淫羊藿黄酮上调去卵巢大鼠骨髓基质细胞成骨分化和下调成脂分化过程中 DKK1 蛋白的动态表达。

Dynamic expression of DKK1 protein in the process whereby Epimedium-derived flavonoids up-regulate osteogenic and down-regulate adipogenic differentiation of bone marrow stromal cells in ovariectomized rats.

机构信息

The First Clinical Medical College, Zhejiang Traditional Chinese Medical University, China.

出版信息

Orthop Surg. 2011 May;3(2):119-26. doi: 10.1111/j.1757-7861.2011.00129.x.

DOI:10.1111/j.1757-7861.2011.00129.x

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC6583533/

Abstract

OBJECTIVE

To observe the dynamic expression of DKK1 protein in the process whereby Epimedium-derived flavonoids (EFs) regulate the balance between osteogenic and adipogenic differentiation of bone marrow stromal cells in ovariectomized rats, and to provide experimental evidence for the mechanism of EFs in the treatment of postmenopausal osteoporosis.

METHODS

Bone marrow stromal cells from ovariectomized rats were separated and cultivated in osteoinductive or liquid medium for 15 days in vitro. EFs (10 µg/mL) were applied to both cultures. Alkaline phosphatase (ALP) staining, ALP activity determination, Oil Red O staining and fluorescence quantitative polymerase chain reaction were used to determine the influence of EFs on osteogenic and adipogenic differentiation of bone marrow stromal cells in ovariectomized rats. Moreover, in order to explore the exact mechanism of EFs on osteogenic and adipogenic differentiation of bone marrow stromal cells in ovariectomized rats, enzyme linked immunosorbent assay was used to determine the dynamic expression of DKK1 protein in this process.

RESULTS

EFs increased activity of ALP and mRNA expression of Runx2 (early osteoblast differentiation factor) and decreased mRNA expression of PPARγ-2 (key factor of fat generation). Importantly, EFs down-regulated expression of DKK1 protein in an osteogenic induction medium and inhibited up-regulation of DKK1 protein in an adipogenic induction medium.

CONCLUSION

EFs regulate the balance between osteogenic and adipogenic differentiation of bone marrow stromal cells in ovariectomized rats by down-regulating expression of DKK1 protein. This may be an important molecular mechanism of EFs in the context of treatment of postmenopausal osteoporosis.

摘要

目的

观察淫羊藿黄酮（EFs）调节去卵巢大鼠骨髓基质细胞成骨和成脂分化平衡过程中 DKK1 蛋白的动态表达，为 EFs 治疗绝经后骨质疏松症的机制提供实验依据。

方法

体外分离培养去卵巢大鼠骨髓基质细胞，分别在成骨诱导液和液体培养基中培养 15 天。将 EFs（10 μg/ml）分别加入两种培养基中。碱性磷酸酶（ALP）染色、ALP 活性测定、油红 O 染色和荧光定量聚合酶链反应检测 EFs 对去卵巢大鼠骨髓基质细胞成骨和成脂分化的影响。此外，为了探讨 EFs 对去卵巢大鼠骨髓基质细胞成骨和成脂分化的确切机制，采用酶联免疫吸附法检测 DKK1 蛋白在这一过程中的动态表达。

结果

EFs 增加了 ALP 的活性和 Runx2（早期成骨分化因子）mRNA 的表达，降低了 PPARγ-2（脂肪生成的关键因子）mRNA 的表达。重要的是，EFs 在成骨诱导培养基中下调了 DKK1 蛋白的表达，并抑制了在脂肪诱导培养基中 DKK1 蛋白的上调。

结论

EFs 通过下调 DKK1 蛋白的表达来调节去卵巢大鼠骨髓基质细胞成骨和成脂分化的平衡。这可能是 EFs 在治疗绝经后骨质疏松症方面的一个重要分子机制。