UVA-irradiated Human Dermal Fibroblasts Research Articles

Effects of rice fermentation and its bioactive components on UVA-induced oxidative stress and senescence in dermal fibroblasts.

Photoaging, caused by ultraviolet (UV) radiation, is characterized by the senescence of skin cells and reduction of collagens. Although rice fermentation is widely used in the cosmetics, its impact on skin photoaging is still not well understood. Herein, we investigated the possible effectiveness of Maifuyin, a fermented rice product, and its components, succinic acid (SA), and choline, for safeguarding UVA-exposed human dermal fibroblasts (HDFs) against photoaging. In this study, the effects of Maifuyin, SA, and choline on UVA-induced cell death and senescence in fibroblasts were evaluated in cell counting kit-8 (CCK-8), expression of β-galactosidase (β-GAL), and matrix metalloproteinases (MMP)-1. To identify oxidative stress, the investigation focused on reactive oxygen species, glutathione, superoxide dismutase, and malondialdehyde. Additionally, a mRNA sequencing technology (RNA-seq) was applied to study the underlying mechanisms of these components on UVA-induced photoaging. Meanwhile, the level of C-X-C motif chemokine ligand 2 (CXCL2) in the cell supernatant was confirmed to assess the autocrine chemokine level. To reassess the involvement of CXCL2, the expression of β-GAL was evaluated in fibroblasts treated with or without CXCL2. The results indicated that 1 mg/mL Maifuyin and SA inhibited UVA-induced senescence in fibroblasts, MMP-1 expression, and oxidative damage. The RNA-seq revealed 1 mg/mL Maifuyin and SA might be recruited chemokine CXCLs to inhibit MMPs production and fibroblast senescence via TNFα, MAPK, and NF-κB pathways. ELISA results showed a significant reduction of autocrine CXCL2 in UVA-irradiated HDFs by pretreating Maifuyin and SA. The β-GAL staining assay revealed that CXCL2 treatment increased β-GAL activity, while the administration of Maifuyin and SA counteracted this effect in HDFs. These results highlighted the potential use of Maifuyin and SA as promising candidates for anti-photoaging applications.

Anti-inflammatory, antioxidant and photoprotective activity of standardised Gaultheria procumbens L. leaf, stem, and fruit extracts in UVA-irradiated human dermal fibroblasts

Ethnopharmacological relevanceGaultheria procumbens L. is a polyphenolic-rich medicinal and food plant. Its leaves, stems, and fruits are traditional anti-inflammatory, antipyretic, antioxidant, and antimicrobial herbal medicines used to treat internal and external inflammation-related ailments, including rheumatic diseases, influenza, the common cold, fever, and skin and periodontal problems. Moreover, G. procumbens leaf extract is used for skin care as an anti-ageing and anti-wrinkle ingredient. Aim of the studyVarious environmental factors, especially solar ultraviolet radiation, accelerate skin ageing by promoting oxidative stress and inflammation. Despite the dermoprotective and anti-ageing applications, the impact of G. procumbens on human dermal fibroblasts is unknown. Therefore, the study aimed to evaluate the anti-inflammatory, antioxidant, and photoprotective activity of G. procumbens standardised leaf, stem, and fruit extracts in cellular models, including human dermal fibroblasts (Hs68 cells) under UVA-irradiation, the primary pro-ageing skin stressor. Materials and methodsHs68 fibroblasts were pre-treated (24h) with G. procumbens extracts (0.5–100 μg/mL) or reference compounds followed by UVA-irradiation (8 J/cm2). Cell viability and metabolic activity were measured by CCK-8 and MTT assays in human Hs68 and mouse L929 fibroblasts, respectively. The ROS level, SOD, and GST activities were estimated by fluorescence and spectrophotometric techniques. The pro-inflammatory potential (NF-κB transcription factor activation) was checked using THP1-Blue™ NF-κB cells, and the anti-inflammatory activity was studied by measuring IL-8, ICAM-1, and NF-κB levels and phosphorylation of Erk kinase in LPS-stimulated Hs68 cells by spectrophotometry and confocal microscopy. The UVA-induced DNA damage and cell migration were evaluated by comet and scratch assays, respectively. ResultsThe extracts did not affect the metabolic activity of mouse L929 fibroblasts and the viability of unirradiated human Hs68 cells. Additionally, the extracts noticeably enhanced the viability of UVA-irradiated Hs68 cells up to 115–120% (p < 0.001) for stem and leaf extract at 25 μg/mL. All extracts in a wide concentration range (0.5–100 μg/mL) did not activate monocytes or induce the NF-κB transcription factor in LPS-stimulated Hs68 fibroblasts. On the other hand, the extracts (5–25 μg/mL) restored the activity of endogenous antioxidant enzymes, i.e., SOD and GST, up to 120–140% (p < 0.001) in the UVA-irradiated Hs68 cells. Moreover, a statistically significant reduction of ROS, IL-8, ICAM-1, and NF-κB levels by up to 48%, 88%, 43%, and 39%, respectively (p < 0.001) and strong suppression of Erk kinase activation was observed for the extracts (25–50 μg/mL) in LPS-stimulated human fibroblasts. The total DNA damage (% tail DNA) in irradiated Hs68 cells was also strongly decreased by up to 66–69% (p < 0.001) at 50 μg/mL. However, the treatment with the extracts did not relevantly enhance the cell migration of Hs68 fibroblasts. ConclusionsThe results suggest that G. procumbens may effectively protect human skin fibroblast from UVA irradiation. The leaf and stem extracts were the most potent antioxidants, while fruit and stem extracts revealed the strongest anti-inflammatory activity. The observed effects support the traditional use of aerial plant parts (leaves, stems, and fruits) in treating inflammation-related skin disorders cross-linked with oxidative stress and the topical application of Gaultheria extracts as anti-ageing agents intended for skin care.

