(造模后立即给予尾静脉注射MSCs)、MSCs治疗晚期组(造模后14d给予尾静脉注射MSCs),气管内注入博莱霉素用量为5mg/kg,正常对照组注入等体积生理盐水,尾静脉注射MSCs用量为1.0×106/mL DMEM培养液1mL。于第28天统一处死大鼠后取肺组织,肺组织病理切片行苏木精-伊红(HE)染色及Masson染色观察肺部炎症和纤维化情况,Western blot法检测肺组织转化生长因子-β1(TGF-β1)、基质金属蛋白酶-2(MMP-2)、基质金属蛋白酶组织抑制剂-1(TIMP-1)蛋白表达量。结果①成功分离培养MSCs并鉴定。②模型组肺泡炎和肺纤维化程度较正常对照组明显加重,MSCs治疗早期组肺泡炎和肺纤维化程度较模型组显著减轻,MSCs治疗晚期组肺泡炎和肺纤维化程度与模型组比较差异无统计学意义。③模型组TGF-β1、TIMP-1蛋白表达较正常对照组显著增加,MSCs治疗组TGF-β1、TIMP-1蛋白表达较模型组显著减少,MMP-2蛋白表达在各组大鼠间差异无统计学意义。结论骨髓MSCs可抑制博莱霉素诱导的肺纤维化,并且在肺损伤早期给予MSCs干预的疗效更好,其机制可能为降低TGF-β1蛋白的表达,调节MMP-2与TIMP-1之间的平衡。Objective To investigate the inhibitory effect of mesenchymal stem cells （MSCs） on pulmonary fibrosis in rats, Methods The bone mesenchymal stem cells （BMSCs） of 4-week-old SD rat were isolated, cultured and purified in vitro. Forty-eight SD rats were randomly divided into four groups （n= 12 each） normal control group （intratracheal injection of normal saline） ;model group （intratracheal injection of 5 mg/kg bleomycin） ;early MSCs group （injection of 1 × 10^6/mL MSCs via the caudal vein immediately after the model establishment） ;later MSCs group （injection of 1 × 10^6/mL MSCs via the caudal vein 14 days after the model establishment）. All the rats were sacrificed at 28 days. The pathological changes of lung tissues were observed by HE and Masson staining. The expressions of TGF-β1, MMP-2 and TIMP-1 were detected by Western blot. Results @BMSCs were successfully cultured and identified. （2）Pulmonary alveolitis and fibrosis were remarkably aggrava- ted in the model group compared with the normal control group. They were significantly alleviated in the early MSCs group compared with the model group. There was no significant difference in the degree of pulmonary alveolitis and fibrosis between the later MSCs group and model group. ③The expressions of TGF-β1 and TIMP-1 were significantly increased in the model group compared with those in the normal control group. They were significantly decreased in MSCs groups when compared with the model groups. No significant difference in the expression of MMP-2 was noted among groups. Conclusion BMSCs can effectively prevent the development of bleomycin-induced pulmonary fibrosis. The effect is even better when MSCs are administered in the lung injury at early stage,which may be related to the reduction of TGF-β1 and the regulation of MMP-2 and TIMP- 1 balance.

关 键 词：肺纤维化 间充质干细胞 转化生长因子-Β 基质金属蛋白酶 基质金属蛋白酶组织抑制剂 

分 类 号：R563.4[医药卫生—呼吸系统]