Polymethoxyflavones from Kaempferia parviflora ameliorate skin aging in primary human dermal fibroblasts and ex vivo human skin

Abstract Background: Both TGFb and estradiol (E2), a form of estrogen, are pro-fibrotic in the skin. In the connective tissue disease, systemic sclerosis (SSc), both TGFb and E2 are likely pathogenic. Yet, the regulation of TGFb in E2-induced dermal fibrosis remains ill-defined. Elucidating those regulatory mechanisms will improve the understanding of fibrotic disease pathogenesis and set the stage for developing potential therapeutics. Using E2-stimulated primary human dermal fibroblasts in vitro and human skin tissue ex vivo, we identified the important regulatory proteins for TGFb and investigated the extracellular matrix (ECM) components that are directly stimulated by E2-induced TGFb signaling.Methods: We used primary human dermal fibroblasts in vitro and human skin tissue ex vivo stimulated with E2 or vehicle (ethanol) to measure TGFb1, TGFb2 levels using quantitative PCR (qPCR). To identify the necessary cell signaling proteins in E2-induced TGFb1 and TGFb2 transcription, human dermal fibroblasts were pre-treated with an inhibitor of the extracellular signal-regulated kinase/ mitogen-activated protein kinase (ERK/MAPK) pathway, U0126. Finally, human skin tissue ex vivo was pre-treated with SB-431542, a TGFb receptor inhibitor, and ICI 182,780, an estrogen receptor α (ER α) inhibitor, to establish the effects of TGFb and ER α signaling on E2-induced collagen 22A1 (Col22A1) transcription. Results: We found that expression of TGFb1, TGFb2, and Col22A1, a TGFb-responsive gene, are induced in response to E2 stimulation. Mechanistically, Col22A1 induction was blocked by SB-431542 and ICI 182,780 despite E2 stimulation. Additionally, inhibiting E2-induced ERK/MAPK activation and early growth response 1 (EGR1) transcription prevents the E2-induced increase in TGFb1 and TGFb2 transcription and translation. Conclusions: We conclude that E2-induced dermal fibrosis occurs in part through induction of TGFb1, 2 and Col22A1, which is regulated through EGR1 and the MAPK pathway. Thus, blocking estrogen signaling and/or production may be a novel therapeutic option in pro-fibrotic diseases.

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Elucidating the Cellular Mechanism for E2-induced Dermal Fibrosis

10.21203/rs.3.rs-103581/v1 ◽

2020 ◽

Author(s):

DeAnna Baker Frost ◽

Alisa Savchenko ◽

Adeyemi Ogunleye ◽

Milton Armstrong ◽

Carol Feghali-Bostwick

Keyword(s):

