Itoh Fumiaki, Aoyagi Shigemi, Furihata-Komatsu Hanako, Aoki Masato, Kusama Hiroshi, Kojima Masami, Kogo Hiroshi
Central Research Laboratory, R&D, Kissei Pharmaceutical Co Ltd, 4365-1, Kashiwabara, Hotaka, Minamiazumi, Nagano 399-8304, Japan.
Eur J Pharmacol. 2003 Sep 5;477(1):9-16. doi: 10.1016/j.ejphar.2003.08.011.
We investigated the direct effects of various bisphosphonates on osteoblasts. At 10(-5) M, clodronate increased alkaline phosphatase activity in cultured MC3T3-E1 (osteoblast-like line) and ST2 (pluripotent mesenchymal line) cells. Etidronate significantly increased alkaline phosphatase activity at 10(-5) M only in MC3T3-E1 cells. These effects were due to an increase in alkaline phosphatase-positive cell numbers, and the differentiation-enhanced cells were capable of mineralization (von Kossa stain). Other bisphosphonates (pamidronate, alendronate, and incadronate) did not increase alkaline phosphatase activity in either cell line. In cultured rat calvariae, clodronate stimulated the expression of genes for alkaline phosphatase and osteocalcin (osteoblast-differentiation markers), but decreased the expression of the gene for tartrate-resistant acid phosphatase (osteoclast marker). Clodronate, etidronate, and incadronate inhibited protein Tyr phosphatase and Ser/Thr phosphatase activities in MC3T3-E1 cells. These data suggest that clodronate acts directly on mesenchymal cells to enhance osteoblast differentiation, and this effect may be partly expressed through inhibition of protein Tyr phosphatase and/or Ser/Thr phosphatase activity.
我们研究了各种双膦酸盐对成骨细胞的直接作用。在10⁻⁵ M浓度下，氯膦酸盐增加了培养的MC3T3 - E1（成骨细胞样细胞系）和ST2（多能间充质细胞系）细胞中的碱性磷酸酶活性。依替膦酸盐仅在10⁻⁵ M时显著增加了MC3T3 - E1细胞中的碱性磷酸酶活性。这些作用归因于碱性磷酸酶阳性细胞数量的增加，且分化增强的细胞能够矿化（冯·科萨染色）。其他双膦酸盐（帕米膦酸盐、阿仑膦酸盐和因卡膦酸盐）在两种细胞系中均未增加碱性磷酸酶活性。在培养的大鼠颅骨中，氯膦酸盐刺激了碱性磷酸酶和骨钙素（成骨细胞分化标志物）基因的表达，但降低了抗酒石酸酸性磷酸酶（破骨细胞标志物）基因的表达。氯膦酸盐、依替膦酸盐和因卡膦酸盐抑制了MC3T3 - E1细胞中的蛋白酪氨酸磷酸酶和丝氨酸/苏氨酸磷酸酶活性。这些数据表明，氯膦酸盐直接作用于间充质细胞以增强成骨细胞分化，且这种作用可能部分通过抑制蛋白酪氨酸磷酸酶和/或丝氨酸/苏氨酸磷酸酶活性来表达。
