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Inhibitory effect of menaquinone-7 (vitamin K2) on osteoclast-like cell formation and osteoclastic bone resorption in rat bone tissues in vitro

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Abstract

The effect of menaquinone-7 (MK-7; vitamin K2) on oateoclast-like cell formation and osteoclastic bone resorption in rat femoral tissues in vitro was investigated. The bone marrow cells were cultured for 7 days in a α-minimal essential medium (α-MEM) containing a well-known bone resorbing agent [parathyroid hormone (1–34) (PTH) or prostaglandin E2 (PGE2)] with an effective concentration. Osteoclast-like cells were estimated by staining for tartrate-resistant acid phosphatase (TRACP), a marker enzyme of osteoclasts. The presence of PTH (10–8 M) or PGE2 (10–6 M) induced a remarkable increase in osteoclast-like multinucleated cells. These increases were significantly inhibited by MK-7 (10–8–10–5 M). MK-7 (10–7 and 10–6 M) significantly inhibited phorbol 12-myristate 13-acetate-induced osteoclast-like cell formation, whereas MK-7 did not inhibit dibutyryl cyclic adenosine monophosphate (DcAMP) (10–5 M)-induced osteoclast-like cell formation. These results suggest that the inhibitory action of MK-7 is partly involved in protein kinase C signaling. The bone cells isolated from rat femoral tissues were cultured for 48 h in an α-MEM containing either vehicle or MK-7 (10–8–10–5 M). The presence of MK-7 (10–6 and 10–5 M) caused a significant decrease in the number of mature osteoclasts. Such a decrease was also seen in the presence of calcitonin (10–10–10–8 M), DcAMP (10–6 and 10–5 M), or calcium chloride (10–3 and 10–3 M). The effect of MK-7 (10–6 M) in decreasing the number of osteoclasts was not further enhanced in the presence of calcitonin (10–8 M), DcAMP (10–5M), or calcium chloride (10–3 M), and was completely abolished by the presence of dibucaine (10–6 M) or staurosporine (10–7 M), which are inhibitors of Ca2+-dependent protein kinases. These results suggested that MK-7 has a suppressive effect on osteoclasts. Moreover, the femoral-metaphyseal tissues obtained from rats were cultured for 48 h in Dulbecco's modified Eagle's medium containing either vehicle, PTH (10–7 M), or PGE2 (10–5 M) in the absence or presence of MK-7 (10–7–10–5 M). The presence of PTH or PGE2 induced a significant decrease in bone calcium content. These decreases were significantly blocked by MK-7 (10–7–10–5 M). This study demonstrates that MK-7 has an inhibitory effect on osteoclastic bone resorption in vitro.

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  1. Laboratory of Endocrinology and Molecular Metabolism, Graduate School of Nutritional Sciences, University of Shizuoka, 52-1, Yada, Shizuoka, 422-8526, Japan

    Masayoshi Yamaguchi & Zhong Jie Ma

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  1. Masayoshi Yamaguchi
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  2. Zhong Jie Ma
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Correspondence to Masayoshi Yamaguchi.

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Yamaguchi, M., Ma, Z.J. Inhibitory effect of menaquinone-7 (vitamin K2) on osteoclast-like cell formation and osteoclastic bone resorption in rat bone tissues in vitro. Mol Cell Biochem 228, 39–47 (2001). https://doi.org/10.1023/A:1013360308946

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