Anti-aging Effect of Sargassum patens Extract in UVA-irradiated Human Dermal Fibroblasts

Purpose: Ultraviolet (UV) irradiation causes extrinsic aging of human skin. It is believed that UV irradiation caused aging phenomena such as skin erythema, wrinkle formation, and skin elasticity reduction.Methods: The effect of Sargassum patens extract on COL1A1 and MMP1 production in normal human dermal fibroblasts was investigated as a potential cosmetic raw material.Results: In this study, I assessed how Sargassum patens extract affected COL1A1/COL3A1/MMP1/SODs/Catalase mRNA expression and protein production of Collagen, type I and MMP1. Quantitative real-time reverse transcription–polymerase chain reaction demonstrated that the Sargassum patens extract increased the mRNA levels of COL1A1/COL3A1/SOD2/CAT genes. Additionally, when compared to UVA-treated dermal fibroblasts, collagen and type I protein increased in Sargassum patens extract-treated conditions. Furthermore, MMP1 mRNA and secreted MMP1 protein expression were reduced by Sargassum patens extract treatment. According to these findings, Sargassum patens extract increased antiaging and antioxidative effects in dermal fibroblasts.Conclusion: Therefore, I identified the antiaging effects of Sargassum patens extract, and these findings indicated that Sargassum patens could be a pharmaceutical ingredient for skin antiaging. Based on this, I expected to conduct additional research on Sargassum patens extract for cellular mechanisms.

Chlorogenic Acid Prevents UVA-Induced Skin Photoaging through Regulating Collagen Metabolism and Apoptosis in Human Dermal Fibroblasts.

Skin aging is categorized as chronological aging and photo-aging that affected by intrinsic and extrinsic factors. The present study aimed to investigate the anti-aging ability and its underlying mechanism of chlorogenic acid (CGA) on human dermal fibroblasts (HDFs). In this study, CGA specifically up-regulated collagen I (Col1) mRNA and protein expressions and increased the collagen secretion in the supernatant of HDFs without affecting the cell viability, the latter was also demonstrated in BioMAP HDF3CGF system. Under ultraviolet A (UVA)-induced photoaging, CGA regulated collagen metabolism by increasing Col1 expression and decreasing matrix metalloproteinase 1 (MMP1) and MMP3 levels in UVA-irradiated HDFs. The activation of transforming growth factor-β (TGF-β)-mediated Smad2/3 molecules, which is crucial in Col1 synthesis, was suppressed by UVA irradiation and but enhanced at the presence of CGA. In addition, CGA reduced the accumulation of UVA-induced reactive oxygen species (ROS), attenuated the DNA damage and promoted cell repair, resulting in reducing the apoptosis of UVA-irradiated HDFs. In conclusion, our study, for the first time, demonstrate that CGA has protective effects during skin photoaging, especially triggered by UVA-irradiation, and provide rationales for further investigation of CGA being used to prevent or treat skin aging.

Comparison Between Injectable Platelet-rich Fibrin and Platelet-rich Plasma in Ameliorating UVA-induced Photoaging in Human Dermal Fibroblasts via the Activation of TGF-β/Smad Signaling Pathway.