Human Skin ◽

Ex Vivo ◽

Mapk Pathway ◽

Dermal Fibroblasts ◽

Skin Tissue ◽

Human Dermal Fibroblasts ◽

Dermal Fibrosis ◽

Tgfb Signaling ◽

Human Skin Tissue

Abstract Background: Both TGFb and estradiol (E2), a form of estrogen, are pro-fibrotic in the skin. In the connective tissue disease, systemic sclerosis (SSc), both TGFb and E2 are likely pathogenic. Yet, the regulation of TGFb in E2-induced dermal fibrosis remains ill-defined. Elucidating those regulatory mechanisms will improve the understanding of fibrotic disease pathogenesis and set the stage for developing potential therapeutics. Using E2-stimulated primary human dermal fibroblasts in vitro and human skin tissue ex vivo, we identified the important regulatory proteins for TGFb and investigated the extracellular matrix (ECM) components that are directly stimulated by E2-induced TGFb signaling.Methods: We used primary human dermal fibroblasts in vitro and human skin tissue ex vivo stimulated with E2 or vehicle (ethanol) to measure TGFb1, TGFb2 levels using quantitative PCR (qPCR). To identify the necessary cell signaling proteins in E2-induced TGFb1 and TGFb2 transcription, human dermal fibroblasts were pre-treated with an inhibitor of the extracellular signal-regulated kinase/ mitogen-activated protein kinase (ERK/MAPK) pathway, U0126. Finally, human skin tissue ex vivo was pre-treated with SB-431542, a TGFb receptor inhibitor, to establish the effects of TGFb signaling on E2-induced collagen 22A1 (Col22A1) transcription. Statistical tests used included parametric and non-parametric ANOVA, student’s T-test and Wilcoxon matched-pairs signed rank test. Significance was defined as p-value ≤ 0.05.Results: We found that TGFb1, TGFb2, and collagen 22A1 (Col22A1), a TGFb-responsive gene, are induced in response to E2 stimulation. Mechanistically, Col22A1 induction is blocked by the TGFb receptor inhibitor SB-431542, despite E2 stimulation. Additionally, inhibiting E2-induced ERK/MAPK activation and early growth response 1 (EGR1) transcription prevents E2-induced increase in TGFb1 and TGFb2 transcription and translation. Conclusions: We conclude that E2-induced dermal fibrosis occurs in part through induction of TGFb1, 2 and Col22A1, which is regulated through EGR1 and the MAPK pathway. Thus, blocking estrogen signaling and/or production may be a novel therapeutic option in pro-fibrotic diseases.

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Elucidating the cellular mechanism for E2-induced dermal fibrosis

Arthritis Research & Therapy ◽

10.1186/s13075-021-02441-x ◽

2021 ◽

Vol 23 (1) ◽

Author(s):

DeAnna Baker Frost ◽

Alisa Savchenko ◽

Adeyemi Ogunleye ◽

Milton Armstrong ◽

Carol Feghali-Bostwick

Keyword(s):

Human Skin ◽

Ex Vivo ◽

Mapk Pathway ◽

Dermal Fibroblasts ◽

Skin Tissue ◽

Human Dermal Fibroblasts ◽

Ici 182,780 ◽

Dermal Fibrosis ◽

Human Skin Tissue

Abstract Background Both TGFβ and estradiol (E2), a form of estrogen, are pro-fibrotic in the skin. In the connective tissue disease, systemic sclerosis (SSc), both TGFβ and E2 are likely pathogenic. Yet the regulation of TGFβ in E2-induced dermal fibrosis remains ill-defined. Elucidating those regulatory mechanisms will improve the understanding of fibrotic disease pathogenesis and set the stage for developing potential therapeutics. Using E2-stimulated primary human dermal fibroblasts in vitro and human skin tissue ex vivo, we identified the important regulatory proteins for TGFβ and investigated the extracellular matrix (ECM) components that are directly stimulated by E2-induced TGFβ signaling. Methods We used primary human dermal fibroblasts in vitro and human skin tissue ex vivo stimulated with E2 or vehicle (ethanol) to measure TGFβ1 and TGFβ2 levels using quantitative PCR (qPCR). To identify the necessary cell signaling proteins in E2-induced TGFβ1 and TGFβ2 transcription, human dermal fibroblasts were pre-treated with an inhibitor of the extracellular signal-regulated kinase/mitogen-activated protein kinase (ERK/MAPK) pathway, U0126. Finally, human skin tissue ex vivo was pre-treated with SB-431542, a TGFβ receptor inhibitor, and ICI 182,780, an estrogen receptor α (ERα) inhibitor, to establish the effects of TGFβ and ERα signaling on E2-induced collagen 22A1 (Col22A1) transcription. Results We found that expression of TGFβ1, TGFβ2, and Col22A1, a TGFβ-responsive gene, is induced in response to E2 stimulation. Mechanistically, Col22A1 induction was blocked by SB-431542 and ICI 182,780 despite E2 stimulation. Additionally, inhibiting E2-induced ERK/MAPK activation and early growth response 1 (EGR1) transcription prevents the E2-induced increase in TGFβ1 and TGFβ2 transcription and translation. Conclusions We conclude that E2-induced dermal fibrosis occurs in part through induction of TGFβ1, 2, and Col22A1, which is regulated through EGR1 and the MAPK pathway. Thus, blocking estrogen signaling and/or production may be a novel therapeutic option in pro-fibrotic diseases.

