First Clinical Medical College, Guangzhou University of Chinese Medicine, Guangzhou, PR China.
Hip Preserving Ward, No. 3 Orthopaedic Region, The First Affiliated Hospital of Guangzhou University of Chinese Medicine, Guangzhou, PR China; Institute of Hip Joint, Guangzhou University of Chinese Medicine, Guangzhou, PR China.
Biomed Pharmacother. 2019 Apr;112:108746. doi: 10.1016/j.biopha.2019.108746. Epub 2019 Mar 2.
Steroid-induced osteonecrosis of the femoral head (SONFH) is a refractory disease induced by glucocorticoids. Marrow mesenchymal stem cells (MSCs) differentiate into multiple bone matrix cells and have been used as cell-based therapies to treat ONFH. However, the osteogenesis of MSCs isolated from patients with SONFH is significantly decreased. Polydatin has been widely used in traditional Chinese remedies due to its multiple pharmacological actions. As shown in our previous study, Polydatin protects from oxidative stress and promotes BMSC migration. However, little is known about its role in BMSC (Bone marrow mesenchymal stem cells) osteogenesis; therefore, we further investigated the effect and mechanism of Polydatin in hBMSC osteogenesis. The ability of Polydatin to promote the proliferation and osteogenic differentiation of hBMSCs was determined using the MTT assay, ALP staining and the ALP activity assay. Next, qPCR and western blotting were performed to measure the levels of genes and proteins related to the osteogenesis of hBMSCs. Then, the effect of Polydatin on the nuclear translocation of β-catenin was determined using immunofluorescence staining. Polydatin (30 μM) markedly enhanced the proliferation of hBMSCs and alkaline phosphatase (ALP) activity. Additionally, it also significantly upregulated the expression of osteogenic genes (Runx2, osteopontin, DLX5, osteocalcin, collagen type I and BMP2) and components of the Wnt signaling pathway (β-catenin, Lef1, TCF7, c-jun, c-myc and cyclin D). These osteogenesis-potentiating effects of Polydatin were blocked by Noggin, an inhibitor of the BMP pathway, and DKK1, an inhibitor of the Wnt/β-catenin pathway. However, DKK1 did not affect Polydatin-induced BMP2 expression. Based on our results, Polydatin promotes the proliferation and osteogenic differentiation of hBMSCs through the BMP2-Wnt/β-catenin signaling pathway.
激素诱导性股骨头坏死（SONFH）是一种由糖皮质激素诱导的难治性疾病。骨髓间充质干细胞（MSCs）可分化为多种骨基质细胞，已被用作细胞疗法来治疗 ONFH。然而，从 SONFH 患者中分离的 MSCs 的成骨能力显著降低。虎杖苷由于其多种药理作用而被广泛用于中药方剂。如我们之前的研究所示，虎杖苷可抵抗氧化应激并促进 BMSC 迁移。然而，其在 BMSC（骨髓间充质干细胞）成骨中的作用知之甚少；因此，我们进一步研究了虎杖苷对 hBMSC 成骨的作用及其机制。通过 MTT 检测、碱性磷酸酶（ALP）染色和 ALP 活性检测来确定虎杖苷对 hBMSC 增殖和成骨分化的能力。接下来，通过 qPCR 和 Western blot 检测来测量与 hBMSC 成骨相关的基因和蛋白的水平。然后，通过免疫荧光染色来检测虎杖苷对 β-catenin 核易位的影响。虎杖苷（30 μM）显著增强了 hBMSC 的增殖和碱性磷酸酶（ALP）活性。此外，它还显著上调了成骨基因（Runx2、骨桥蛋白、DLX5、骨钙素、I 型胶原和 BMP2）和 Wnt 信号通路成分（β-catenin、Lef1、TCF7、c-jun、c-myc 和 cyclin D）的表达。BMP 通路抑制剂 Noggin 和 Wnt/β-catenin 通路抑制剂 DKK1 阻断了虎杖苷的这些促骨生成作用。然而，DKK1 不影响虎杖苷诱导的 BMP2 表达。基于我们的结果，虎杖苷通过 BMP2-Wnt/β-catenin 信号通路促进 hBMSC 的增殖和成骨分化。
