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目的探讨在骨髓腔中,来源于骨髓基质细胞的脂肪细胞对骨髓基质细胞成骨能力的影响。方法将骨髓基质细胞以1×106个/ml浓度接种于24孔培养板中,共分为5组,分别加入0、103、104、105、106/ml浓度的脂肪细胞悬液,建立脂肪细胞和骨髓基质细胞共培养体系。共培养12d,每4d取细胞样本,检测细胞内碱性磷酸酶活性,利用原位杂交方法检测I型胶原mRNA表达,3H脯氨酸掺入实验检测骨髓基质细胞胶原合成能力。结果经过12d培养,不同时段的共培养体系中,随着脂肪细胞浓度上升,骨髓基质细胞内碱性磷酸酶相对活性下降,各实验组明显低于对照组(P<0.05或P<0.01);I型胶原mRNA表达减弱,对照组染色最深,实验组着色较淡;各实验组3H脯氨酸掺入实验min-1值均低于同期对照组(P<0.05或P<0.01)。结论髓内脂肪细胞干扰了骨髓基质细胞的成骨能力,可能与原发性骨质疏松症的发病有关。
Objective To investigate the effect of bone marrow stromal cells derived from bone marrow cells on osteogenic ability of bone marrow stromal cells in bone marrow cavity. Methods The bone marrow stromal cells were seeded into 24-well plates at the concentration of 1 × 106 cells / ml and divided into 5 groups. Adipose cells were respectively cultured in the concentrations of 0, 103, 104, 105 and 106 / ml, And bone marrow stromal cells co-culture system. Cultures were cultured for 12 days. Cell samples were collected every 4 days. The alkaline phosphatase (ACP) activity in cells was detected. The expression of type I collagen mRNA was detected by in situ hybridization. 3H-proline incorporation assay was used to detect collagen synthesis ability of bone marrow stromal cells. Results After 12 days culture, the relative activity of alkaline phosphatase in bone marrow stromal cells decreased with the increase of adipocyte concentration in the co - culture system at different time points, which was significantly lower in each experimental group than that in the control group (P <0.05 or P <0.01). The mRNA expression of type I collagen was weakened, the staining of the control group was the deepest, and the staining of experimental group was lighter. The min-1 values of 3H-proline incorporation in each experimental group were lower than those in the same period (P <0.05 or P <0.01). Conclusion Intramedullary adipocytes interfere with the osteogenic capacity of bone marrow stromal cells, which may be related to the pathogenesis of primary osteoporosis.