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Caffeic Acid Phenethyl Ester Inhibits UV-Induced MMP-1 Expression by Targeting Histone Acetyltransferases in Human Skin

International Journal of Molecular Sciences ◽

10.3390/ijms20123055 ◽

2019 ◽

Vol 20 (12) ◽

pp. 3055 ◽

Cited By ~ 4

Author(s):

Eun Ju Shin ◽

Seongin Jo ◽

Hyo-kyoung Choi ◽

Sungbin Choi ◽

Sanguine Byun ◽

...

Keyword(s):

Caffeic Acid ◽

Human Skin ◽

Ex Vivo ◽

Bioactive Compound ◽

Caffeic Acid Phenethyl Ester ◽

Inhibitory Effect ◽

Dermal Fibroblasts ◽

Regulatory Pathways ◽

Skin Photoaging ◽

Hdf Cells

Caffeic acid phenethyl ester (CAPE), a naturally occurring bioactive compound, displays anti-inflammatory, anti-carcinogenic, and anti-microbial effects. However, the effect of CAPE on skin photoaging is unknown. Herein, we investigated the inhibitory effect of CAPE against ultraviolet (UV) irradiation-mediated matrix metalloproteinase (MMP)-1 expression and its underlying molecular mechanism. CAPE treatment suppressed UV-induced MMP-1 levels in both human dermal fibroblasts (HDF) and human skin tissues. While CAPE did not display any significant effects against the upstream regulatory pathways of MMP-1, CAPE was capable of reversing UV-mediated epigenetic modifications. CAPE suppressed UV-induced acetyl-histone H3 (Lys9) as well as total lysine acetylation in HDF cells. Similarly, CAPE also attenuated UV-induced lysine acetylations in human skin tissues, suggesting that the CAPE-mediated epigenetic alterations can be recapitulated in ex vivo conditions. CAPE was found to attenuate UV-induced histone acetyltransferase (HAT) activity in HDF. Notably, CAPE was able to directly inhibit the activity of several HATs including p300, CREP-binding protein (CBP), and p300/CBP-associated factor (PCAF), further confirming that CAPE can function as an epigenetic modulator. Thus, our study suggests that CAPE maybe a promising agent for the prevention of skin photoaging via targeting HATs.

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Pulsed Heat Shocks to Enhance Collagen Production of Human Dermal Fibroblasts in Ex-Vivo Skin

ASME 2010 Summer Bioengineering Conference, Parts A and B ◽

10.1115/sbc2010-19212 ◽

2010 ◽

Author(s):

S. D. Dams ◽

M. de Liefde ◽

A. M. Nuijs ◽

C. W. J. Oomens ◽

F. P. T. Baaijens

Keyword(s):

Ex Vivo ◽

Contemporary Literature ◽

Collagen Matrix ◽

Dermal Fibroblasts ◽

Human Dermal Fibroblasts ◽

Procollagen Type ◽

Different Temperatures ◽

Heat Shocks ◽

Ablative Techniques ◽

Type 3

Well-known characteristics of aging skin are the development of fine lines and wrinkles, but also changes in skin tone, skin texture, thickness and moisture content [1, 2]. Rejuvenation of the skin aims at reversing the signs of aging and can be established in the epidermis as well as in the dermis. Aged dermis for that matter has a degenerated collagen matrix. To regenerate this matrix, fibroblasts need to be stimulated into synthesizing new collagen. Among the rejuvenation methods used the non-ablative techniques are gaining popularity. These methods produce dermal remodeling without obvious epidermal injury using a thermal approach. The generation of heat would cause collagen injury and contraction followed by collagen synthesis [2, 3]. However, little research on physiological evidence can be found in contemporary literature. In this study, the effects of heat shocks of different temperatures on the expression of procollagen type 1and type 3 of human dermal fibroblasts in ex-vivo skin are investigated.

