增龄过程中人成骨细胞骨形成功能及相关基因改变研究

The underlying mechanisms of age-related osteopenia are still unknown. The purpose of the study is: ① to investigate the differences in morphology, proliferation, function and gene expression of cultured human ostoblastic cells in different age donors in vitro, in order to obtain the mechanism of senescence of osteoblasts. ② to compare the differences in bone formation and bone-related gene expressions between serially passaged cultures of osteoblasts from ≤5 year-old donor and osteoblasts from ≥70 year-old donors, in order to establish the model of osteoblastic senescence in vitro.The results showed that the aging osteoblasts were larger, thinner, containing more vacuoles and with elongated cell apophysis. Transmission electron microscope showed that the osteoblasts with donor aging became thin and elongated, with fewer cell organelles and undeveloped Golgi complex, with accumulated glycogen and more lysosomes in the plasma, and with more heterochromatin in the nuclear. No typical mineralizing nodule formation was observed in cultures from any age donors, but it could be found much tropocollagen in the extracellular matrix by transmission electron microscope when the osteoblasts were cultured with ascorbate and β-glycerophosphate. The cell number at confluence and cell proliferation ratio were shown to decrease with increasing donor age (P<0.01, respectively). ALP relative ratio was showen nonsignificant increase with increasing donor age. Quantity of total RNA in osteoblasts was showen to decreased with donor aging when the osteoblasts were cultured for 14 days (P<0.001 respectively). Expression of BGP, IGF-1, OPG, ODF, COL-1, TGF-β, ON and PTH/PTHrP-R mRNA could be detected in human osteoblasts of different age donors cultured in vitro. Quantity of BGP, IGF-1, OPG, OPG/ODF, COL-1 mRNA expression in the osteoblasts by semi-quantity RT-PCR analyses was showen to increase from ≤5 year-old donors to 30-40 year-old donors, then decreased with donor aging. Their expressions were correlated to donor age (P<0.05 or P<0.01 respectively). There is no significant difference between sexes. The morphology characteristics and proliferation of 10th-15th passage cells of infant donor were similar to that of osteoblasts from ≥70 year-old donors. mRNA expressions of BGP, IGF-1, OPG, OPG/ODF were decreased with increasing passages, which was similar to donor aging

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