scholarly journals Up-regulation of tight junction-related proteins and increase of human epidermal keratinocytes barrier function by Saccharomycosis ferment filtrate

2011 ◽  
Vol 01 (01) ◽  
pp. 15-24
Author(s):  
Jong-Hwei Pang ◽  
Wen-Rou Wong ◽  
Tomohiro Hakozaki ◽  
Takashi Yoshii ◽  
Tzu-Ya Chen
Keyword(s):  
Tight Junction ◽  
Barrier Function ◽  
Epidermal Keratinocytes ◽  
Human Epidermal Keratinocytes ◽  
Related Proteins

scholarly journals Promoting effect of lactoferrin on barrier function and epithelial differentiation of human keratinocytes

2017 ◽  
Vol 95 (1) ◽  
pp. 64-68 ◽  
Author(s):  
Ryo Uchida ◽  
Reiji Aoki ◽  
Ayako Aoki-Yoshida ◽  
Atsushi Tajima ◽  
Yoshiharu Takayama
Keyword(s):  
P38 Mapk ◽  
Barrier Function ◽  
Regulatory Element ◽  
Human Keratinocytes ◽  
Epithelial Differentiation ◽  
Mitogen Activated Protein Kinase ◽  
Keratinocyte Differentiation ◽  
Epidermal Keratinocytes ◽  
Bovine Lactoferrin ◽  
Human Epidermal Keratinocytes

The purpose of this study was to elucidate the effects of bovine lactoferrin on keratinocyte differentiation and barrier function. Addition of bovine lactoferrin to differentiating HaCaT human keratinocytes led to increased transepithelial electrical resistance (TER), a marker of epithelial barrier function. This elevation was followed by upregulation of two differentiation markers, involucrin and filaggrin. The expression level of sterol regulatory element-binding protein-1 was also enhanced by bovine lactoferrin. The lactoferrin-induced upregulation of involucrin and filaggrin expression were confirmed in normal human epidermal keratinocytes (NHEK). Treatment with SB203580, a p38 mitogen-activated protein kinase (MAPK) α inhibitor, impaired the upregulation of involucrin and filaggrin expression in response to lactoferrin. The elevation of p38 MAPK phosphorylation was further enhanced by lactoferrin in the initial stage of differentiation of HaCaT keratinocytes. The findings suggest that bovine lactoferrin promotes epithelial differentiation by a p38-MAPK-dependent mechanism.

Activation of TRPV4 Strengthens the Tight-Junction Barrier in Human Epidermal Keratinocytes

2013 ◽  
Vol 26 (1) ◽  
pp. 15-21 ◽  
Author(s):  
Y. Akazawa ◽  
T. Yuki ◽  
H. Yoshida ◽  
Y. Sugiyama ◽  
S. Inoue
Keyword(s):  
Tight Junction ◽  
Epidermal Keratinocytes ◽  
Human Epidermal Keratinocytes

scholarly journals Filaggrin and tight junction proteins are crucial for IL-13-mediated esophageal barrier dysfunction

2018 ◽  
Vol 315 (3) ◽  
pp. G341-G350 ◽  
Author(s):  
Liping Wu ◽  
Tadayuki Oshima ◽  
Min Li ◽  
Toshihiko Tomita ◽  
Hirokazu Fukui ◽  
...  
Keyword(s):  
Epithelial Cells ◽  
Tight Junction ◽  
Barrier Function ◽  
Epithelial Barrier ◽  
Tight Junction Proteins ◽  
Barrier Dysfunction ◽  
Esophageal Epithelial Cells ◽  
Air Liquid Interface ◽  
Related Proteins ◽  
Junction Proteins