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Propolis Suppresses UV-Induced Photoaging in Human Skin through Directly Targeting Phosphoinositide 3-Kinase

Nutrients ◽

10.3390/nu12123790 ◽

2020 ◽

Vol 12 (12) ◽

pp. 3790

Author(s):

Da Hyun Kim ◽

Joong-Hyuck Auh ◽

Jeongyeon Oh ◽

Seungpyo Hong ◽

Sungbin Choi ◽

...

Keyword(s):

Protein Kinase ◽

Human Skin ◽

Therapeutic Potential ◽

Skin Aging ◽

Dermal Fibroblasts ◽

Kinase Assay ◽

Aging Effects ◽

Propolis Extract ◽

Pi3k Activity ◽

Phosphoinositide 3 Kinase

Propolis is a resinous substance generated by bees using materials from various plant sources. It has been known to exhibit diverse bioactivities including anti-oxidative, anti-microbial, anti-inflammatory, and anti-cancer effects. However, the direct molecular target of propolis and its therapeutic potential against skin aging in humans is not fully understood. Herein, we investigated the effect of propolis on ultraviolet (UV)-mediated skin aging and its underlying molecular mechanism. Propolis suppressed UV-induced matrix metalloproteinase (MMP)-1 production in human dermal fibroblasts. More importantly, propolis treatment reduced UV-induced MMP-1 expression and blocked collagen degradation in human skin tissues, suggesting that the anti-skin-aging activity of propolis can be recapitulated in clinically relevant conditions. While propolis treatment did not display any noticeable effects against extracellular signal-regulated kinase (ERK), p38, and c-jun N-terminal kinase (JNK) pathways, propolis exerted significant inhibitory activity specifically against phosphorylations of phosphoinositide-dependent protein kinase-1 (PDK1) and protein kinase B (Akt). Kinase assay results demonstrated that propolis can directly suppress phosphoinositide 3-kinase (PI3K) activity, with preferential selectivity towards PI3K with p110α and p110δ catalytic subunits over other kinases. The content of active compounds was quantified, and among the compounds identified from the propolis extract, caffeic acid phenethyl ester, quercetin, and apigenin were shown to attenuate PI3K activity. These results demonstrate that propolis shows anti-skin-aging effects through direct inhibition of PI3K activity.

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DNA macroarray study of skin aging-related genes expression modulation by antioxidant plant extracts on a replicative senescence model of human dermal fibroblasts

Phytotherapy Research ◽

10.1002/ptr.3308 ◽

2010 ◽

Vol 25 (5) ◽

pp. 686-693 ◽

Cited By ~ 5

Author(s):

Stéphanie Dudonné ◽

Philippe Coutière ◽

Marion Woillez ◽

Jean-Michel Mérillon ◽

Xavier Vitrac

Keyword(s):

Plant Extracts ◽

Replicative Senescence ◽

Skin Aging ◽

Dermal Fibroblasts ◽

Human Dermal Fibroblasts ◽

Genes Expression ◽

Expression Modulation

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(−)-Loliolide Isolated from Sargassum horneri Abate UVB-Induced Oxidative Damage in Human Dermal Fibroblasts and Subside ECM Degradation

Marine Drugs ◽

10.3390/md19080435 ◽

2021 ◽

Vol 19 (8) ◽

pp. 435

Author(s):

Ilekuttige Priyan Shanura Fernando ◽

Soo-Jin Heo ◽

Mawalle Kankanamge Hasitha Madhawa Dias ◽

Dissanayaka Mudiyanselage Dinesh Madusanka ◽

Eui-Jeong Han ◽

...