Early-stage photoaging is characterized by skin laxity and wrinkling, which are mainly attributable to the ultraviolet (UV) irradiation-mediated imbalance between matrix metalloproteinase (MMP) production and collagen degradation. Injectable platelet-rich fibrin (i-PRF) is a novel blood concentrate with potential effects on photoaging. Over the past few decades, platelet-rich plasma (PRP) has been widely researched and used in different clinical fields as a first-generation platelet concentrate. The aim of this study was to compare the antiphotoaging effects of i-PRF in UVA-irradiated human dermal fibroblasts with those of PRP by examining cell proliferation, migration and apoptosis, ROS generation, MMP-1 and collagen I levels. The activation of the TGF-β/Smad signaling pathway by i-PRF and PRP was also investigated using western blotting. The results showed that i-PRF was more effective than PRP in promoting cell proliferation and migration. Moreover, i-PRF reduced ROS generation and cell apoptosis more effectively than PRP. With respect to the mechanism of collagen I upregulation, stronger stimulation of the TGF-β/Smad signaling pathway and greater suppression of MMP-1 expression were achieved by i-PRF than by PRP. Our results suggest that i-PRF can be a promising substitute for PRP in alleviating UVA-induced photoaging and should be explored further for its anti-photoaging properties.

A Negative Feedback Loop in Ultraviolet A-Induced Senescence in Human Dermal Fibroblasts Formed by SPCA1 and MAPK.

Secretory pathway calcium ATPase 1 (SPCA1) is a calcium pump localized specifically to the Golgi. Its effects on UVA-induced senescence have never been examined. In our study, expression of SPCA1 was increased in UVA-irradiated human dermal fibroblasts (HDFs) by activating mitogen-activated protein kinase (MAPK) and its downstream transcription factor, c-jun. Dual-luciferase reporter and chromatin immunoprecipitation assays revealed that c-jun regulated SPCA1 by binding to its promoter. Furthermore, downregulating SPCA1 with siRNA transfection aggravated UVA-induced senescence due to an elevation of intracellular calcium concentrations and a subsequent increase in reactive oxygen species (ROS) and MAPK activity. In contrast, overexpression of SPCA1 reduced calcium overload, consequently lowering the ROS level and suppressing MAPK activation. This alleviated the cellular senescence caused by UVA irradiation. These results indicated that SPCA1 might exert a protective effect on UVA-induced senescence in HDFs via forming a negative feedback loop. Specifically, activation of MAPK/c-jun triggered by UVA transcriptionally upregulated SPCA1. In turn, the increased SPCA1 lowered the intracellular Ca2+ level, probably through pumping Ca2+ into the Golgi, leading to a reduction of ROS, eventually decreasing MAPK activity and diminishing UVA-induced senescence.

Santamarine Shows Anti-Photoaging Properties via Inhibition of MAPK/AP-1 and Stimulation of TGF-β/Smad Signaling in UVA-Irradiated HDFs.

Chronic UVA exposure results in elevated reactive oxygen species in skin which leads to photoaging characterized as upregulated matrix metalloproteinase (MMP)-1 and loss of collagen. Therefore, natural antioxidants are hailed as promising agents to be utilized against photoaging. In the current study, reynosin and santamarine, two known sesquiterpene lactones isolated from Artemisia scoparia, were analyzed for their anti-photoaging properties in UVA-irradiated human dermal fibroblasts (HDFs). Results showed that UVA irradiation (8 J/cm2) upregulated the MMP-1 secretion and expression, and suppressed collagen production, which were significantly reverted by santamarine treatment (10 µM). Although both reynosin and santamarine exhibited ROS scavenging abilities, reynosin failed to significantly diminish UVA-stimulated MMP-1 release. UVA-irradiated HDFs showed increased collagen production when treated with santamarine. As a mechanism to suppress MMP-1, santamarine significantly suppressed the UVA-induced phosphorylation of p38 and JNK and nuclear translocation of p-c-Fos and p-c-Jun. Santamarine promoted collagen I production via relieving the UVA-induced suppression on TGF-β and its downstream activator Smad2/3 complex. Antioxidant properties of santamarine were also shown to arise from stimulating Nrf2-dependent expression of antioxidant enzymes SOD-1 and HO-1 in UVA-irradiated HDFs. In conclusion, santamarine was found to be a promising natural antioxidant with anti-photoaging properties against UVA-induced damages in HDFs.

Antiphotoaging Effect of 3,5-Dicaffeoyl-epi-quinic Acid against UVA-Induced Skin Damage by Protecting Human Dermal Fibroblasts In Vitro.

