Elsevier

Biochemical Pharmacology

Volume 59, Issue 5, 1 March 2000, Pages 471-475
Biochemical Pharmacology

Molecular and Cellular Pharmacology
Stimulatory effect of daidzein in osteoblastic MC3T3-E1 cells

https://doi.org/10.1016/S0006-2952(99)00351-2Get rights and content

Abstract

Daidzein is a natural isoflavone found in Leguminosae. The effect of daidzein on osteoblastic MC3T3-E1 cells was investigated. Cells were cultured in a serum-free medium for 48 hr in the presence of daidzein (10−7–10−5 M). Daidzein (10−6 and 10−5 M) caused a significant elevation of protein content, alkaline phosphatase activity, and DNA content in cells; those increases were about 1.4-, 1.5-, and 2.0-fold, respectively. The ability of daidzein (10−5 M) to increase protein content, alkaline phosphatase activity, and DNA content in cells was prevented completely by the presence of cycloheximide (10−6 M), an inhibitor of protein synthesis, indicating that the isoflavone has a stimulatory effect on cellular protein synthesis. The anabolic effect of daidzein (10−6 or 10−5 M) in cells was not distinguishable from that of genistein (10−6 or 10−5 M). Meanwhile, cell protein showed an additive effect of 17β-estradiol and daidzein, but their effects on alkaline phosphatase were not additive. Moreover, the effect of daidzein (10−5 M) in elevating cellular protein content and alkaline phosphatase activity was inhibited completely by the presence of tamoxifen (10−6 M), suggesting that the effect of the isoflavone is mediated partly through estrogen action. This study demonstrates that daidzein has an anabolic effect in osteoblastic MC3T3-E1 cells.

Section snippets

Chemicals

α-Modification of Eagle’s Minimal Essential Medium was obtained from Flow Laboratories, Inc. FBS† 1 was obtained from Bioproducts Inc. BSA (fraction V), daidzein, genistein, tamoxifen, cycloheximide, and 17β-estradiol were purchased from the Sigma Chemical Co. All other chemicals were reagent grade and obtained from Wako Pure Chemical Industries, Ltd. Tissue culture plastic dishes were purchased from Falcon Plastics. Other materials used were commercial products of the highest grade available.

Cell culture

Effect of daidzein in osteoblastic cells

The effect of daidzein on protein content in osteoblastic MC3T3-E1 cells is shown in Fig. 1. Cells were cultured for 48 hr in a medium containing either vehicle (1% ethanol) or daidzein (10−7–10−5 M). The presence of 10−6 and 10−5 M daidzein caused a significant increase in protein content, whereas 10−7 M daidzein had no effect (Fig. 1A). When cells were cultured for 48 hr in a medium containing either vehicle, daidzein (10−5 M), cycloheximide (10−6 M), or cycloheximide (10−6 M) plus daidzein

Discussion

Nutritional and pharmacological factors are needed to prevent bone loss with aging [14]. Genistein and daidzein are natural isoflavones found in Leguminosae. Recently, it has been reported that genistein can inhibit ovariectomy-induced bone loss, suggesting a role for it in the prevention of osteoporosis 3, 4, 6. The effect of daidzein on bone metabolism, however, is poorly understood. More recently, it has been found that daidzein can stimulate bone formation and mineralization in bone tissue

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