Biotechnology and Bioprocess Engineering 2021; 26(1): 93-106  
Hydrophobic Fractions of Triticum aestivum L. Extracts Contain Polyphenols and Alleviate Inflammation by Regulating Nuclear Factor-kappa B
Moon-Hee Choi, Min Young Lee, Seung-Hwa Yang, Hyun-Jae Shin, and Young Jin Jeon
Moon-Hee Choi, Seung-Hwa Yang, Hyun-Jae Shin†*
Department of Biochemical and Polymer Engineering, Chosun University, Gwangju 61452, Korea
Tel: +82-62-230-7518; Fax: +82-62-230-7226
E-mail: shinhj@chosun.ac.kr
Min Young Lee, Young Jin Jeon†*
Department of Pharmacology, School of Medicine, Chosun University, Gwangju 61452, Korea
E-mail: yjjeon@chosun.ac.kr
†Equal contribution.
Received: November 2, 2020; Revised: November 11, 2020; Accepted: November 13, 2020; Published online: February 28, 2021.
© The Korean Society for Biotechnology and Bioengineering. All rights reserved.

This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/3.0/) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.
Abstract
As a nutritional supplement, wheatgrass (Triticum aestivum L.) is well-known for its anti-inflammatory effects that are exerted through signal transduction pathways. Wheatgrass was extracted in 80% ethanol (WGE), and the ethanol extracts were further fractionated using ethyl acetate, n-butanol, and water. To determine the antioxidant activity, WGE was tested using various radical-scavenging assays. To explain the anti-inflammatory mechanisms, western blotting, reverse-transcription polymerase chain reaction, luciferase assay, immunofluorescence staining, and the inhibition of mitogen-activated protein kinase (MAPK) phosphorylation were performed using mouse macrophage cells (RAW 264.7). Inhibition of proinflammatory and Th2 cytokines was analyzed using a 2,4-dinitrochlorobenzeneinduced contact dermatitis mouse model. WGE exhibited a strong antioxidant activity in radical scavenging-assays conducted using artificial and natural substrates. The ethyl acetate and n-butanol fractions (hydrophobic fractions) obtained from the 80% ethanolic extract, inhibited the expression of inducible nitric oxide synthase gene, whereas the aqueous layer (the hydrophilic fraction) showed no inhibitory effects. In addition, the hydrophobic fractions inhibited nuclear factor-kappa B (NF-κB) and reduced phosphorylation of MAPK, indicating that inflammation was alleviated possibly by the regulation of NF-κB signaling. This mechanism was further elucidated using the RAW 264.7 cell line and mouse model. Using high-performance liquid chromatography, the hydrophobic fractions were separated into over 10 phenolic compounds, among which benzoic acid, quercetin, and luteolin were abundant. The study demonstrated that wheatgrass is a potential nutraceutical candidate with antioxidant and anti-inflammatory properties for future use in health foods and cosmetics.
Keywords: wheatgrass, Triticum aestivum L., antioxidative polyphenols, hydrophobic fractions, anti-inflammatory nutraceutical

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