Abstract
One of the Korean endemic plants, Abeliophyllum distichum Nakai (Oleaceae), contains acteoside, which is a glycoside exhibiting neuroprotective, anti-inflammation effects and antibacterial capacities. We conducted an investigation on the effects of the callus of A. distichum (cultivar Okhwang 1, CAO) on pro-inflammatory mediators released following nuclear factor-кB (NF-кB), phosphatidylinositol 3-kinase/Akt (PI3K-Akt) and mitogen-activated protein kinase (MAPK) signal activation in lipopolysaccharide (LPS)-induced RAW 264.7 cells. Immunoblotting was employed to find out the expression of cyclooxygenase-2 (COX-2), inducible nitric oxide (iNOS), and activation of MAPK molecules, NF-κB and Akt. Cytokines, COX-2, and iNOS gene expression were assessed using polymerase chain reaction techniques. Cytokines, COX-2, and iNOS gene expression were assessed using polymerase chain reaction techniques. High-performance liquid chromatography revealed that CAO was rich in acteoside and isoacteoside. As a result, CAO inhibited the generation of NO, cytokines, COX-2, and iNOS expression. Further, translocation to the nuclear of NF-κB p65 and degradation of the inhibitor of NF-кB (IкB) were alleviated by suppressing phosphorylation. Additionally, CAO significantly impacted MAPK pathway activation by potentially reducing phosphorylation of MAPKs. These results indicate that the anti-inflammatory effect of CAO is mediated via the inhibition of MAPK, PI3K/Akt, and NF-κB signaling pathways, probably via glycosides, phenolics, and flavonoids bioactivity derived from plants. CAO can serve as a potential anti-inflammatory agent, which alleviates inflammation factors and act through specific cell signaling pathways.
Highlights
The chromatogram of CAO was identified by comparing the retention time (RT) to that of standard acteoside and isoacteoside at 330 nm
Compared with the standard curve, the amount of acteoside was analyzed to be 399.27 ± 2.3 mg/g (39.93% w/w), and isoacteoside was analyzed to be 56.69 ± 1.1 mg/g (5.67% (w/w) in CAO. These results confirmed that acteoside and isoacteoside, with chemical structures shown in Figure 2C,D, are the major compounds in CAO
2021, 9, These results confirmed that acteoside and isoacteoside, with chemical structures shown in Figure 2C,D, are the major compounds in CAO
Summary
The stimuli are caused by injured cells, pathogens, and endogenous signals, which result in cellular healing to restore normal function [1]. These reactions may repair injured tissues and maintain homeostasis, which is regulated by numerous cytokines and inflammatory mediators, which includes the tumor necrosis factor (TNF)-α, nitric oxide (NO), and interleukin-6 (IL-6) [2,3]. LPS activates macrophages to secrete pro-inflammatory cytokines and reactive oxidative species (ROS) [5,6] Under these stimulations, the pro-inflammatory mediators, including NO and prostaglandin E2 (PGE2), are generated in large amounts by inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2), respectively [7]
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