Cutaneous aging is divided into intrinsic and exogenous aging correspondingly contributing to the complex biological phenomenon in skin. Intrinsic aging is also termed chronological aging, which is the accumulation of inevitable changes over time and is largely genetically determined. Superimposed on this intrinsic process, exogenous aging is associated with environmental exposure, mainly to ultraviolet (UV) radiation and more commonly termed as photoaging. UV-induced skin aging induces increased expression of matrix metalloproteinases (MMPs) which in turn causes the collagen degradation. Therefore, MMP inhibitors of natural origin are regarded as a primary approach to prevent or treat photoaging. This study investigated the effects of 3,5-dicaffeoyl-epi-quinic acid (DEQA) on photoaging and elucidated its molecular mechanisms in UVA-irradiated human dermal fibroblasts (HDFs). The results show that treatment with DEQA decreases MMP-1 production and increases type I collagen production in UVA-damaged HDFs. In addition, treatment of UVA-irradiated HDFs with DEQA downregulates MMP-1, MMP-3 and MMP-9 expression via blocking MAPK-cascade-regulated AP-1 transcriptional activity in UVA-irradiated HDFs. Furthermore, DEQA relieves the UVA-mediated suppression of type I procollagen and collagen expression through stimulating TGF-β/Smad signaling, leading to activation of the Smad 2/3 and Smad 4 nuclear translocation. These results suggest that DEQA could be a potential cosmetic agent for prevention and treatment of skin photoaging.

Enzyme-Treated Zizania latifolia Ethanol Extract Protects from UVA Irradiation-Induced Wrinkle Formation via Inhibition of Lysosome Exocytosis and Reactive Oxygen Species Generation.

Ultraviolet A (UVA) is a risk factor for photoaging and wrinkle formation. Zizania latifolia is an herbaceous perennial plant. It contains many bioactive compounds such as tricin that show antioxidative and anti-inflammatory effects. The aim of this study was to investigate the antiwrinkle effect of a mixture of hydrolytic enzyme (cellulase, hemicellulase and pectinase)-treated Z. latifolia extract (ZLE) and tricin on UVA-irradiated human dermal fibroblasts (HDFs) and SKH-1 hairless mice. Treatment of UVA-irradiated HDF cells with ZLE and tricin significantly decreased UVA induced-plasma membrane rupture, generation of ROS, expression levels of total and secreted lysosomal associated membrane protein (LAMP-1), cathepsin B and metalloproteinases (MMPs) and inhibited NF-κB activation. In the animal study, UVA-damaged epidermal and dermal tissues were repaired by the ZLE and tricin treatments. Administration of ZLE or tricin to UVA-irradiated animals recovered skin surface moisture and collagen fiber in dermal tissue. Treatment of ZLE or tricin decreased wrinkle formation, secretion of MMPs and expression levels of vascular endothelial growth factor (VEGF) and cathepsin B, and increased the expression level of collagen-1 in UVA-irradiated animals. Overall, the ZLE and tricin treatments decreased the skin damage induced by UVA irradiation via inhibition of lysosomal exocytosis and ROS generation. Therefore, ZLE and tricin are promising as antiwrinkle and antiphotoaging agents.

Gold Nanoparticles Using Ecklonia stolonifera Protect Human Dermal Fibroblasts from UVA-Induced Senescence through Inhibiting MMP-1 and MMP-3.

The effect of gold nanoparticles (GNPs) synthesized in marine algae has been described in the context of skin, where they have shown potential benefit. Ecklonia stolonifera (ES) is a brown algae that belongs to the Laminariaceae family, and is widely used as a component of food and medicine due to its biological activities. However, the role of GNPs underlying cellular senescence in the protection of Ecklonia stolonifera gold nanoparticles (ES-GNPs) against UVA irradiation is less well known. Here, we investigate the antisenescence effect of ES-GNPs and the underlying mechanism in UVA-irradiated human dermal fibroblasts (HDFs). The DPPH and ABTS radical scavenging activity of ES extracts was analyzed. These analyses showed that ES extract has potent antioxidant properties. The facile and optimum synthesis of ES-GNPs was established using UV-vis spectra. The surface morphology and crystallinity of ES-GNPs were demonstrated using high resolution transmission electron microscopy (HR-TEM), energy dispersive spectroscopy (EDS), X-ray diffraction (XRD), and Fourier-transform infrared spectroscopy (FT-IR). ES-GNPs presented excellent photocatalytic activity, as shown by the photo-degradation of methylene blue and rhodamine B. A cellular senescence model was established by irradiating HDFs with UVA. UVA-irradiated HDFs exhibited increased expression of senescence-associated β-galactosidase (SA-β-galactosidase). However, pretreatment with ES-GNPs resulted in reduced SA-β-galactosidase activity in UVA-irradiated HDFs. Intracellular ROS levels and G1 arrest in UVA-irradiated HDFs were checked against the background of ES-GNP treatment to investigate the antisenescence effects of ES-GNPs. The results showed that ES-GNPs significantly inhibit UVA-induced ROS levels and G1 arrest. Importantly, ES-GNPs significantly downregulated the transcription and translation of MMP (matrix metalloproteinases)-1/-3, which regulate cellular senescence in UVA-irradiated HDFs. These findings indicate that our optimal ES-GNPs exerted an antisenescence effect on UVA-irradiated HDFs by inhibiting MMP-1/-3 expression. Collectively, we posit that ES-GNPs may potentially be used to treat photoaging of the skin.

