髓内脂肪细胞对骨髓基质细胞成骨能力的影响

Effects of adipocytes on osteogenic potential of bone marrow stromal cell

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 effects of adipocyte in bone marrow cavity on osteogenic potential,and study the pathogenic mechanism of adipocyte differentiation from bone marrow stromal cells in osteoporosis.Methods Bone marrow stromal cells were inoculated in culture plate with the concentration of 105/ml and divided into 5 groups,which were added with adipocytes with concentrations of 0,103,104,105,106/ml respectively.The co-culture system of adipocyte and bone marrow stromal cell was established.After 4,8,12 days,the samples were detected for the activity of alkaline phophatase,the mRNA expression of type I collagen by in situ hybrization,the collagen synthesis ability with 3H-proline in corporation test.[WT5”HZ]Results After 12 days,with the increase of concentration of adipocytes,the activity of alkaline phophatase was significantly different between experimental groups and control (P< 0.05 or P< 0.01).The mRNA expression level of type I collagen in control group was significantly higher than in other groups,and collagen synthesis ability was decreased in the experimental group.Conclusion Adipocytes in bone marrow cavity can inhabite osteogenic potential of bone marrow stromal cells.It might play an important role in osteoporosis.