Phlorizin, an Active Ingredient ofEleutherococcus senticosus, Increases Proliferative Potential of Keratinocytes with Inhibition of MiR135b and Increased Expression of Type IV Collagen

E. senticosusextract (ESE), known as antioxidant, has diverse pharmacologic effects. It is also used as an antiaging agent for the skin and phlorizin (PZ) is identified as a main ingredient. In this study, the effects of PZ on epidermal stem cells were investigated. Cultured normal human keratinocytes and skin equivalents are used to test whether PZ affects proliferative potential of keratinocytes and how it regulates these effects. Skin equivalents (SEs) were treated with ESE and the results showed that the epidermis became slightly thickened on addition of 0.002% ESE. The staining intensity of p63 as well as proliferating cell nuclear antigen (PCNA) is increased, and integrinα6 was upregulated. Analysis of ESE confirmed that PZ is the main ingredient. When SEs were treated with PZ, similar findings were observed. In particular, the expression of integrinα6, integrinβ1, and type IV collagen was increased. Levels of mRNA for type IV collagen were increased and levels of miR135b were downregulated. All these findings suggested that PZ can affect the proliferative potential of epidermal cells in part by microenvironment changes via miR135b downregulation and following increased expression of type IV collagen.

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Suppression of miR135b Increases the Proliferative Potential of Normal Human Keratinocytes

Journal of Investigative Dermatology ◽

10.1038/jid.2013.427 ◽

2014 ◽

Vol 134 (4) ◽

pp. 1161-1164 ◽

Cited By ~ 11

Author(s):

Hye-Ryung Choi ◽

Kyung-Mi Nam ◽

Sung-Jun Park ◽

Dong-Seok Kim ◽

Chang-Hun Huh ◽

...

Keyword(s):

Human Keratinocytes ◽

Proliferative Potential ◽

Normal Human Keratinocytes ◽

Normal Human

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The use of 1nm silver enhanced gold probes for the detection of skin antigens

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100162077 ◽

1990 ◽

Vol 48 (3) ◽

pp. 902-903

Author(s):

J.P Cassella ◽

H. Shimizu ◽

A. Ishida-Yamamoto ◽

R.A.J. Eady

Keyword(s):

Type Iv Collagen ◽

Freeze Substitution ◽

Collagen Type Iv ◽

Electron Microscopic ◽

Normal Human Skin ◽

Type Iv ◽

Type Vii Collagen ◽

Keratin Filaments ◽

Normal Human ◽

Phosphate Buffered Saline

1nm colloidal gold with silver enhancement has been used in conjunction with a low-temperature post-embedding (post-E) technique for the demonstration of skin antigens at both the light microscopic (LM) and electron microscopic (EM) levels.Keratin filaments and basement membrane zone (BMZ) associated antigens in normal human skin (NHS) were immunolabelled using antibodies against keratin 14, 10, and 1, the carboxy-terminus and collagenous portion of type VII collagen, type IV collagen and bullous pemphigoid antigen (BP-Ag).Fresh samples of NHS were cryoprotected in 15% glycerol, cryofixed in propane at -190°C, subjected to freeze substitution in methanol at -80°C and embedded in Lowicryl K11M at -60°C. Polymerisation of the resin was initiated under UVR at - 60°C for 48 hours and continued at room temperature for a further 48 hours. Semith in sections were air dried onto slides coated with 3-aminopropyltriethoxysilane. The following immunolabelling protocol was adopted: Primary antibody was applied for 2 hours at 37°C or overnight at 4°C. Following washing in Dulbecco’s phosphate buffered saline (PBSA) a biotinylated secondary antibody was applied for 2 hours at 37°C. The sections were further washed in PBSA and 1nm gold avidin was applied. Sections were finally washed in PBSA and silver enhanced.

