骨髓增生异常综合征患者骨髓间充质干细胞成骨分化功能的研究  

Osteogenic Differentiation Potential of Bone Marrow Mesenchymal Stem Cells in Patients with Myelodysplastic Syndromes

作　　者：费成明[1] 顾树程[1] 赵佑山[1] 郭娟[1] 李晓[1] 常春康[1] 

机构地区：[1]上海交通大学附属第六人民医院血液科,上海200233

出　　处：《中国实验血液学杂志》2015年第3期750-755,共6页Journal of Experimental Hematology

基　　金：国家自然科学基金(81170463)

摘　　要：目的:研究骨髓增生异常综合征(myelodysplastic syndromes,MDS)患者骨髓间充质干细胞(bone marrow mesenchymal stem cells,BM M SC)成骨分化功能及其在M DS发病机制中的作用。方法:分离培养M DS患者及正常人的BMMSC,并在体外诱导其向成骨细胞分化;荧光定量PCR法检测BMMSC成骨分化转录因子Runt相关转录因子2(RUNX2)和Osterix的mRNA表达水平,以及成骨诱导分化第7、14、21天成骨相关基因碱性磷酸酶(alkaline phosphatase,ALP)、骨涎蛋白(Bone sialoprotein,BSP)、骨桥素(Osteopontin,OPN)和骨钙素(osteocalcin,OCN)的mRNA表达情况;碱性磷酸酶活性检测试剂盒检测成骨诱导第3、7、10天ALP活性;茜素红染色观察诱导分化第21天钙结节形成情况。结果:低危MDS组BMMSC成骨分化转录因子RUNX2和Osterix mRNA表达水平较正常对照组均明显降低(P<0.05),成骨诱导分化第3天低危MDS组ALP活性水平明显低于正常对照组(P<0.05),成骨诱导分化第21天茜素红染色显示,低危MDS组钙结节含量也明显少于正常对照组(P<0.05)。在成骨诱导分化不同时间点成骨早期标志ALP、BSP和后期标志OPN、OCN在低危MDS组中的mRNA表达水平也显著降低(P<0.05),而高危MDS组在一系列成骨分化检测指标上与正常对照组之间没有统计学差异。结论:低危MDS组BM MSC成骨分化能力明显减弱,而高危M DS组相对正常。异常的成骨分化功能可能是造成低危组M DS骨髓支持造血能力减弱的重要原因。Objective：To mvestigate the osteogemc differentlatlon potenttal of bone marrow mesencnymal stem ceils （BMMSC） in patients with myelodysplastic syndromes （MDS） and to explore the role of BMMSC osteogenic differentiation in the pathogenesis of MDS. Methods .. BMMSC were isolated from bone marrow of patients with MDS and healthy donors, then expanded in vitro. The expression of transcription factor gene RUNX2, Osterix and osteogenic differentiation markers （ALP, BSP, OPN, OCN） were measured by real-time PCR, the alkaline phosphatase （ALP） activity was assessed at 3, 7, 10 days after osteogenic differentiation. Mineralization analysis was performed at day 21 of osteogenic induction. Results：The expression level of RUNX2 and Osterix were significantly decreased in BMMSC from lower-risk MDS patients compared with normal controls （ P 〈 0.05 ）. After osteogenic induction, low-risk MDS showed lower alkaline phosphatase activity at day 3 （P 〈 0.05 ）, less intense alizarin red S staining at day 21 （P 〈 0.05 ）, and lower gene expression of osteogenic differentiation markers （ P 〈 0. 05 ）, however, these expressions in higher-risk MDS were normal. Conclution： BMMSC from low-risk MDS have abnormalities in osteogenic differentiation,it may contribute to the ineffective hamatopoiesis of MDS.

关 键 词：骨髓增生异常综合征 骨髓间充质干细胞 成骨分化 

分 类 号：R551.3[医药卫生—血液循环系统疾病]