Inhibitory Effect of a Rosmarinic Acid-Enriched Fraction Prepared from Nga-Mon (Perilla frutescens) Seed Meal on Osteoclastogenesis through the RANK Signaling Pathway.

Kanokkarn Phromnoi,Chalermpong Saenjum,Pornngarm Limtrakul (Dejkriengkraikul),Maitree Suttajit

doi:10.3390/antiox10020307

Kanokkarn Phromnoi, Chalermpong Saenjum + Show 2 more

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https://doi.org/10.3390/antiox10020307

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Journal: Antioxidants	Publication Date: Feb 17, 2021
Citations: 24	License type: CC BY 4.0

Affiliation: University of Phayao, Chiang Mai University

Abstract

The aim of this study is to determine antioxidant and anti-inflammatory activities relating to the antiosteoporosis effects of various perilla seed meal (PSM) fractions. The remaining waste of perilla seed obtained from cold oil compression was extracted with 70% ethanol and sequentially fractionated according to solvent polarity with hexane, dichloromethane, ethyl acetate, and water. The results indicated that the seed-meal ethyl acetate fraction (SMEF) exhibited the highest antioxidant and anti-inflammatory activities, and rosmarinic acid (RA) content. The signaling pathways induced by the receptor activator of the nuclear factor kappa B (NF-κB) ligand (RANKL) that trigger reactive oxygen species (ROS) and several transcription factors, leading to the induction of osteoclastogenesis, were also investigated. The SMEF clearly showed attenuated RANKL-induced tartrate-resistant acid phosphatase (TRAP)-positive multinucleated osteoclasts and TRAP activity. A Western blot analysis showed that the SMEF significantly downregulated RANKL-induced NF-κB, AP-1 activation, and the nuclear factor of activated T-cell 1 (NFATc1) expression. SMEF also suppressed RANKL-induced osteoclast-specific marker gene-like MMP-9 using zymography. Furthermore, the SMEF showed inhibition of RANKL-induced ROS production in RAW 264.7 cells. The results suggest that the SMEF, which contained high quantities of RA, could be developed as a natural active pharmaceutical ingredient for osteoclastogenic protection and health promotion.

Highlights

Bone homeostasis is associated with the balance between osteoclast-mediated bone resorption and osteoblast-induced bone formation
We studied the dose-dependent manner of seed-meal ethyl acetate fraction (SMEF) on receptor activator of nuclear factor kappa-B ligand (RANKL)-induced NF-κB, AP-1, and nuclear factor of activated T-cell 1 (NFATc1) expression; cells were pretreated with various concentrations of SMEF
Oxidative stress and antioxidants are associated with the bone remodeling process [3,36]; the antioxidant capacity of perilla seed meal (PSM) fractions was investigated through free-radical-scavenging activity on DPPH, ABTS, O2, and Nitric Oxide (NO) radicals

Summary

Introduction

Bone homeostasis is associated with the balance between osteoclast-mediated bone resorption and osteoblast-induced bone formation. Osteoporosis is a highly prevalent disease that has become a major public health problem [2] and is considered to be associated with chronic low-grade systemic inflammation and oxidative damage [3,4,5]. Bone inflammation induces the production of pro-osteoclastogenic cytokines; namely, the receptor activator of nuclear factor kappa-B ligand (RANKL), tumor necrosis factor-α (TNF-α), and interleukins (IL). It can induce the production of oxidative stress such as reactive oxygen species (ROS) and reactive nitrogen species (RNS), which can alter the bone-remodeling. Antioxidants 2021, 10, 307 process, and make bones weak and fractured [6]. Osteoclasts are multinucleated cells that are differentiated from monocyte–macrophage lineage precursor cells, and are mainly triggered by two hematopoietic factors; namely, macrophage colony-stimulating factor (M-CSF) and RANKL [7,8]

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