The effects of Pinoresinol diglucoside on the differentiation and bone resorption of osteoclast RAW264.7

Zhefeng Jin,Hongtao Li,Hongmei Cao,Fangshan Bi

doi:10.1590/fst.89221

Abstract

The paper aimed to investigate the effect of Pinoresinol diglucoside on the differentiation and bone resorption of osteoclast RAW264.7, and to evaluate the effect of Pinoresinol diglucoside on osteoclasts. In order to study the effect of Pinoresinol diglucoside on osteoclast differentiation, RANKL was used to induce RAW264.7 cells to differentiate into osteoclasts, and different concentrations of Pinoresinol diglucoside was added to intervene. CCK-8 method was applied to detect cell viability. TRAP staining was employed to observe cell morphology. Annexin V/PI flow cytometry was used to detect cell apoptosis. Phalloidin was used to detect F-actin formation of osteoclasts and to observe the effect on bone resorption. WB was employed to detect the effects of differentiation-related proteins and RANKL/RANK signaling pathways, and immunofluorescence detection technology was used to measure the distribution and nuclear translocation of p65. Pinoresinol diglucoside can effectively inhibit RANKL-induced osteoclast differentiation, especially in the early stage. The drug can inhibit the formation of F-actin of osteoclasts and inhibit bone resorption. Through inhibiting the ubiquitination and degradation of the homologous phosphatase tensin (PTEN), the drug up-regulated the viability of PTEN. The up-regulated PTEN viability then inhibited the NF-κB and AKT signaling pathways, resulting in a decrease in the expressions of nuclear factor c1 (NFATc1) in activated T cells. Pinoresinol diglucoside effectively inhibited the formation of F-actin and bone resorption in mature osteoclasts. The mechanism is through inhibiting the expression levels of osteoclast-related proteins NFATc1, c-Fos, CSTK and TRAP and RNAKL/RANK signaling pathways, and also inhibiting the activation of NF-κB and AKT signaling pathways.

Highlights

Bone is a tissue that is constantly being renewed and remodeled
They are derived from hematopoietic stem cell lines and are a kind of multinucleated giant cell fused under the stimulation of the macrophage colony stimulating factor (M-CSF) and nuclear factor-κB receptors factor ligand (RANKL) of monocytes/macrophages(Khosla & Hofbauer,2 017; Jia et al, 2018)
NF-κB p65 has a nuclear localization sequence, which can initiate the transcription of downstream related genes after being guided into the nucleus (Choi 2019; Liu et al, 2017) In this study, we found that Pinoresinol diglucoside can inhibit the phosphorylation and degradation of IκBα induced by RANKL, thereby inhibiting the phosphorylation of NF-κB p65

Summary

Introduction

Osteoblasts synthesize bone matrix and osteoclasts absorb bone (Aspray & Hill, 2019). Various small disturbances, such as the levels of inflammatory factors, growth factors and hormones, may cause the delicate balance between osteogenesis and osteoclastogenesis to be broken, leading to bone-related diseases. Excessive activation of osteoclasts is the main cause of the occurrence of these diseases(Anthamatten & Parish, 2019; Tanaka, 2019)Osteoclasts are the only cells that perform bone resorption in the human body. They are derived from hematopoietic stem cell lines and are a kind of multinucleated giant cell fused under the stimulation of the macrophage colony stimulating factor (M-CSF) and nuclear factor-κB receptors factor ligand (RANKL) of monocytes/macrophages(Khosla & Hofbauer,2 017; Jia et al, 2018)

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Discussion

Conclusion