山核桃提取物通过 BMP2/Smad/Runx2 和 Wnt/β-catenin 通路促进人骨髓间充质干细胞的成骨作用。

Juglans regia L. extract promotes osteogenesis of human bone marrow mesenchymal stem cells through BMP2/Smad/Runx2 and Wnt/β-catenin pathways.

机构信息

Health Management Center, The Affiliated Hospital (TCM) of Southwest Medicial University, No. 182, Chunhui Road, Longmatan District, Luzhou, 646000, Sichuan, China.

Sichuan Provincial People's Hospital, Chengdu, 610000, Sichuan, China.

出版信息

J Orthop Surg Res. 2022 Feb 14;17(1):88. doi: 10.1186/s13018-022-02949-1.

BACKGROUND

The present study investigates the effects of Juglans regia L. (walnut, JRL) leaves extract on osteogenesis of human bone marrow mesenchymal stem cells (hBMSCs).

METHODS

hBMSCs were incubated with different concentrations of JRL extract (10, 20, 40, or 80 μM). Cell proliferation was evaluated by Cell Counting Kit-8 assay (CCK-8) assay. ALP activity and Alizarin Red staining were used to assess the osteogenesis of BMSCs. Western blot was performed to measure the levels of proteins.

RESULTS

Our results showed all concentrations of JRL extract had no significant effect on cell proliferation. JRL extract concentration-dependently promoted osteoblastic differentiation and cell autophagy of hBMSCs, characterized by the increased expression of pro-osteogenic markers alkaline phosphatase (ALP), osteocalcin (BGLAP), osterin, and osteoprotegerin (OPG) and autophagy marker proteins (LC3II, Beclin-1, and p62). Furthermore, JRL extract stimulated the activation BMP2/Smad/Runx2 and Wnt/β-catenin signaling pathways in hBMSCs, which play key roles in osteogenesis differentiation. Meanwhile, BMP inhibitor (Noggin) and Wnt antagonist Dickkopf-1 (DKK1) both reversed the increases of BGLAP, osterin, and OPG expression induced by JRL extract.

CONCLUSIONS

Our findings indicate that JRL extract regulated osteogenic differentiation and cell autophagy of hBMSCs through the BMP2/Smad/Runx2 and Wnt/β-catenin pathways.

背景

本研究探讨了胡桃（核桃，JRL）叶提取物对人骨髓间充质干细胞（hBMSCs）成骨的影响。

方法

用不同浓度的 JRL 提取物（10、20、40 或 80 μM）孵育 hBMSCs。通过细胞计数试剂盒-8（CCK-8）检测细胞增殖。用碱性磷酸酶（ALP）活性和茜素红染色来评估 BMSCs 的成骨作用。通过 Western blot 来测量蛋白水平。

结果

我们的结果表明，JRL 提取物的所有浓度对细胞增殖均无显著影响。JRL 提取物浓度依赖性地促进 hBMSCs 的成骨分化和细胞自噬，表现为前成骨标志物碱性磷酸酶（ALP）、骨钙素（BGLAP）、osterin 和骨保护素（OPG）以及自噬标志物蛋白（LC3II、Beclin-1 和 p62）的表达增加。此外，JRL 提取物刺激了 hBMSCs 中 BMP2/Smad/Runx2 和 Wnt/β-catenin 信号通路的激活，这些信号通路在成骨分化中发挥关键作用。同时，BMP 抑制剂（Noggin）和 Wnt 拮抗剂 Dickkopf-1（DKK1）均逆转了 JRL 提取物诱导的 BGLAP、osterin 和 OPG 表达增加。