Antiphotoaging Effect of (2′S)-Columbianetin from Corydalis heterocarpa in UVA-Irradiated Human Dermal Fibroblasts

Ultraviolet A (UVA)-induced detrimental effects in the skin, also known as photoaging, are mediated with several pathways including oxidative stress generation and extracellular matrix (ECM) degradation. UVA irradiation results in excessive production of matrix metalloproteinases (MMPs), enzymes responsible for the degradation of ECM components such as collagen. In this study, the protective effects of (2′S)-columbianetin against UVA-induced changes in matrix metalloproteinase-1 (MMP-1) and collagen production were investigated in human dermal fibroblasts (HDFs). The (2′S)-columbianetin was isolated from Corydalis heterocarpa. UVA exposure increased MMP-1 release from HDFs and diminished the release of type I pro-collagen. Treatment with (2′S)-columbianetin reversed these effects of UVA exposure. The (2′S)-columbianetin treatment also suppressed the intracellular expression of MMP-1 and increased type I pro-collagen expression. UVA exposure elevated the activation of p38, c-Jun-amino-terminal kinase (JNK) and extracellular signal-related kinase (ERK) as the mechanism to stimulate MMP-1 production. The (2′S)-columbianetin suppressed the phosphorylation of JNK and ERK. The (2′S)-columbianetin was also stimulated collagen production via TGFβ signaling cascade, relieving UVA-induced suppression of Smad2/3 phosphorylation and translocation. In conclusion, (2′S)-columbianetin was suggested to be a potential cosmeceutical lead compound with antiphotoaging properties against UVA-induced collagen degradation.

Anticatabolic and Anti-Inflammatory Effects of Myricetin 3-O-β-d-Galactopyranoside in UVA-Irradiated Dermal Cells via Repression of MAPK/AP-1 and Activation of TGFβ/Smad.

UV irradiation is one of the main causes of extrinsic skin aging. UV-mediated skin aging, also known as photoaging, causes excessive breakdown of extracellular matrix which leads skin to lose its elasticity and strength. Several phytochemicals are known to exert anti-photoaging effects via different mechanisms, partly due to their antioxidant properties. The current study has been carried out to determine the potential anti-photoaging properties of myricetin 3-O-β-d-galacto-pyranoside (M3G), a flavonol glycoside isolated from L. tetragonum, in UVA-irradiated in vitro models; HaCaT keratinocytes and human dermal fibroblasts (HDFs). UVA-induced changes in MMP-1 and collagen production have been observed in HaCaT keratinocytes and HDFs. Further, UVA-induced activation of MAPK signaling, and pro-inflammatory cytokine production have been investigated. TGFβ/Smad pathway has also been analyzed in UVA-irradiated HDFs. Treatment with M3G reversed the UVA-induced changes in MMP-1 and collagen production both in HaCaT keratinocytes and HDFs. UVA-mediated activation of p38, ERK and JNK MAPK activation was also inhibited by M3G treatment in HaCaT keratinocytes. In HDFs, M3G was able to upregulate the TGFβ/Smad pathway activation. In addition, M3G downregulated the UVA-induced pro-inflammatory cytokines in keratinocytes and HDFs. It has been suggested that the M3G has exerted potential antiphotoaging properties in vitro, by attenuating UVA-induced changes in MMP-1 and collagen production in keratinocytes and dermal fibroblasts.

The Antiaging Activity of Ergothioneine in UVA-Irradiated Human Dermal Fibroblasts via the Inhibition of the AP-1 Pathway and the Activation of Nrf2-Mediated Antioxidant Genes

