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

  • J Orthop Surg Res. 2022 Feb 14;17(1):88. doi: 10.1186/s13018-022-02949-1.
Xianlun Pang  1 ,  Zhendong Zhong  2 ,  Feng Jiang  3 ,  Jian Yang  4 ,  Hai Nie  5
Affiliations
  • 1. Health Management Center, The Affiliated Hospital (TCM) of Southwest Medicial University, No. 182, Chunhui Road, Longmatan District, Luzhou, 646000, Sichuan, China. [email protected].
  • 2. Sichuan Provincial People's Hospital, Chengdu, 610000, Sichuan, China.
  • 3. Department of Orthopedics, The Affiliated Hospital (TCM) of Southwest Medicial University, Luzhou, 646000, Sichuan, China.
  • 4. Department of Orthopedics, Yongchuan Hospital Affiliated to Chongqing Medicial University, Chongqing, 402160, China.
  • 5. Department of Orthopedic Surgery, Sichuan Academy of Medical Sciences and Sichuan Provincial People's Hospital East Campus, Chengdu, 610101, Sichuan, China.
PMID: 35164786 DOI: 10.1186/s13018-022-02949-1
Abstract

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.

Keywords
Autophagy; BMP2/Smad/Runx2; Juglans regia L.; Osteogenic differentiation; Wnt/β-catenin; hBMSCs.
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