Chrysosplenetin promotes osteoblastogenesis of bone marrow stromal cells via Wnt/β-catenin pathway and enhances osteogenesis in estrogen deficiency-induced bone loss

  • Stem Cell Res Ther. 2019 Aug 29;10(1):277. doi: 10.1186/s13287-019-1375-x.
Guoju Hong  1  2 ,  Xiaoming He  2 ,  Yingshan Shen  2 ,  Xiaojun Chen  2 ,  Fang Yang  2 ,  Peng Yang  2 ,  Fengxiang Pang  2 ,  Xiaorui Han  3 ,  Wei He  4  5 ,  Qiushi Wei  6  7
Affiliations
  • 1. Department of Surgery, The University of Alberta, Edmonton, Alberta, Canada.
  • 2. The National Key Discipline and the Orthopedic Laboratory, Guangzhou University of Chinese Medicine, Guangzhou, Guangdong, People's Republic of China.
  • 3. School of Medicine, South China University of Technology, Guangzhou, Guangdong, People's Republic of China.
  • 4. Department of Orthopedic, The First Affiliated Hospital of Guangzhou University of Chinese Medicine, Guangzhou, Guangdong, People's Republic of China.
  • 5. Hip Preserving Ward, The First Affiliated Hospital of Guangzhou University of Chinese Medicine, No. 3 Orthopaedic Region, Guangzhou, Guangdong, People's Republic of China.
  • 6. Department of Orthopedic, The First Affiliated Hospital of Guangzhou University of Chinese Medicine, Guangzhou, Guangdong, People's Republic of China. [email protected].
  • 7. Hip Preserving Ward, The First Affiliated Hospital of Guangzhou University of Chinese Medicine, No. 3 Orthopaedic Region, Guangzhou, Guangdong, People's Republic of China. [email protected].
PMID: 31464653 DOI: 10.1186/s13287-019-1375-x
Abstract

Background: Chrysosplenetin is an O-methylated flavonol compound isolated from the plant Chamomilla recutita and Laggera pterodonta. The aim of our research is to evaluate the function of Chrysosplenetin on osteogenesis of human-derived bone marrow stromal cells (hBMSCs) and inhibition of estrogen deficiency-induced osteoporosis via the Wnt/β-catenin signaling pathway.

Method: hBMSCs are cultured and treated by Chrysosplenetin in the absence or presence of Wnt Inhibitor dickkopf-related protein 1 (DKK1) or Bone Morphogenetic Protein 2 (BMP2) antagonist Noggin. RT-qPCR is taken to identify the genetic expression of target genes of Wnt/β-catenin pathway and osteoblast-specific markers. The situation of β-catenin is measured by western blot and immunofluorescence staining. An ovariectomized (OVX) mouse model is set up to detect the bone loss suppression by injecting Chrysosplenetin. Micro-CT and histological assay are performed to evaluate the protection of bone matrix and osteoblast number. Serum markers related with osteogenesis are detected by ELISA.

Results: In the present study, it is found that Chrysosplenetin time-dependently promoted proliferation and osteoblastogenesis of hBMSCs reaching its maximal effects at a concentration of 10 μM. The expressions of target genes of Wnt/β-catenin pathway and osteoblast-specific marker genes are enhanced by Chrysosplenetin treatment. Furthermore, the phosphorylation of β-catenin is decreased, and nuclear translocation of β-catenin is promoted by Chrysosplenetin. Osteogenesis effects mentioned above are founded to be blocked by DKK1 or BMP2 antagonist Noggin. In vivo study reveals that Chrysosplenetin prevents estrogen deficiency-induced bone loss in OVX mice detected by Micro-CT, histological analysis, and ELISA.

Conclusions: Our study demonstrates that Chrysosplenetin improves osteoblastogenesis of hBMSCs and osteogenesis in estrogen deficiency-induced bone loss by regulating Wnt/β-catenin pathway.

Keywords
BMSC; Chrysosplenetin; DKK1; Noggin; Osteoblast; Wnt/β-catenin.
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