稀土化合物TbCl3对成骨细胞系MC3T3-E1增殖、分化和矿化功能的影响

Effects of TbCl_3 on the Proliferation, Osteogenic Differentiation and Mineralization Function of MC3T3-E1 Cells in Vitro

在细胞和分子水平上,研究了稀土化合物氯化铽(TbCl_3)对成骨细胞MC3T3-E1增殖、分化及矿化功能的影响。结果表明,细胞水平上,浓度为0.000 1、0.001、0.01、0.1、1和10μmol·L-1的TbCl_3均促进MC3T3-E1细胞的增殖、分化及其矿化功能,然而,当浓度升至为100和1 000μmol·L-1时,TbCl_3表现出抑制作用。分子水平上,浓度为0.000 1和0.1μmol·L-1的TbCl_3明显上调成骨分化相关基因骨形成蛋白2(BMP-2),碱性磷酸酶(ALP),骨涎蛋白(BSP),Ⅰ型胶原蛋白(ColⅠ),骨钙素(OCN)和runt相关转录因子2(Runx2)的表达。浓度为1 000μmol·L-1的TbCl_3则抑制上述成骨分化相关基因的表达。浓度为0.000 1、0.1和1μmol·L-1的TbCl_3促进成骨分化相关蛋白Runx2,BMP-2和OCN的表达;结果显示,低浓度的TbCl_3促进MC3T3-E1细胞的成骨分化及矿化功能,而高浓度TbCl_3则呈现出抑制作用。TbCl_3通过调控Runx2的表达刺激早期成骨分化相关基因BMP-2、ColⅠ和晚期成骨分化相关基因ALP、OCN的表达,从而诱导MC3T3-E1成骨分化。

The effects of TbCl3 on the proliferation, differentiation, and mineralization of a murine preosteoblast cell line MC3T3-E1 at cell and molecule levels were investigated in vitro. The results showed that TbCl3 with concentrations of 0.000 1, 0.001, 0.01, 0.1, 1, and 10 μmol·L-1promoted the proliferation,differentiation, and mineralization of MC3T3-E1 cells, while the TbCl3 with the concentrations of 100 and1 000 μmol·L-1showed the contrary effects. On the molecular level, the expressions of bone morphogenetic protein 2（BMP-2）, alkaline phosphatase（ALP）, bone sialopmtein（BSP）, Collagen Type Ⅰ（Col Ⅰ）,osteocalcin（OCN）, and runt-related transcription factor 2（Runx2） genes were up-regulated by 0.000 1 and0.1 μmol·L-1TbCl3 treatments through the reverse transcriptase polymerase chain reaction（RT-PCR）, but TbCl3 with the concentration of 1 000 μmol·L-1down-regulated the expressions above osteogenic differentiation related genes. The expressions of Runx2, BMP-2, and OCN proteins were up-regulated by 0.000 1,0.1 and 1 μmol·L-1TbCl3 treatments. The results suggested that TbCl3 with lower concentrations promoted,but higher concentrations inhibited the proliferation, osteogenic differentiation, and mineralization of MC3T3-E1 cells. TbCl3 likely regulated the expression of Runx2, which subsequently stimulated osteoblasts marker genes BMP-2 and Col Ⅰ at early stages and ALP and OCN at later stages of differentiation, thus promoted the proliferation, osteogenic differentiation and mineralization of MC3T3-E1 cells.