Fisetin, a 3,7,3′,4′-Tetrahydroxyflavone Inhibits the PI3K/Akt/mTOR and MAPK Pathways and Ameliorates Psoriasis Pathology in 2D and 3D Organotypic Human Inflammatory Skin Models

Abstract

Psoriasis is a chronic immune-mediated skin disease that involves the interaction of immune and skin cells, and is characterized by cytokine-driven epidermal hyperplasia, deviant differentiation, inflammation, and angiogenesis. Because the available treatments for psoriasis have significant limitations, dietary products are potential natural sources of therapeutic molecules, which can repair the molecular defects associated with psoriasis and could possibly be developed for its management. Fisetin (3,7,3′,4′-tetrahydroxyflavone), a phytochemical naturally found in pigmented fruits and vegetables, has demonstrated proapoptotic and antioxidant effects in several malignancies. This study utilized biochemical, cellular, pharmacological, and tissue engineering tools to characterize the effects of fisetin on normal human epidermal keratinocytes (NHEKs), peripheral blood mononuclear cells (PBMC), and CD4+ T lymphocytes in 2D and 3D psoriasis-like disease models. Fisetin treatment of NHEKs dose- and time-dependently induced differentiation and inhibited interleukin-22-induced proliferation, as well as activation of the PI3K/Akt/mTOR pathway. Fisetin treatment of TNF-α stimulated NHEKs also significantly inhibited the activation of p38 and JNK, but had enhanced effect on ERK1/2 (MAPK). In addition, fisetin treatment significantly decreased the secretion of Th1/Th-17 pro-inflammatory cytokines, particularly IFN-γ and IL-17A by 12-O-tetradecanolylphorbol 13-acetate (TPA)-stimulated NHEKs and anti-CD3/CD28-activated human PBMCs. Furthermore, we established the in vivo relevance of fisetin functions, using a 3D full-thickness human skin model of psoriasis (FTRHSP) that closely mimics in vivo human psoriatic skin lesions. Herein, fisetin significantly ameliorated psoriasis-like disease features, and decreased the production of IL-17 by CD4+ T lymphocytes co-cultured with FTRHSP. Collectively, our data identify the prodifferentiative, antiproliferative, and anti-inflammatory effects of fisetin, via modulation of the PI3K-Akt-mTOR and p38/JNK pathways and the production of cytokines in 2D and 3D human skin models of psoriasis. These results suggest that fisetin has a great potential to be developed as an effective and inexpensive agent for the treatment of psoriasis and other related inflammatory skin disorders.

Natural Compound Mixture, Containing Emodin, Genipin, Chlorogenic Acid, Cimigenoside, and Ginsenoside Rb1, Ameliorates Psoriasis‐Like Skin Lesions by Suppressing Inflammation and Proliferation in Keratinocytes

“…IL-22 expression is upregulated in the skin lesions and serum of patients with psoriasis, and it is strongly related to the disease severity [ 47 ]. IL-22-induced keratinocyte hyperproliferation is regulated by the PI3K/Akt/mTOR signaling pathway, and targeting this pathway is a potent approach for psoriasis treatment [ 21 , 48 ]. Histologically, PSM treatment ameliorated epidermal hyperplasia significantly in IMQ-induced C57BL/6 mice.…”

Identification of new fisetin analogs as kinase inhibitors: Data on synthesis and anti-skin cancer activities evaluation

“…Cell cultures were prepared, analyzed, and protein content was quantified using the BCA reagent assay (Thermo Fisher Scientific, Waltham, MA, USA). Protein electrophoresis: SDS-PAGE method (Protean 3 system, Bio-Rad, Hercules, CA, USA), followed by protein expression via the western blotting method, using the Bio-Rad analysis and imaging system as earlier described [1 , 2] . Analysis and graphs in the figures, including in vitro enzymatic kinase activities, western blot, scratch wound assays, and colony formation were plotted using GraphPad PRISM program suite, version 8 (GraphPad Software, Inc., La Jolla, CA, USA), and molecular docking analysis [ 1 , 2] .…”

“…Protein electrophoresis: SDS-PAGE method (Protean 3 system, Bio-Rad, Hercules, CA, USA), followed by protein expression via the western blotting method, using the Bio-Rad analysis and imaging system as earlier described [1 , 2] . Analysis and graphs in the figures, including in vitro enzymatic kinase activities, western blot, scratch wound assays, and colony formation were plotted using GraphPad PRISM program suite, version 8 (GraphPad Software, Inc., La Jolla, CA, USA), and molecular docking analysis [ 1 , 2] . In silico prediction of skin permeation, lipophilicity, absorption, distribution, metabolism, excretion (ADMET) properties of the analogs compared to the reference drug fisetin were performed and analyzed using the online SwissADME platform of the Swiss Institute of Bioinformatics [3] .…”

Tissue engineering to better understand senescence: Organotypics come of age

“…Tissue engineering allows the culture of multiple cell types in a tightly controlled environment, and has previously been used to study immune cells and their mechanisms of inflammation. Activated T-cells (CD4+ cells) in an LSE constructed with normal human keratinocytes can cause a psoriatic phenotype, including hyperproliferation, reflective of the inflammatory state of psoriatic skin ( Chamcheu et al, 2019 ). This work also validated the ability of fisetin, a compound of interest to the senescence field due to its capacity to selectively kill senescent cells ( Yousefzadeh et al, 2018 ), to treat psoriatic symptoms by suppressing the inflammatory response ( Chamcheu et al, 2019 ).…”

“…Activated T-cells (CD4+ cells) in an LSE constructed with normal human keratinocytes can cause a psoriatic phenotype, including hyperproliferation, reflective of the inflammatory state of psoriatic skin ( Chamcheu et al, 2019 ). This work also validated the ability of fisetin, a compound of interest to the senescence field due to its capacity to selectively kill senescent cells ( Yousefzadeh et al, 2018 ), to treat psoriatic symptoms by suppressing the inflammatory response ( Chamcheu et al, 2019 ). Local inflammation in LSEs has been demonstrated by exposure to histamine, which caused a loss of keratinocyte differentiation markers and tight junction proteins, suggesting a mechanism by which histamine may contribute to inflammatory skin disorders by causing a defective skin barrier ( Gschwandtner et al, 2013 ).…”