Diaporisoindole E inhibits RANKL-induced osteoclastogenesis via suppression of PI3K/AKT and MAPK signal pathways

Phytomedicine. 2020 Aug 15:75:153234. doi: 10.1016/j.phymed.2020.153234. Epub 2020 May 15.

Abstract

Background: Diaporisoindole E (SA8), an isoprenylisoindole alkaloids isolated from the mangrove endophytic fungus Diaporthe sp. SYSU-HQ3, was reported with anti-inflammatory activity in RAW264.7 cells. However, the effect of SA8 in bone metabolism is unknown.

Purpose: The purpose of this study is to investigate the inhibitory effect of SA8 in RANKL-induced osteoclastogenesis and to explore its mechanism of action.

Methods: Osteoclastogenesis was assayed by TRAP staining. Expression of osteoclast specific genes was evaluated by real time-PCR. The inhibition of phosphorylation of the protein was measured by western blot analysis. The transcription activity of NF-κB was conducted using luciferase reporter gene assays. Osteoblast differentiation was assayed by alkaline phosphatase and Alizarin Red staining.

Results: SA8 significantly inhibited the osteoclast differentiation in a dose- and time-dependent manner, which is consistent with the suppression of osteoclast specific genes including TRAP, DC-stamp, NFATc1, MMP-9, and ATP6v0d2. Further study on the mechanism of action revealed that SA8 inhibited osteoclast differentiation by attenuating PI3K/AKT and MAPK but not through NF-κB signaling pathways. Moreover, SA8 also suppressed bone resorption activity in a hydroxyapatite-coated plate without affecting osteoblast differentiation in C3H10T1/2 using alkaline phosphatase and Alizarin Red staining.

Conclusions: These findings suggest that SA8 (Diaporisoindole E) is the potential anti-osteoporosis agent.

Keywords: Diaporisoindole E; MAPK; Osteoclastogenesis; Osteoporosis; PI3K/AKT.

MeSH terms

  • Animals
  • Bone and Bones / cytology
  • Bone and Bones / metabolism
  • Cell Differentiation / drug effects
  • Gene Expression Regulation / drug effects
  • Indole Alkaloids / pharmacology*
  • MAP Kinase Signaling System / drug effects
  • Mice
  • Mice, Inbred C57BL
  • NF-kappa B / genetics
  • NF-kappa B / metabolism
  • Osteoclasts / drug effects*
  • Osteoclasts / physiology
  • Osteogenesis / drug effects*
  • Phosphatidylinositol 3-Kinases / metabolism*
  • Phosphorylation / drug effects
  • Proto-Oncogene Proteins c-akt / metabolism*
  • RANK Ligand / metabolism*
  • RAW 264.7 Cells
  • Signal Transduction / drug effects

Substances

  • Indole Alkaloids
  • NF-kappa B
  • RANK Ligand
  • Tnfsf11 protein, mouse
  • Proto-Oncogene Proteins c-akt