地塞米松与TGF β_1对骨髓基质细胞增殖、分化及代谢的影响  

Influence of dexamethasone and TGFβ_1 on proliferation, differentiation and metabolism of bone marrow stromal cells

作　　者：周广东[1] 王晓云[1] 刘德莉[1] 吴娟娟[1] 崔磊[1] 刘伟[1] 曹谊林[1] 

机构地区：[1]上海第二医科大学附属第九人民医院组织工程实验室,上海200011

出　　处：《中国实用美容整形外科杂志》2004年第1期51-55,共5页

基　　金：国家 8 63重大专项基金资助 (2 0 0 2AA2 0 50 2 1 );国家自然科学基金资助 (30 30 0 353)

摘　　要：目的　明确地塞米松与转化生长因子 β1对骨髓基质细胞增殖代谢及向软骨细胞分化的影响 ,为应用骨髓基质细胞构建组织工程化软骨提供技术参数。方法　抽取猪股骨骨髓 ,分离有核细胞 ,体外培养扩增并分别加入地塞米松 (40ng/ml)或地塞米松 (40ng/ml)与转化生长因子β1(10ng/ml)联合诱导。通过细胞计数及MTT法检测不同诱导因子对骨髓基质细胞增殖的影响 ,透射电镜观察不同诱导组细胞代谢功能的变化 ,免疫组化、原位杂交、RT -PCR及Northernblott等检测Ⅱ型胶原及蛋白聚糖表达量的差异。结果　加入地塞米松后细胞增殖能力受到轻度抑制 ,而同时加入地塞米松与转化生长因子 β1后 ,细胞生长明显受抑制。电镜下可见随诱导因子的增加 ,细胞变得肥大 ,细胞器丰富 ,功能活跃。Ⅱ型胶原及蛋白聚糖的表达在联合诱导组明显增强 ,且随诱导时间的延长 ,Ⅱ型胶原mRNA表达量也显著增加 (P <0 .0 1)。结论　地塞米松与转化生长因子β1能有效地诱导骨髓基质细胞表达软骨细胞特征性蛋白 ,但对细胞的增殖及代谢也产生明显的影响 ,在骨髓基质细胞构建组织工程化软骨时 。Objective To confirm influence of dexamethasone and transforming growth factor-beta1(TGF β 1) on proliferation, differentiation and metabolism of bone marrow stromal cells(BMSCs) and to provide parameters for constructing cartilage using BMSCs as seed cell. Methods Bone marrows were harvested from 3 porcine femurs, BMSCs were isolated and in vitro expanded and induced with dexamethasone (40?ng/ml) or with both dexamethasone (40?ng/ml) and transforming growth factor beta1(TGF β 1, 10?ng/ml). The influence of proliferation and metabolism on BMSCs caused by induced factors was observed by MTT test, cell counting and transmission electron microscope. Expressions of collagen Ⅱ and aggrecan were tested through immunohistochemistry, in situ hybridization, RT-PCR and Northern bloting. Results Stronger suppression of proliferation for BMSCs was caused by both dexamethasone and TGF β 1 than caused by dexamethasone alone. Hypertrophic cells with active function were observed in BMSCs induced with both dexamethasone and TGF β 1 with transmission electron microscope. Stronger expression of collagen Ⅱ and aggrecan were observed in BMSCs induced with both dexamethasone and TGF β 1 than in cells induced with dexamethasone alone, furthermore, a remarkable increase of collagen Ⅱ mRNA was observed in BMSCs induced with both dexamethasone and TGF β 1 with prolongation of induced time. Conclusion Chondrogenic phenotype of BMSCs can be better induced by both dexamethasone and TGF β 1, meanwhile, proliferation and metabolism of BMSCs can be influenced by induced factors. So we should take the relationship of proliferation and induction into account sufficiently when we construct engineered cartilage using BMSCs as seed cell.

关 键 词：地塞米松 TGFΒ1 骨髓基质细胞 细胞增殖 细胞分化 细胞代谢 软骨组织工程 

分 类 号：R329.28[医药卫生—人体解剖和组织胚胎学] R971.1[医药卫生—基础医学]