摘要
目的:观察不同浓度地塞米松对大鼠骨髓基质细胞成脂、成骨分化能力的影响。方法:实验于2003-10/2005-01在兰州大学医学院中心实验室完成。①选取清洁级雌性6周龄Wistar大鼠6只,采用全骨髓法分离大鼠双侧股骨骨髓基质细胞,传至第3代细胞用于实验,随机分为4组:地塞米松成脂诱导组、单纯成脂诱导组、地塞米松成骨诱导组、单纯成骨诱导组。②传代后培养液中分别加成脂、成骨诱导剂,地塞米松成脂诱导组、地塞米松成骨诱导组培养基中均含有10-10,10-9,10-8,10-7,10-6,10-5mol/L不同浓度地塞米松,单纯成脂诱导组、单纯成骨诱导组培养基中无地塞米松,其余条件相同进行培养。通过细胞内三酰甘油含量和碱性磷酸酶活性的测定来比较不同浓度地塞米松在大鼠骨髓基质细胞分化中的作用。结果:不同浓度地塞米松对细胞内三酰甘油含量及碱性磷酸酶活性的影响:①培养21d,地塞米松成脂诱导组随地塞米松浓度的增加,细胞内三酰甘油含量明显增加,均高于单纯成脂诱导组(P<0.05或P<0.01)。②培养12d,地塞米松成骨诱导组地塞米松浓度为10-8mol/L时碱性磷酸酶活性增至最高,之后随地塞米松浓度的增高,碱性磷酸酶活性逐渐降低。结论:10-8mol/L以上高浓度地塞米松可诱导骨髓基质细胞分化为脂肪细胞,同时抑制其向成骨细胞的分化,可能与皮质类固醇激素性骨质疏松的发病有关。
AIM： To＇observe the effect of different concentrations of dexamethasone on differentiation of bone marrow stromal cell into adipocytes and osteoblasts.
METHODS： The experiment was carried out at Central Laboratory, Medical College, Lanzhou University, from October 2003 to January 2005. ①Bone marrow stromal cells（BMSCs）of six female SD rats, of clean grade, aged 6 weeks, were isolated from both femora in Wistar rats by using allbone marrow culture method. Cells of the third generation were used in the experiment and randomly divided into 4 groups： dexamethasone adipogenic induced group, simple adipogenic induced group, dexamethasone osteogenic induced group and simple osteogenic induced group. ②After subculture of the cells, the cells were induced to adipocytes or osteoblasts by adipogenic inducer or osteogenic inducer. Different concentrations（10^-10, 10^-9, 10^-8, 10-7,10^-6, 10^-5 mol/L） of dexamethasone were added into culture medium of dexamethasone adipogenic induced group and dexamethasone osteogenic induced group, and simple adipogenic induced group and simple osteogenic induced group were cultured without dexamethasone. The effect of dexamethasone on differentiation of BMSCs into adipocytes or osteoblasts was evaluated in vitro by measuring the contents of tfiglyceride and alkaline phosphatase activity.
RESULTS： Effect of different concentrations of dexamethasone on the content of triglyceride and the activity of alkaline phosphatase of BMSC：① After culture for 21 days, triglyceride content of dexamethasone adipogenic induced group were enhanced significantly, and higher than that of simple adipogenic induced group with the increase of concentration of dexamethasone （P〈0.05 or P〈0.01）. ②After culture for 12 days, the activity of alkaline phosphatase of dexamethasone osteogenic induced group was the highest when the concentration of dexamethasone was 10^-8 mol/L. But the activity of alkaline phosphatase was gradually declined when the concentration of dexamethasone was over 10^-8 mol/L. CONCLUSION： Dexamethasone could induce BMSC differentiating into adipocytes and inhibit osteoblastic differentiation when its concentration is over 10^-8 mol/L, which may be related to the pathogenetic mechanism of steroid-iriduced osteoporosis.
出处
《中国临床康复》
CSCD
北大核心
2006年第29期98-100,共3页
Chinese Journal of Clinical Rehabilitation
