过表达hMMP-1基因BMSC移植降解大鼠肝纤维研究  

Transplantation of h MMP-1 gene-modified BMSC attenunates liver fibrosis in rats

作　　者：杜超[1] 魏晓龙[1] 曾维政[1] 郑淑梅[1] 蒋明德[1] 

机构地区：[1]成都军区总医院消化内科,成都610083

出　　处：《中华细胞与干细胞杂志（电子版）》2015年第1期21-26,共6页Chinese Journal of Cell and Stem Cell（Electronic Edition）

基　　金：四川省卫生厅基金资助项目(20110091);成都军区总医院院管课题基金资助项目(YG110-A)

摘　　要：目的观察过表达人基质金属蛋白酶-1(h MMP-1)的骨髓间充质干细胞(BMSC)体内移植,降解大鼠纤维化肝脏中胶原纤维的作用。方法 50﹪CCL4植物油溶液3 ml/kg皮下注射10周构建肝纤维化大鼠模型;h MMP-1/BMSC通过尾静脉注射移植到肝纤维化大鼠体内,根据实验设计模型大鼠分为4组:A组注射1×106 e GFP-h MMP-1/BMSC(1 ml,n=10);B组注射1×106 e GFP-BMSC(1 ml,n=10);C组注射1×106BMSC(1 ml,n=10);D组注射1 ml生理盐水(n=10);组间比较采用单因素方差分析,两两比较采用Tukey检验。2周后检测肝脏胶原纤维含量、肝组织病理情况。结果移植后2周肝组织中可见GFP表达;Western Blot检测到A组中MMP-1表达增高(0.45±0.07,P=0.013),定量检测MMP-1酶活性为2.67×10-3mmol/(g min);肝组织苏木精-伊红染色(HE)见平均视野下A组纤维化程度显著改善;MASSON胶原染色及肝组织羟脯氨酸含量检测A组胶原含量明显降低(P=0.002);肝纤维化指标检测提示A组中透明质酸酶(HA)、层粘连蛋白(LN)及Ⅲ型前胶原(PCⅢ)含量显著下降(P<0.05);α-平滑肌肌动蛋白(α-SMA)含量在A组中显著下降(P=0.008)。结论体内移植BMSC并补充外源h MMP-1因子能有效降解大鼠纤维化肝脏中胶原纤维。Objective To investigate whether transplantation of hMMP-1 gene- modified BMSC can attenunate rat liver fibrosis induced by carbon tetrachloride （CCL4）. Methods The rat liver fibrosis model was established by subcutaneous injection of 3 ml/kg mixed CCL4 and vegetable oil （1：1） for 10 weeks. All the rats were randomly divided into 4 groups as follows： transplantation of 1 × 106 eGFP-hMMP-1/BMSC in Group A （ 1 ml, n = 10） ; transplantation of 1.00 × 106 eGFP/BMSC in Group B（ 1 ml, n = 10）; transplantation of 1.00 ~ 106 BMSC in Group C （1 ml, n = 10）; transplantation of lml saline in Group D （n = 10）. After two weeks, animals were sacrificed. We tested the content of liver collagen fiber and pathological changes of liver. Results The GFP positive cells can be detected in the frozen section of liver tissue 2 weeks after transplantation. Western Blot showed high expressions of hMMP-1 in liver in Group A, and the hMMP-1 activity was 2.67 × 10-3mmol/ （g.min） in Group A; HE staining showed that the average amount of pseudolobules, degree of cell degeneration and adipose degeneration was lowest in Group A （P = 0.0024） ; MASSON staining and the detection of hydroxyproline indicated that there was lest collagen in Group A （P = 0.0106） ; a significant reduction in the hyaluronic acid, laminin, procollagen III and a-smooth muscle actin was also observed in Group A （P = 0.0084）. Conclusions Transplantation of BMSC expressing hMMP - 1 factor can effectively degrade collagen fibers in the fibrosis of liver in rats.

关 键 词：间充质干细胞 基质金属蛋白酶1 基因 肝硬化 

分 类 号：R575.2[医药卫生—消化系统]