Department of Orthopaedics, Tonglu County Chinese Medicine Hospital, Hangzhou, China.
Department of Orthopaedics, Hangzhou Traditional Chinese Medicine Hospital, Hangzhou, China.
J Biol Regul Homeost Agents. 2018 May-Jun;32(3):497-505.
The aim of this work was to study the mechanisms of vitamin D receptor (VDR) and Genistein (Gen) on the regulation of bone metabolism of phytoestrogens from cellular and epidemiological perspectives. MC3T3-E1 cells were treated with different concentrations of Gen, and the cell-proliferation rate was detected by an MTT colorimetric assay. The effect of the VDR receptor blocker ZK159222 on the Gen effect was then observed; after adding Gen to MC3T3-E1 cells, we detected the expression of VDR protein via Western blotting. After adding estrogen receptor α-blocker MPP and estrogen receptor β-blocker PHTPP, we observed the effect of Gen on the regulation of the VDR protein. DNA was extracted from the blood samples of 200 postmenopausal women in the early epidemiological survey, and the restriction fragment length polymorphism of VDR gene Apa I and Bsm I in each sample was observed. The results were analyzed using dietary survey and bone mineral density examination. The results show that 10-8mol/L Gen can promote the proliferation of MC3T3-E1 cells (P less than 0.05). This effect can be canceled by the VDR blocker ZK159222. By analyzing the Apa I and Bsm I genotypes of VDR restriction sites, we discovered no significant difference in bone mineral density (BMD) between different genotypes (P>0.05). In addition, there was no significant correlation between dietary phytoestrogen intake and BMD in different genotypes (P>0.05). In conclusion, VDR can mediate the effect of Gen on the proliferation of MC3T3-E1 cells. The up-regulated expression of VDR protein in Gen is not mediated by the estrogen receptor. Moreover, the VDR gene polymorphism is not related to the BMD in various parts and is not related to the bone metabolism effect of the dietary plant estrogen intake.
本研究旨在从细胞和流行病学角度探讨维生素 D 受体（VDR）和金雀异黄素（Gen）对植物雌激素调节骨代谢的作用机制。用不同浓度的 Gen 处理 MC3T3-E1 细胞，采用 MTT 比色法检测细胞增殖率。观察 VDR 受体阻断剂 ZK159222 对 Gen 作用的影响；向 MC3T3-E1 细胞中加入 Gen 后，通过 Western blot 检测 VDR 蛋白的表达。加入雌激素受体 α 阻断剂 MPP 和雌激素受体 β 阻断剂 PHTPP 后，观察 Gen 对 VDR 蛋白调节作用的影响。从 200 例早期流行病学调查的绝经后妇女的血液样本中提取 DNA，观察每个样本中 VDR 基因 Apa I 和 Bsm I 的限制性片段长度多态性。利用膳食调查和骨密度检查对结果进行分析。结果表明，10-8mol/L Gen 可促进 MC3T3-E1 细胞的增殖（P<0.05），该作用可被 VDR 阻断剂 ZK159222 取消。通过分析 VDR 限制酶切位点的 Apa I 和 Bsm I 基因型，我们发现不同基因型之间的骨密度（BMD）无显著差异（P>0.05）。此外，不同基因型之间的膳食植物雌激素摄入量与 BMD 无显著相关性（P>0.05）。综上所述，VDR 可介导 Gen 对 MC3T3-E1 细胞增殖的作用，Gen 上调 VDR 蛋白的表达不通过雌激素受体介导。此外，VDR 基因多态性与各部位 BMD 无关，与膳食植物雌激素摄入量的骨代谢作用无关。
