胰岛素样生长因子Ⅰ转染对脂肪干细胞TWEAK/Fn14信号通路的影响  

Effect of adipose-derived stem cells modified by insulin-like growth factor I gene on TWEAK/Fn14 signaling pathway

作　　者：刘玉平[1] 王明明[1] 刘涛[1] 李明[1] 俞光荣[2] 

机构地区：[1]滕州市中心人民医院创伤骨科,山东省滕州市277500 [2]同济大学附属同济医院足踝外科,上海市200065

出　　处：《中国组织工程研究》2015年第45期7217-7223,共7页Chinese Journal of Tissue Engineering Research

摘　　要：背景:胰岛素样生长因子Ⅰ具有诱导间充质干细胞向软骨细胞分化的能力,通过基因转染的方式将胰岛素生长因子Ⅰ转染入脂肪干细胞或许能够更好的促进脂肪干细胞向软骨细胞分化。目的:探讨胰岛素样生长因子Ⅰ基因对脂肪间充质干细胞体外定向成软骨分化能力以及对TWEAK/Fn14信号通路的影响。方法:构建慢病毒载体p LVX-IGF-I-IRES-Zs Greenl基因并转染第3代人脂肪间充质干细胞,诱导细胞向软骨分化。同时将pL VX-IRES-Zs Greenl转染的脂肪间充质干细胞设为绿色荧光蛋白/脂肪间充质干细胞组,单纯脂肪间充质干细胞设为对照组。结果与结论:与绿色荧光蛋白/脂肪间充质干细胞组和对照组相比,胰岛素样生长因子Ⅰ/脂肪间充质干细胞组细胞中TWEAK mR NA表达水平降低,而胰岛素样生长因子Ⅰ,Col2al和Sox9 mR NA的表达水平增加,Col2a1表达水平增加,基质金属蛋白酶3以及TWEAK的表达降低。提示pL VX-IGF-I-IRES-ZsG reenl慢病毒转染脂肪间充质干细胞后可获得胰岛素样生长因子Ⅰ的高效表达,且能下调细胞TWEAK基因及蛋白表达,可促进体外培养的脂肪间充质干细胞转化为软骨细胞。BACKGROUND： Because insulin-like growth factor I has the ability to induce mesenchymal stem cells into chondrocytes, we hypothesized that the chondrogenic differentiation of adipose-derived stem cells can be improved via insulin-like growth factor I transfection.OBJECTIVE： To investigate the influence of insulin-like growth factor I transfection on chondrogenic potential of adipose-derived stem cells in vitro and tumor necrosis factor-like weak inducer of apoptosis（TWEAK）/fibroblast growth factor-inducible 14（Fn14） signaling pathway. METHODS： Recombinant lentivirus plasmid pLVX-IGF-I-IRES-ZsGreenl was constructed and transferred into passage 3 adipose-derived stem cells, and then these stem cells were induced to differentiate into chondrocytes（experimental group 1）. Meanwhile, the cells transfected with pLVX-IRES-ZsGreenl were taken as green fluorescent protein/adipose mesenchymal stem cell group（experimental group 2）, and those with no transfection acted as control group. RESULTS AND CONCLUSION： The mRNA expression of TWEAK was reduced in the experimental group 1 as compared with the other two groups, but the mRNA expressions of insulin-like growth factor I, Col2a1 and Sox9 were up-regulated in the experimental group 1. At the same time, the protein expression of matrix metalloproteinase-3 and TWEAK were down-regulated, while the protein expression of Col2a1 was increased inthe insulin-like growth factor I-transfected cells in contrast to the cells modified with pLVX-IRES-ZsGreenl or with no transfection. These findings indicate that pLVX-IGF-I-IRES-ZsGreenl transfection of adipose-derived stem cells results in a higher expression of insulin-like growth factor I, and down-regulates the expression of TWEAK mRNA and protein, which improves the differentiation of adipose-derived mesenchymal stem cells into chondrocytes.

关 键 词：软骨细胞 胰岛素样生长因子Ⅰ 转染 组织工程 干细胞 脂肪干细胞 脂肪间充质干细胞 基因转染 TWEAK Fn14 细胞培养 

分 类 号：R394.2[医药卫生—医学遗传学]