Keyword(s):

Inflammatory Responses ◽

Skin Aging ◽

Dermal Fibroblasts ◽

Sargassum Horneri ◽

Mitogen Activated Protein Kinase ◽

Ros Generation ◽

Protective Effects ◽

Human Dermal Fibroblasts ◽

Mitogen Activated Protein ◽

Uvb Exposure

Ultraviolet (UV) B exposure is a prominent cause of skin aging and a contemporary subject of interest. The effects are progressing through the generation of reactive oxygen species (ROS) that alter cell signaling pathways related to inflammatory responses. The present study evaluates the protective effects of (7aR)-6-hydroxy-4,4,7a-trimethyl-6,7-dihydro-5H-1-benzofuran-2-one (HTT) isolated from the edible brown algae Sargassum horneri against UVB protective effects in human dermal fibroblasts (HDFs). HTT treatment dose-dependently suppressed intracellular ROS generation in HDFs with an IC50 of 62.43 ± 3.22 µM. HTT abated UVB-induced mitochondrial hyperpolarization and apoptotic body formation. Furthermore, UVB-induced activation of key nuclear factor (NF)-κB and mitogen-activated protein kinase signaling proteins were suppressed in HTT treated cells while downregulating pro-inflammatory cytokines (interleukin-1β, 6, 8, 33 and tumor necrosis factor-α). Moreover, HTT treatment downregulated matrix metalloproteinase1, 2, 3, 8, 9 and 13 that was further confirmed by the inhibition of collagenase and elastase activity. The evidence implies that HTT delivers protective effects against premature skin aging caused by UVB exposure via suppressing inflammatory responses and degradation of extracellular matrix (ECM) components. Extensive research in this regard will raise perspectives for using HTT as an ingredient in UV protective ointments.

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Protective Effect of Polymethoxyflavones Isolated from Kaempferia parviflora against TNF-α-Induced Human Dermal Fibroblast Damage

Antioxidants ◽

10.3390/antiox10101609 ◽

2021 ◽

Vol 10 (10) ◽

pp. 1609

Author(s):

Hung Manh Phung ◽

Sullim Lee ◽

Sukyung Hong ◽

Sojung Lee ◽

Kiwon Jung ◽

...

Keyword(s):

Oxidative Stress ◽

Inflammatory Responses ◽

Skin Aging ◽

Dermal Fibroblasts ◽

Mitogen Activated Protein Kinase ◽

Type I ◽

Skin Damage ◽

Kaempferia Parviflora ◽

Matrix Metalloproteinase 1 ◽

Tnf Α

Similar to other organs, the skin undergoes a natural aging process. Moreover, constant direct exposure to environmental stresses, including ultraviolet irradiation, causes the signs of skin aging to appear rather early. Reactive oxygen species (ROS) and inflammatory responses accelerate skin damage in extrinsic aging. In this study, we aimed to investigate the skin protective effects of polymethoxyflavones found in Kaempferia parviflora against oxidative stress and inflammation-induced damage in human dermal fibroblasts (HDFs) stimulated by tumor necrosis factor-α (TNF-α). The experimental data identified 5,7,4′ trimethoxyflavone (TMF) as the most potent constituent in preventing TNF-α-induced HDF damage among the tested compounds and it was not only effective in inhibiting matrix metalloproteinase-1 (MMP-1) production but also in stimulating collagen, type I, and alpha 1 (COLIA1) expression. TMF suppressed TNF-α-stimulated generation of ROS and pro-inflammatory mediators, such as cyclooxygenase-2 (COX-2), interleukin (IL)-1β, and IL-6 in HDFs. TMF also inhibited the pathways regulating fibroblast damage, including mitogen-activated protein kinase (MAPK), activator protein 1 (AP-1), and nuclear factor-kappa B (NF-κB). In conclusion, TMF may be a potential agent for preventing skin aging and other dermatological disorders associated with oxidative stress and inflammation.

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