Abstract

Atopic dermatitis (AD) is a chronic inflammatory skin disease caused by immune hypersensitivity reaction. The cause of AD is unclear, but its symptoms have a negative effect on quality of life; various treatment methods to alleviate these symptoms are underway. In the present study, we aimed to evaluate in vitro antioxidant and anti-inflammatory effects of Rubus coreanus water extract (RCW) on AD. Total phenolic compounds and flavonoid content of RCW were 4242.40 ± 54.84 mg GAE/g RCE and 1010.99 ± 14.75 mg CE/g RCW, respectively. RCW reduced intracellular reactive oxygen species level and increased the action of antioxidant enzymes, such as catalase, superoxide dismutase, and glutathione peroxidase in tumor necrosis factor-α (TNF-α)/interferon-γ (IFN-γ)-stimulated HaCaT cells. Moreover, mRNA expression of the pro-inflammatory cytokines, including TNF-α, interleukin-1β, and interleukin-6, was downregulated by RCW in the TNF-α/IFN-γ-stimulated cells. The levels of inflammatory chemokines (thymus- and activation-regulated chemokine; eotaxin; macrophage-derived chemokine; regulated on activation, normal T-cell expressed and secreted; and granulocyte-macrophage colony-stimulating factor) and intercellular adhesion molecule-1 were decreased in the TNF-α/IFN-γ-stimulated HaCaT cells after RCW treatment. Additionally, the mRNA expression levels of filaggrin and involucrin, proteins that form the skin, were increased by RCW. Furthermore, RCW inhibited the nuclear factor kappa-light-chain-enhancer of the activated B cells pathway in the TNF-α/IFN-γ-stimulated HaCaT cells. Collectively, the present investigation indicates that RCW is a potent substance that inhibits AD.

Highlights

  • Atopic dermatitis (AD) is a chronic inflammatory skin disease with repeated longterm recovery and recurrence [1]

  • By evaluating changes in the expression of inflammation-related cytokines, atopic dermatitisrelated chemokines, and genes related to skin formation proteins, we aimed to confirm the molecular mechanism underlying the anti-inflammatory activity of Rubus coreanus water extract (RCW)

  • The results showed that the total phenolic compounds and flavonoid content in the RCW

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Summary

Introduction

Atopic dermatitis (AD) is a chronic inflammatory skin disease with repeated longterm recovery and recurrence [1]. AD is classified as a type of allergic disease, such as allergic rhinitis and asthma, and presents symptoms such as skin dryness, erythema, edema, pruritus, and desquamation [2] These symptoms are not life-threatening; they reduce the quality of life, but are difficult to treat, as they may occur due to both environmental and genetic factors [3,4]. Granulocyte-macrophage colony-stimulating factor (GM-CSF) is secreted to differentiate precursor cells such as granulocytes and macrophages and acts as a growth factor for leukocytes These chemokines prompt the infiltration of immune cells, such as T lymphocytes, monocytes, and mast cells into the skin lesion, thereby inducing inflammatory skin diseases [9]. By evaluating changes in the expression of inflammation-related cytokines, atopic dermatitisrelated chemokines, and genes related to skin formation proteins, we aimed to confirm the molecular mechanism underlying the anti-inflammatory activity of RCW

Preparation of RCW
Determination of Total Phenolic Compounds and Flavonoid Contents
Cell Culture and Treatment
Assessment of Intracellular ROS Formation
Evaluation of Antioxidant Enzyme Activity
Western Blotting
2.10. Statistical Analysis
Total Phenolic Compounds and Flavonoid Content of RCW
Cytotoxicity of RCW in HaCaT Cells
Conclusions
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