Eosinophilic esophagitis (EoE) is an allergy-mediated disease that is accompanied by IL-13 overexpression and an impaired esophageal barrier. Filaggrin (FLG) and tight junction (TJ) proteins are considered to contribute to epithelial barrier function. However, their functional involvement in EoE has not been elucidated. Here, we aimed to determine the IL-13-mediated barrier dysfunction and expression of TJ-related proteins in EoE and to characterize interactions among TJ-related proteins involved in the barrier function of the esophageal epithelium. Biopsy specimens from EoE patients were analyzed. Primary human esophageal epithelial cells (HEECs) were cultured using an air-liquid interface (ALI) system. The permeability of TJs was assayed by biotinylation. Transepithelial electrical resistance (TEER) was measured after stimulation with IL-13 and after siRNA silencing of FLG expression. FLG and TJ genes and proteins were assessed by quantitative RT-PCR, Western blot analysis, and immunofluorescent staining. The biotinylation reagent diffused through the paracellular spaces of whole stratified epithelial layers in EoE biopsy samples. The TEER decreased in ALI-cultured HEECs after IL-13 stimulation. Although the protein level of FLG decreased, that of the TJ proteins increased in the mucosa of EoE biopsy samples and in ALI-cultured HEECs after IL-13 stimulation. IL-13 altered the staining patterns of TJ proteins and the epithelial morphology. FLG siRNA transfection significantly decreased TEER. The IL-13-mediated reduced esophageal barrier is associated with the altered expression pattern but not with the levels of TJ-associated proteins. A deficiency of FLG altered the stratified epithelial barrier. NEW & NOTEWORTHY Esophageal permeability to small molecules was increased in patients with eosinophilic esophagitis (EoE) and could be induced by IL-13 in our unique air-liquid interface-cultured primary multilayer human esophageal epithelial cells in vitro. A deficiency of filaggrin disrupted the esophageal stratified epithelial barrier. The decreased esophageal barrier in EoE was associated with the altered staining pattern of tight junction proteins, although the levels of the proteins themselves do not appear to be changed.

scholarly journals Effects of metal nanoparticles on tight junction-associated proteins via HIF-1α/miR-29b/MMPs pathway in human epidermal keratinocytes

2021 ◽  
Vol 18 (1) ◽  
Author(s):  
Jiali Yuan ◽  
Yue Zhang ◽  
Yuanbao Zhang ◽  
Yiqun Mo ◽  
Qunwei Zhang
Keyword(s):  
Tight Junction ◽  
Metal Nanoparticles ◽  
Time Response ◽  
Nuclear Accumulation ◽  
Epidermal Keratinocytes ◽  
Hacat Cells ◽  
Down Regulation ◽  
Human Epidermal Keratinocytes ◽  
Associated Proteins ◽  
Dose Dependent

Abstract Background The increasing use of metal nanoparticles in industry and biomedicine raises the risk for unintentional exposure. The ability of metal nanoparticles to penetrate the skin ranges from stopping at the stratum corneum to passing below the dermis and entering the systemic circulation. Despite the potential health risks associated with skin exposure to metal nanoparticles, the mechanisms underlying the toxicity of metal nanoparticles on skin keratinocytes remain unclear. In this study, we proposed that exposure of human epidermal keratinocytes (HaCaT) to metal nanoparticles, such as nickel nanoparticles, dysregulates tight-junction associated proteins by interacting with the HIF-1α/miR-29b/MMPs axis. Methods We performed dose-response and time-response studies in HaCaT cells to observe the effects of Nano-Ni or Nano-TiO2 on the expression and activity of MMP-2 and MMP-9, and on the expression of tight junction-associated proteins, TIMP-1, TIMP-2, miR-29b, and HIF-1α. In the dose-response studies, cells were exposed to 0, 10, or 20 μg/mL of Nano-Ni or Nano-TiO2 for 24 h. In the time-response studies, cells were exposed to 20 μg/mL of Nano-Ni for 12, 24, 48, or 72 h. After treatment, cells were collected to either assess the expression of mRNAs and miR-29b by real-time PCR or to determine the expression of tight junction-associated proteins and HIF-1α nuclear accumulation by Western blot and/or immunofluorescent staining; the conditioned media were collected to evaluate the MMP-2 and MMP-9 activities by gelatin zymography assay. To further investigate the mechanisms underlying Nano-Ni-induced dysregulation of tight junction-associated proteins, we employed a HIF-1α inhibitor, CAY10585, to perturb HIF-1α accumulation in one experiment, and transfected a miR-29b-3p mimic into the HaCaT cells before Nano-Ni exposure in another experiment. Cells and conditioned media were collected, and the expression and activities of MMPs and the expression of tight junction-associated proteins were determined as described above. Results Exposure of HaCaT cells to Nano-Ni resulted in a dose-dependent increase in the expression of MMP-2, MMP-9, TIMP-1, and TIMP-2 and the activities of MMP-2 and MMP-9. However, exposure of cells to Nano-TiO2 did not cause these effects. Nano-Ni caused a dose-dependent decrease in the expression of miR-29b and tight junction-associated proteins, such as ZO-1, occludin, and claudin-1, while Nano-TiO2 did not. Nano-Ni also caused a dose-dependent increase in HIF-1α nuclear accumulation. The time-response studies showed that Nano-Ni caused significantly increased expressions of MMP-2 at 24 h, MMP-9 at 12, 24, and 48 h, TIMP-1 from 24 to 72 h, and TIMP-2 from 12 to 72 h post-exposure. The expression of miR-29b and tight junction-associated proteins such as ZO-1, occludin, and claudin-1 decreased as early as 12 h post-exposure, and their levels declined gradually over time. Pretreatment of cells with a HIF-1α inhibitor, CAY10585, abolished Nano-Ni-induced miR-29b down-regulation and MMP-2/9 up-regulation. Introduction of a miR-29b-3p mimic into HaCaT cells by transfection before Nano-Ni exposure ameliorated Nano-Ni-induced increased expression and activity of MMP-2 and MMP-9 and restored Nano-Ni-induced down-regulation of tight junction-associated proteins. Conclusion Our study herein demonstrated that exposure of human epidermal keratinocytes to Nano-Ni caused increased HIF-1α nuclear accumulation and increased transcription and activity of MMP-2 and MMP-9 and down-regulation of miR-29b and tight junction-associated proteins. Nano-Ni-induced miR-29b down-regulation was through Nano-Ni-induced HIF-1α nuclear accumulation. Restoration of miR-29b level by miR-29b-3p mimic transfection abolished Nano-Ni-induced MMP-2 and MMP-9 activation and down-regulation of tight junction-associated proteins. In summary, our results demonstrated that Nano-Ni-induced dysregulation of tight junction-associated proteins in skin keratinocytes was via HIF-1α/miR-29b/MMPs pathway.

