【摘要】 目的　探讨SB2 0 3 5 80和PD980 5 9对骨髓间质干细胞 (MSC)分化为脂肪细胞的影响。方法采用Ficoll Paque淋巴细胞分离液分离成人肋骨骨髓MSC ,体外扩增 ,流式细胞仪进行表面标志的检测。用地塞米松、3 异丁基 1 甲基黄嘌呤 (IBMX)、胰岛素、消炎痛等定向诱导MSC分化为脂肪细胞 ;分化诱导液中加入不同剂量的SB2 0 3 5 80、PD980 5 9,观察其对脂肪细胞形成的影响。结果　成人骨髓MSC在体外扩增 15代可获得 ( 2～ 3 )× 10 12 个细胞。流式细胞仪检测结果显示CD2 9、CD44、CD90 、CD10 5、CD166表达阳性 ,CD14 、CD3 4 、CD45、CD11a为阴性。诱导 48h后 ,细胞内有脂滴出现 ,随着时间延长 ,脂滴逐渐增加并融合为脂泡。细胞计数结果显示 85 %以上细胞转变为脂肪细胞。细胞由梭形转变为圆形或多角性。加入不同剂量SB2 0 3 5 80均可抑制骨髓MSC分化为脂肪细胞 ,但PD980 5 9对MSC分化为脂肪细胞无明显影响。结论初步证明丝裂原激活蛋白酶 p3 8的活化可能在骨髓MSC分化为脂肪细胞过程中起重要作用。更多还原

【Abstract】 Objective To investigate the effects of SB203580 and PD98059 on the differentiation of bone marrow mesenchymal stem cells (MSC) into adipocytes. Methods Bone marrow MSC were separated from human rib marrow by Ficoll-Pague lymphocyte separating medium and proliferated in culture medium. To detect the surface antigens, the labeled cells were analysed by flow cytometer. MSC were induced with dexamethasone, insulin, 3-isobutyl-1-methyl-xanthine (IBMI) and indomethacin into adipocytes. SB203580 or PD98059 were added into various induction media and their effects on MSC were observed. The cells were stained with Oil Red O. The number of adipocytes were counted by phase-contrast microscope. Results Bone marrow MSC proliferated as undifferentiated cells in culture for 15 passages and (2-3)×10 12 cells were obtained. The isolated cultured MSC comprised a single phenotypic population and displayed a fibroblast-like morphology and were uniformly positive for CD 29 , CD 44 , CD 90 , CD 105 , CD 166 and did not express CD 14 , CD 34 , CD 45 , CD 11a . After being induced with induction medium, lipid vacuoles in MSC were observed at 48 h, and 2 weeks later, over 85% MSC differentiated into adipocytes with accumlation of lipid vacuoles fusing into lipid drop, as stained by Oil red O. The shape of these cells changed from spindle shape to round or polygonal shape. SB203580 blocked the adipogenic differentiation and PD98059 did not have obvious effect. Conclusion Activation of mitogen-activated protein kinase p38 may play an important role in the differentiation of bone marrow MSC into adipocytes.更多还原