Eupafolin ameliorates COX-2 expression and PGE2 production in particulate pollutants-exposed human keratinocytes through ROS/MAPKs pathways

Chiang-Wen Lee; Zih-Chan Lin; Lee-Fen Hsu; Jia-You Fang; Yao-Chang Chiang; Ming-Horng Tsai; Ming-Hsueh Lee; Shu-Yu Li; Stephen Chu-Sung Hu; I-Ta Lee; Feng-Lin Yen

doi:10.1016/j.jep.2016.05.002

Eupafolin ameliorates COX-2 expression and PGE2 production in particulate pollutants-exposed human keratinocytes through ROS/MAPKs pathways

J Ethnopharmacol. 2016 Aug 2:189:300-9. doi: 10.1016/j.jep.2016.05.002. Epub 2016 May 13.

Authors

Chiang-Wen Lee¹, Zih-Chan Lin², Lee-Fen Hsu³, Jia-You Fang⁴, Yao-Chang Chiang⁵, Ming-Horng Tsai⁶, Ming-Hsueh Lee⁷, Shu-Yu Li⁸, Stephen Chu-Sung Hu⁹, I-Ta Lee¹⁰, Feng-Lin Yen¹¹

Affiliations

¹ Division of Basic Medical Sciences, Department of Nursing, Chang Gung Institute of Technology and Chronic Diseases and Health Promotion Research Center, Chiayi, Taiwan; Research Center for Industry of Human Ecology, Chang Gung University of Science and Technology, Kweishan, Taoyuan Taiwan.
² Graduate Institute of BioMedical Sciences, Chang Gung University, Kweishan, Taoyuan, Taiwan; Pharmaceutics Laboratory, Graduate Institute of Natural Products, Chang Gung University, Kweishan, Taoyuan, Taiwan.
³ Department of Respiratory Care, Chang Gung University of Science and Technology, Chiayi Campus, Chiayi, Taiwan.
⁴ Research Center for Industry of Human Ecology, Chang Gung University of Science and Technology, Kweishan, Taoyuan Taiwan; Pharmaceutics Laboratory, Graduate Institute of Natural Products, Chang Gung University, Kweishan, Taoyuan, Taiwan; Department of Anesthetics, Chang Gung Memorial Hospital at Lin-Kou and College of Medicine, Chang Gung University, Kwei-San, Tao-Yuan, Taiwan.
⁵ Center for Drug Abuse and Addiction, China Medical University Hospital, China Medical University, Taichung, Taiwan.
⁶ Department of Pediatrics, Division of Neonatology and Pediatric Hematology/Oncology, Chang Gung Memorial Hospital, Yunlin, Taiwan.
⁷ Division of Neurosurgery, Department of Surgery, Chang Gung Memorial Hospital, Chia-Yi 613 Taiwan.
⁸ Department of Pharmacy, College of Pharmacy & Health Care, Tajen University, Taiwan.
⁹ Department of Dermatology, College of Medicine, Kaohsiung Medical University, Kaohsiung, Taiwan; Department of Dermatology, Kaohsiung Medical University Hospital, Kaohsiung, Taiwan. Electronic address: stephenhu30@hotmail.com.
¹⁰ School of Medicine, College of Medicine, China Medical University, Taichung, Taiwan. Electronic address: ita0128@mail.cmu.edu.tw.
¹¹ Department of Fragrance and Cosmetic Science, College of Pharmacy, Kaohsiung Medical University, No. 100, Tzyou 1st Road, Kaohsiung 807, Taiwan; Institute of Biomedical Sciences, National Sun Yat-Sen University, Kaohsiung, Taiwan. Electronic address: flyen@kmu.edu.tw.

PMID: 27180879
DOI: 10.1016/j.jep.2016.05.002

Abstract

Ethnopharmacological relevance: Eupafolin is a major bioactive compound derived from the methanolic extract of the medicinal herb Phyla nodiflora, which has been used in traditional Chinese medicine to treat various inflammatory diseases. Recently, particulate air pollutants have been shown to induce inflammation of the skin. In this study, we seek to determine whether eupafolin can inhibit the production of inflammatory mediators in a human skin keratinocyte cell line exposed to particulate air pollutants (particulate matter, PM), and determine the molecular mechanisms involved.

Materials and methods: Human keratinocyte HaCaT cells were treated with PM in the presence or absence of eupafolin. Cyclooxygenase-2 (COX-2) protein and gene expression levels were determined by Western blotting, RT-PCR and luciferase activity assay. Prostaglandin E2 (PGE2) production was evaluated by the enzyme immunoassay method. Generation of intracellular reactive oxygen species (ROS) was measured by the dichlorofluorescin (DCFH) oxidation assay, and nicotinamide adenine dinucleotide phosphate (NADPH) oxidase activity was determined by a chemiluminescence assay. For in vivo studies, COX-2 expression in the skin of BALB/c nude mice was analyzed by immunohistochemistry.

Results: Eupafolin inhibited PM-induced COX-2 protein and gene expression and PGE2 production in HaCaT cells. In addition, eupafolin suppressed PM-induced intracellular ROS generation, NADPH oxidase activity, MAPK (ERK, JNK and p38) activation and NK-κB activation. In vivo studies showed that topical treatment with eupafolin inhibited COX-2 expression in the epidermal keratinocytes of PM-treated mice.

Conclusions: Eupafolin exerts anti-inflammatory and antioxidant effects on skin keratinocytes exposed to particulate air pollutants, and may have potential use in the treatment or prevention of air pollutant-induced inflammatory skin diseases in the future.

Keywords: Eupafolin; Eupafolin (PubChem CID: 5317284); Phyla nodiflora; cyclooxygenase-2; keratinocytes; particulate matter; prostaglandin E(2).

MeSH terms

Animals
Anti-Inflammatory Agents / pharmacology*
Cell Line
Cyclooxygenase 2 / genetics
Cyclooxygenase 2 / metabolism*
Cytoprotection / drug effects
Dinoprostone / metabolism*
Down-Regulation
Epidermis / drug effects
Epidermis / enzymology
Flavones / pharmacology*
Humans
Keratinocytes / drug effects*
Keratinocytes / enzymology
Keratinocytes / pathology
Male
Mice, Inbred BALB C
Mice, Nude
Mitogen-Activated Protein Kinases / metabolism*
NF-kappa B / metabolism
Particulate Matter / toxicity*
Phosphorylation
Reactive Oxygen Species / metabolism*
Signal Transduction / drug effects

Substances

Anti-Inflammatory Agents
Flavones
NF-kappa B
Particulate Matter
Reactive Oxygen Species
Ptgs2 protein, mouse
Cyclooxygenase 2
PTGS2 protein, human
Mitogen-Activated Protein Kinases
eupafolin
Dinoprostone