人骨髓间充质干细胞体外成骨与成脂分化过程中FGF1及其受体表达的变化  

The expressions of FGF1 and its receptor during the osteogenic and adipogenic differentiation of human bone marrow stromal cells

作　　者：董平[1] 焦虎[1] 李秋晨[1] 吕晓岩[1] 尹艳花[1] 肖苒[1] 

机构地区：[1]中国医学科学院北京协和医学院整形外科医院研究中心,北京100144

出　　处：《基础医学与临床》2013年第8期941-946,共6页Basic and Clinical Medicine

基　　金：国家自然科学基金(30871433;31071305)

摘　　要：目的研究人骨髓间充质干细胞(hBMSCs)成骨与成脂分化过程中成纤维细胞生长因子1(FGF1)及其受体(FGFRs)的动态表达,探讨FGF1在hBMSCs成骨与成脂分化过程中的作用。方法密度梯度离心法分离hBMSCs,取第3代细胞分别进行成骨和成脂诱导分化并染色鉴定。通过real-time PCR方法检测成骨和成脂标志基因以及FGF1/FGFRs的表达,免疫荧光染色法检测诱导分化前后FGF1的表达。Real-time PCR及油红O染色鉴定外源性FGF1对hBMSCs成骨与成脂分化的影响。结果体外诱导hBMSCs成骨和成脂分化后茜素红和油红O染色结果呈阳性;FGF1和FGFRs在诱导过程中呈动态表达;免疫荧光染色结果表明FGF1主要在细胞质中分布,成骨诱导3 d后在细胞核中的表达增加;相比对照组,加入外源的FGF1组7 d后脂滴数量明显减少,成脂标志基因表达受到抑制(P<0.01),而加入外源的FGF1 3 d骨桥蛋白(OPN)表达升高(P<0.05)。结论 FGF1能够抑制hBMSCs的成脂分化,并且可能通过促进骨桥蛋白表达而提前hBMSCs的成骨矿化。Objective To study the dynamic expression profiles of fibroblast growth factor 1 （ FGF1 ） and its receptors （FGFRs） during osteogenic and adipogenic differentiation of human bone marrow stromal cells （hBMSCs）, and fur- ther exploring the role of FGF1 during hBMSCs differentiation. Methods Cells were isolated from human bone mar- row by density gradient centrifugation method, hBMSCs in the 3rd passage were induced into osteoblasts and adipo- cytes respectively, and followed by staining for identification. The expressions of FGF1, FGFRs, and marker genes during osteogenic and adipogenic differentiation, were detected by real-time PCR. Immunofluorescent staining was applied for FGF1 localization of hBMSCs after induced differentiation. Meanwhile, the influence of exogenous FGF1 on osteogenic and adipogenic differentiation was detected by real-time PCR and Oil Red O stainning. Results Osteo- genic and adipogenic induced hBMSCs were stained by Mizarin Red S and Oil Red O. The expression status of FGF1 and its receptors showed dynamic variation. Immunofluorescent staining results illustrated that FGF1 was mainly dis- tributed in cytoplasm, and its expression amount increased in nucleus after osteogenic induction for 3 days. The ex- pression of osteopontin increased 0. 05）, and the amount of lipid during adipogenic differentiation during osteogenic differentiation in the presence of exogenous FGF1 for 3 days （P 〈 droplet decreased and the expressions of adipogenic marker genes were suppressed after the introduction of exogenous FGF1 for 7 days （P 〈 0. 01 ）. Conclusions FGF1 may inhibit adipogenic differentiation of hBMSCs, and promotes the expression of OPN, which may further has- ten the maturity of osteogenesis.

关 键 词：骨髓间充质干细胞 成骨分化 成脂分化 FGF1 FGFRs 

分 类 号：R329[医药卫生—人体解剖和组织胚胎学]