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

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.

Suppressive Effects of Rosmarinic Acid Rich Fraction from Perilla on Oxidative Stress, Inflammation and Metastasis Ability in A549 Cells Exposed to PM via C-Jun, P-65-Nf-Κb and Akt Signaling Pathways

“…Our results also demonstrated that RA-RF, a natural polyphenol component found in perilla, reduced IL-6, IL-8, TNF-α, and COX-2 gene expression. This finding is in agreement with our previous report showing that RA extracted from perilla reduced mRNA levels of pro-inflammatory cytokines [20,22,74]. Additionally, RA-rich extract from Trichodesma khasianum leaves could diminish ROS production and inflammatory modulators such as IL-6, TNF-α, and COX-2 levels in vitro and in vivo [75].…”

“…This work resembles previous studies showing that the ethanolic extract of perilla leaves disrupted c-Jun phosphorylation [93]. Moreover, RA-rich extract from perilla seed meal could reduce RANKL-induced c-Jun translocation in RAW macrophage cells [74]. In addition, RA alone exerted protective effects against oxidative stress, inflammation, invasion, migration, and MMP-9 protein expression through the AP-1 signaling cascade [94,95].…”

“…In our study, RA-RF evidently suppressed the phos-phorylation of p65-NF-κB in A549 cells treated with PMFF. Other reports showed that the perilla extract rich in RA [74] and RA compounds had a considerable inhibitory effect on inflammation via suppression of the NF-κB signaling pathway during both in vitro and in vivo experiments [103,104]. Moreover, RA inhibited the invasion and migration of human glioma cells [85] and human hepatoma cells through the PI3K/Akt/NF-κB signaling pathway [105].…”

Molecular Mechanism of Antioxidant and Anti-Inflammatory Effects of Omega-3 Fatty Acids in Perilla Seed Oil and Rosmarinic Acid Rich Fraction Extracted from Perilla Seed Meal on TNF-α Induced A549 Lung Adenocarcinoma Cells

“…These findings are consistent with those reported previously. It was found that cold-pressed PSO mainly consisting of ω-3 fatty acid reduced the ROS production induced by ultraviolet radiation in normal human dermal fibroblasts [53], and the EA fraction from PSM, which contained high quantities of RA, inhibited ROS production in RANKL-induced RAW 264.7 cells [54]. Our results suggest that both PSO and RA-RF have antioxidant ability and equally inhibit ROS generation.…”

Comparison of Phenolic Contents and Scavenging Activities of Miang Extracts Derived from Filamentous and Non-Filamentous Fungi-Based Fermentation Processes

“…The inhibition of intracellular ROS production induced by hydrogen peroxide (H 2 O 2 ) was determined following the method of Saenjum et al [21] and Phromnoi et al [22] with slight modifications. The assay measures the oxidation of 2 ,7 -dichlorodihydrofluorescein diacetate (H 2 DCF-DA) to fluorescent 2 ,7 -dichlorofluorescein (DCF).…”