scholarly journals The Human Cathelicidin LL-37 Host Defense Peptide Upregulates Tight Junction-Related Proteins and Increases Human Epidermal Keratinocyte Barrier Function

2014 ◽  
Vol 6 (6) ◽  
pp. 739-753 ◽  
Author(s):  
Toshihiro Akiyama ◽  
François Niyonsaba ◽  
Chanisa Kiatsurayanon ◽  
Toan The Nguyen ◽  
Hiroko Ushio ◽  
...  
Keyword(s):  
Tight Junction ◽  
Host Defense ◽  
Barrier Function ◽  
Epidermal Keratinocyte ◽  
Host Defense Peptide ◽  
Related Proteins ◽  
Human Epidermal Keratinocyte

scholarly journals MAPK pathway involved in epidermal terminal differentiation of normal human epidermal keratinocytes

Open Medicine ◽  
2018 ◽  
Vol 13 (1) ◽  
pp. 189-195 ◽  
Author(s):  
Xianguang Meng ◽  
Liyun Qiu ◽  
Haiyan Song ◽  
Ningning Dang
Keyword(s):  
Signaling Pathway ◽  
Mapk Pathway ◽  
Terminal Differentiation ◽  
Epidermal Keratinocytes ◽  
Human Epidermal Keratinocytes ◽  
Mapk Pathways ◽  
Normal Human Epidermal Keratinocytes ◽  
Normal Human ◽  
Related Proteins ◽  
Cytokeratin 14

AbstractObjectiveTo investigate the effect of mitogen-activated protein kinase (MAPK) signaling pathway in epidermal terminal differentiation.MethodsThe MAPK pathways (p38, ERK1/2, JNK) were inhibited by SB203580, PD98059, and SP600125 in normal human epidermal keratinocytes (NHEKs), respectively. Western blotting assays were performed to detect expression of filaggrin and differentiation-related proteins. The mRNA expressions of differentiation-related proteins were detected by real-time quantitative PCR (qRT-PCR).ResultsInhibition of MAPK pathway by SB203580, PD98059, and SP600125 resulted in significant reduction of filaggrin expression in NHEKs. Inhibition of the p38 MAPK pathway decreased the expression of differentiation-related proteins (cytokeratin 5, cytokeratin 14, ST14, and SPRR3), Akt, and NF-κB. Inhibition of JNK also suppressed expression of cytokeratin 14, SPRR3, Akt, and NF-κB. However, inhibition of ERK1/2 merely decreased expression of SPRR3 and Akt.ConclusionMAPK pathways regulates epidermal terminal differentiation in NHEKs. The p38 signaling pathway plays an especially important role.

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