UVA irradiation induced ROS-mediated photo damage to the human skin leading to coarseness, wrinkling, pigmentation, and cutaneous malignancies. We investigated the dermatoprotective efficacies of submicromolar concentrations of ergothioneine (EGT, 0.125-0.5 μM), which occurs naturally as a sulfur-containing amino acid, in the mechanisms in human skin fibroblast (HSF) cells. UVA-induced AP-1 (c-Fos and c-Jun) translocation was found to be inhibited by EGT treatments with the parallel inhibition of the collagenolytic matrix metalloproteinase- (MMP-) 1 activation and type I procollagen degradation. Moreover, EGT mitigated UVA-induced ROS generation. An increase in the amount of antioxidant genes (HO-1, NQO-1, and γ-GCLC) from EGT and were associated with upregulated Nrf2 expressions in a dose-dependent or time-dependent manner. We confirmed this from Nrf2 translocation and increased nuclear ARE promoter activity that underlie EGT dermatoprotective activities. Also, glutathione (GSH) levels (from γ-GCLC) were significantly increased. Moreover, we showed that mediated by ERK, JNK, and PKC, signaling cascades mediate Nrf2 translocation. We confirmed this phenomenon by the suppressed nuclear Nrf2 activation in cells that were treated with respective inhibitors (PD98059, SP600125, and GF109203X). However, antioxidant protein expressions were impaired in Nrf2 knockdown cells to confirm that ARE/Nrf2 pathways and the inhibition of AP-1 had significant roles in EGT-mediated protective effects. We can conclude that ergothioneine ameliorated UVA-induced skin aging and is a useful food supplement for skin care products.

Stachys riederi var. japonica Extract Reduces Cytochrome C Release from Mitochondria in UVA-irradiated Human Dermal Fibroblasts

This study was performed to investigate the cytoprotective effects of Stachys riederi var. japonica ethanol extract (SREE) to control oxidative stress induced by UVA-irradiation by examining antioxidant capacity and gene expression of cytochrome c using human dermal fibroblasts. The total polyphenol...

CircCOL3A1-859267 regulates type I collagen expression by sponging miR-29c in human dermal fibroblasts

The genes COL1A1 and COL1A2 encode the pro-alpha1 and pro-alpha2 chains of type I collagen, respectively, which is one of the main components of skin dermis. We have previously demonstrated that a circular RNA (a class of recently identified non-coding RNAs that exhibit regulatory potency by sequestering miRNAs like a sponge), which we termed circCOL3A1-859267, is downregulated and regulates type I collagen expression in UVA-exposed human dermal fibroblasts (HDFs). However, the precise mechanisms of circCOL3A1-859267-mediated collagen expression in UVA-irradiated HDFs remain unclear. To elucidate the mechanism of circCOL3A1-859267-mediated regulation of type I collagen expression. We initially predicted miRNA binding sites on circCOL3A1-859267 based on a bioinformatic method, and a dual luciferase reporter assay was used to determine miRNA binding to circCOL3A1-859267 in HEK 293 cells. The effect of UVA irradiation on the expression of miRNAs as well as circCOL3A1-859267-mediated type I collagen expression was further investigated in HDFs. miR-29a, miR-29b, miR-29c, miR-767, and miR-133a were predicted to bind both circCOL3A1-859267 and COL1A1/COL1A2, however, only miR-29c was shown to bind to circCOL3A1-859267 based on the dual luciferase reporter assay. In UVA-exposed HDFs, only miR-29c was upregulated. Finally, transfection of a small interfering RNA targeting circCOL3A1-859267 or miR-29c mimic suppressed the expression of type I collagen in HDFs; the miR-29c mimic-induced down-regulation was restored via overexpression of circCOL3A1-859267 using a lentiviral-based expression system. Our results indicate that circCOL3A1-859267 regulates type I collagen expression by sponging and sequestering miR-29c in HDFs.

Regenerative Effects of Wharton’s Jelly Stem Cells-Conditioned Medium in UVA-Irradiated Human Dermal Fibroblasts

Background: Ultraviolet A radiation (UVA) can photo-age skin by suppressing the proliferation, migration, and collagen deposition of human dermal fibroblasts (HDFs). This process occurs because UVA light can inhibit the gene expression of the TGF-β receptor in HDFs. Moreover, Wharton’s Jelly Stem Cells-Conditioned Medium (WJSC-CM) is hypothesized to release microvesicles that contain short m-RNA with regenerative properties.&#x0D; Objectives: This study aimed to determine the regenerative properties of WJSC-CM on UVA-Irradiated Human Dermal Fibroblasts (UVA-HDFs)&#x0D; Methods: Passaged fourth of HDFs obtained from the foreskin of six (11- to 13-year-old) boys were repeatedly irradiated with a total of 10 J/cm2 UVA and treated with various concentrations of WJSC-CM. We used non-irradiated HDFs as positive control. After that, the consumption of TGF-β, cellular proliferation, cellular migration, and collagen deposition of each group were measured and compared.&#x0D; Results: Compared to the non-irradiated groups, the proliferation rates, migration rates, and collagen deposition of UVA-HDFs significantly decreased (p&lt;0.05). WJSC-CM can improve the consumption of TGF-β, proliferation, and cellular migration of UVA-HDFs. However, WJSC-CM failed to improve the collagen deposition of UVA-HDFs (p&gt;0.05).&#x0D; Conclusions: WJSC-CM has regenerative properties and is a candidate material for the treatment of prematurely ageing skin induced by UVA-irradiation.&#x0D; &#x0D; Ethical permission: This experiment was permitted by the local ethical permission committee Ref:KE/FK/382/EC with permission letter dated 17-04-2013.

