10-Gingerol Suppresses Osteoclastogenesis in RAW264.7 Cells and Zebrafish Osteoporotic Scales

Abstract

Osteoporosis is the most common aging-associated bone disease and is caused by hyperactivation of osteoclastic activity. We previously reported that the hexane extract of ginger rhizome [ginger hexane extract (GHE)] could suppress receptor activator of nuclear factor kappa-B ligand (RANKL)-induced osteoclastogenesis in RAW264.7 cells. However, the anti-osteoclastic components in GHE have not yet been identified. In this study, we separated GHE into several fractions using silica gel column chromatography and evaluated their effects on osteoclastogenesis using a RAW264.7 cell osteoclast differentiation assay (in vitro) and the zebrafish scale model of osteoporosis (in vivo). We identified that the fractions containing 10-gingerol suppressed osteoclastogenesis in RAW264.7 cells detected by tartrate-resistant acid phosphatase (TRAP) staining. In zebrafish, GHE and 10-gingerol suppressed osteoclastogenesis in prednisolone-induced osteoporosis regenerated scales to promote normal regeneration. Gene expression analysis revealed that 10-gingerol suppressed osteoclast markers in RAW264.7 cells [osteoclast-associated immunoglobulin-like receptor, dendrocyte-expressed seven transmembrane protein, and matrix metallopeptidase-9 (Mmp9)] and zebrafish scales [osteoclast-specific cathepsin K (CTSK), mmp2, and mmp9]. Interestingly, nuclear factor of activated T-cells cytoplasmic 1, a master transcription regulator of osteoclast differentiation upstream of the osteoclastic activators, was downregulated in zebrafish scales but showed no alteration in RAW264.7 cells. In addition, 10-gingerol inhibited CTSK activity under cell-free conditions. This is the first study, to our knowledge, that has found that 10-gingerol in GHE could suppress osteoclastic activity in both in vitro and in vivo conditions.

Potential Anti-osteoporosis compounds from Zingiber officinale: A Molecular Docking and Pharmacokinetics Prediction

“…6-Gingerol was known to have osteogenic activity by increasing osteoblasts' transcription levels, alkaline phosphatase activity, and enhanced mineralized nodule formation in vitro 52 . In addition, 10-gingerol was reported to inhibit osteoclasts activity in vitro, marked with downregulated osteoclastic markers and cathepsin K activity 53 . 10-Gingerol was also found to inhibits osteoclastogenesis in prednisolone-induced osteoporosis in zebrafish animal models 53 .…”

Zebrafish mutants reveal unexpected role of Lrp5 in osteoclast regulation

“…Zebrafish elasmoid scales are translucent mineralized structures of the dermal skeleton with osteoblasts and osteoclasts present (42). Over the last decade zebrafish scales have been used to exhibit the mechanism of bone remodeling (34,43,44). Using Von Kossa staining, we observed a distinctive demineralized area at the base of scales in 77% of lrp5 mutant scales (n=134), compared to just 22% in wildtype siblings (n= 118) (Figure 6A).…”

A zebrafish model of diabetic nephropathy shows hyperglycemia, proteinuria and activation of the PI3K/Akt pathway

“…The transgenic line Tg(l-fabp::VDBP-GFP) ( Zhou and Hildebrandt, 2012 ) and skeletal muscle insulin-resistant zebrafish [zMIR; Tg(acta1:dnIGF1R-EGFP) ] ( Maddison et al, 2015 ) were raised and maintained as previously described ( Zang et al, 2021 , 2022a ) under standard guidelines ( Westerfield, 2007 ). Embryos were collected after the natural spawn, incubated in 0.3× Danieau's solution [17.4 mM NaCl, 0.21 mM KCl, 0.12 mM MgSO 4 , 0.18 mM Ca(NO 3 ) 2 and 1.5 mM HEPES; pH 7.6], and staged as hours post fertilization.…”