Anti-aging Effects of P7C3 in UVA-irradiated Human Dermal Fibroblasts

ëª©ì  ìžì™¸ì„ ì€ 인간의 피부에 ê´‘ë ¸í™”ë¥¼ ìœ ë°œí•˜ì—¬, 피부 íƒ„ë ¥ 감소, 주름 형성 및 상처 ì¹˜ìœ ì§€ì—°ê³¼ 같은 í”¼ë¶€ë ¸í™”í˜„ìƒì„ ì•¼ê¸°í•œë‹¤ê³ ì•Œë ¤ì ¸ 있다. ì‹ ê²½ë³´í˜¸ì œ 후보물질로 ì•Œë ¤ì§„ P7C3의 화장품 원료로서의 가능성을 확인하기 위하여 얼굴 ìœ ëž˜ 인간 ì„¬ìœ ì•„ì„¸í¬ì—ì„œ P7C3에 의한 collagen, type I, alpha 1 (COL1A1)ê³¼ matrix metallopeptidase 1 (MMP1)의 발현양상과 ìƒì²˜ì¹˜ìœ ë¶„ì„ë²•ì„ 수행하였다. 방법 Cell counting kit-8 (CCK-8) 분석법을 통해 ì„¸í¬ë ì„±í‰ê°€ì™€ 이후 실험에 ì‚¬ìš©ë ë†ë„ë¥¼ ì •í•˜ì˜€ë‹¤. Real-time polymerase chain reaction (real-time PCR)을 이용하여 COL1A1ê³¼ MMP1의 mRNA 발현양상을 확인하였으며, P7C3의 ìƒì²˜ì¹˜ìœ ëŠ¥ì„ wound healing assay를 통해 확인하였다. ê²°ê³¼ ìžì™¸ì„ ì— 의해 발현이 감소된 COL1A1이 P7C3에 의해 ì¦ê°€ë˜ì—ˆê³ , ìžì™¸ì„ ì— 의해 발현이 증가한 MMP1은 감소되었다. 더욱이 ìƒì²˜ì¹˜ìœ ëŠ¥ 실험을 통해 P7C3가 ì„¬ìœ ì•„ì„¸í¬ì˜ ì´ë™ëŠ¥ë ¥ì„ 회복시켜 ìƒì²˜ì¹˜ìœ ì— 도움이 ë ìˆ˜ 있음을 확인하였다. ê²°ë¡ ë³¸ 연구결과를 통하여 P7C3가 ë ¸í™”ëœ 피부를 ê°œì„ í• ìˆ˜ 있음을 í™•ì¸í•˜ì˜€ê³ , 향후P7C3가 기능성 화장품 개발에 ì‘ìš©ë ìˆ˜ 있는 소재로서의 가능성을 확인하기 위하여 ì¶”ê°€ì ì¸ 연구가 í•„ìš”í• ê²ƒì´ë‹¤. 핵심용어: P7C3, ìžì™¸ì„ , 콜라겐, Matrix metallopeptidase 1, ë ¸í™”

199 EFFECTS OF REPROGRAMMING-CONDITIONED MEDIUM ON ULTRAVIOLET RAY A–DAMAGED HUMAN DERMAL FIBROBLASTS

Ultraviolet ray A (UVA) is an electromagnetic light with a long wavelength from the sun. The penetration of UVA deep into the human dermis causes changes in cells, such as DNA fragmentation, apoptosis, and senescence, eventually leading a decline of proliferation and wound-healing ability. These changes induced by UVA exposure are similar to those seen in the process of stem cell differentiation. We postulated that the condition that reverses cellular differentiation may alleviate the UVA-induced damage in skin cells. Human dermal fibroblasts (HDF) could be reprogrammed to induced pluripotent stem cells (iPSC). Conditioned medium (CM) was prepared during the process of iPSC reprogramming (referred to as Repro-CM). The UVA-irradiated HDF were cultured in Repro-CM for 24 h. In comparison with CM prepared from the culture of normal HDF and iPSC (referred to as HDF-CM and iPSC-CM, respectively), effects of Repro-CM on UVA-irradiated cells were investigated. Viability, wound-healing ability, apoptosis, and senescence of HDF were analysed by WST-1 assay, scratch assay, Annexin V assay, and senescence-associated β-galactosidase assay, respectively. Upon recovering from the UVA-induced damage, viability and wound-healing ability of HDF were significantly different (P &lt; 0.05) among the treatments in the order of Repro-, HDF-, and iPSC-CM. In the same context, apoptosis and senescence were significantly different (P &lt; 0.05) in the order of iPSC-, HDF-, and Repro-CM. Interestingly, iPSC-CM did not substantially ameliorate UVA-induced damage, suggesting that the conditions optimized to pluripotent stem cells may not be suitable for the recovery from damage in terminally differentiated cells, such as fibroblasts. The RNA-seq analysis was performed to assess the genome-wide transcriptional profile in the process of recovery. Repro- and HDF-CM were categorized more closely than iPSC-CM in hierarchical cluster analysis. In comparison with iPSC-CM, the up-regulated genes by Repro-CM treatment were related to regulation of cell proliferation and cell metabolism, whereas down-regulated genes were related to antiapoptosis and response to stimulation of chemical and organic substances. Overall, providing an environment of reprogramming, as shown by Repro-CM in the present study, may assist recovery of HDF from UVA-induced damage. The results of the study may be applicable in developing pharmaceuticals to treat aging and wrinkling of the skin caused by UVA irradiation.

Licoricidin, an isoflavonoid isolated from Glycyrrhiza uralensis Fisher, prevents UVA-induced photoaging of human dermal fibroblasts.

Licoricidin is an isoflavonoid isolated from Glycyrrhiza uralensis Fisher. In this study, we investigated the effects of licoricidin on photoaging of UVA-irradiated human dermal fibroblasts (HDFs). In vitro reactive oxygen species (ROS) scavenging activity, cellular protective effect and inhibition of elastase activity was determined by Fe3+ -EDTA/H2 O2 systems, photohaemolysis and elastase activity assay, respectively. Anti-oxidative capacity of the compound was evaluated by fluorescent ELISA and 2', 7'-dichlorofluorescin-diacetate (DCF-DA) assay. The expression of protein and phosphorylation was examined using Western blot. The ROS scavenging activity (OSC50 ) of licoricidin was 2.77 μM. It was 3.1-fold higher than that of L-ascorbic acid. Its protective effects were confirmed in a study of 1 O2 -induced cellular damage to human erythrocytes. The τ50 value of 10 μM of licoricidin was 71.0 min; this was markedly higher than that obtained with α-tocopherol (37.0 min). The elastase inhibitory activity of licoricidin (IC50 of 61.2 μM) was 2.1-fold more potent than that of oleanolic acid. Licoricidin markedly reduced the UVA-induced intracellular ROS in a concentration-dependent manner. Western blot revealed that licoricidin attenuated the UVA-dependent induction of MMP-1 protein. Mechanistically, this appeared to be due to licoricidin-dependent inhibition of mitogen-activated protein kinases (MAPK) phosphorylation, which resulted in decreased c-Jun activation and reduced c-Jun and c-Fos expression. Licoricidin blocks UVA-induced photoaging via ROS scavenging. This activity converges to limit the activity of MMP-1. These data suggest that licoricidin may be considered as an active ingredient in new topically applied anti-ageing formulations.

Platelet-Rich Fibrin Lysate Can Ameliorate Dysfunction of Chronically UVA-Irradiated Human Dermal Fibroblasts

To determine whether platelet-rich fibrin lysate (PRF-L) could restore the function of chronically ultraviolet-A (UVA)-irradiated human dermal fibroblasts (HDFs), we isolated and sub-cultured HDFs from six different human foreskins. HDFs were divided into two groups: those that received chronic UVA irradiation (total dosages of 10 J cm-2) and those that were not irradiated. We compared the proliferation rates, collagen deposition, and migration rates between the groups and between chronically UVA-irradiated HDFs in control and PRF-L-treated media. Our experiment showed that chronic UVA irradiation significantly decreased (p<0.05) the proliferation rates, migration rates, and collagen deposition of HDFs, compared to controls. Compared to control media, chronically UVA-irradiated HDFs in 50% PRF-L had significantly increased proliferation rates, migration rates, and collagen deposition (p<0.05), and the migration rates and collagen deposition of chronically UVA-irradiated HDFs in 50% PRF-L were equal to those of normal fibroblasts. Based on this experiment, we concluded that PRF-L is a good candidate material for treating UVA-induced photoaging of skin, although the best method for its clinical application remains to be determined.