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Differential expression of heparin-binding EGF-like growth factor (HB-EGF) mRNA in normal human keratinocytes induced by a variety of natural and synthetic retinoids

Experimental Dermatology ◽

10.1034/j.1600-0625.12.s2.5.x ◽

2003 ◽

Vol 12 (s2) ◽

pp. 28-34 ◽

Cited By ~ 23

Author(s):

Kotaro Yoshimura ◽

Gentaro Uchida ◽

Mutsumi Okazaki ◽

Yukie Kitano ◽

Kiyonori Harii

Keyword(s):

Growth Factor ◽

Differential Expression ◽

Human Keratinocytes ◽

Heparin Binding ◽

Normal Human Keratinocytes ◽

Normal Human ◽

Natural And Synthetic

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Hydrogen-Generating Silica Material Prevents UVA-Ray-Induced Cellular Oxidative Stress, Cell Death, Collagen Loss and Melanogenesis in Human Cells and 3D Skin Equivalents

Antioxidants ◽

10.3390/antiox10010076 ◽

2021 ◽

Vol 10 (1) ◽

pp. 76

Author(s):

Li Xiao ◽

Mai Mochizuki ◽

Taka Nakahara ◽

Nobuhiko Miwa

Keyword(s):

Oxidative Stress ◽

Cell Death ◽

Annexin V ◽

Human Keratinocytes ◽

Absorption Capacity ◽

Human Cells ◽

Skin Equivalents ◽

Type Iv ◽

Silica Material ◽

3D Skin

Ultraviolet-A (UVA) irradiation induces harmful effects on skin cells and accelerates skin aging through oxidative stress. In this study, the effects of a hydrogen-generating silica material named ULH-002 against UVA injuries in human cells and 3D skin equivalents were investigated. The oxygen radical absorption capacity (ORAC) assay showed that both freshly prepared ULH-002 solutions and 7-day-old solutions exhibited equal peroxyl radical (ROO·) scavenging activities concentration-dependently. CellROX® green/orange staining showed that ULH-002 could reduce UVA-induced oxidative stress in human keratinocytes HaCaT and human gingival fibroblasts (HGFs). ULH-002 significantly prevented UVA-induced apoptotic/necrotic cell death and cell-viability decline in HGFs and keratinocytes, as shown by Annexin V/PI apoptosis assay and PrestoBlue assay, respectively. Immunostaining showed that ULH-002 prevented the UVA-induced deterioration of expression of both type IV and I collagens in the 3D skin equivalents, and similarly in monolayer HGFs. UVA-enhanced melanogenesis was observed in human melanocytes HMV-II and HMV-II cell-containing 3D skin equivalents, but markedly prevented by ULH-002 as demonstrated by Fontana–Masson’s staining. In conclusion, our data suggested that ULH-002 could protect human keratinocytes and fibroblasts from UVA-induced injuries, prevent the loss of type IV and I collagens, as well as reduce melanogenesis. ULH-002 might be developed as a skin care reagent in the cosmetic industry.

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Nerve growth factor binds to normal human keratinocytes through high and low affinity receptors and stimulates their growth by a novel autocrine loop.

Journal of Biological Chemistry ◽

10.1016/s0021-9258(18)41604-3 ◽

1993 ◽

Vol 268 (30) ◽

pp. 22838-22846 ◽

Cited By ~ 1

Author(s):

E Di Marco ◽

M Mathor ◽

S Bondanza ◽

N Cutuli ◽

P.C. Marchisio ◽

...

Keyword(s):

Nerve Growth Factor ◽

Growth Factor ◽

Human Keratinocytes ◽

Nerve Growth ◽

Autocrine Loop ◽

Normal Human Keratinocytes ◽

Normal Human

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Effect of pro-inflammatory cytokines on the toxicity of the arylhydroxylamine metabolites of sulphamethoxazole and dapsone in normal human keratinocytes

Toxicology ◽

10.1016/j.tox.2005.10.002 ◽

2006 ◽

Vol 218 (2-3) ◽

pp. 90-99 ◽

Cited By ~ 15

Author(s):

F KHAN ◽

S ROYCHOWDHURY ◽

R NEMES ◽

P VYAS ◽

P WOSTER ◽

...

Keyword(s):

Inflammatory Cytokines ◽

Human Keratinocytes ◽

Normal Human Keratinocytes ◽

Normal Human ◽

Pro Inflammatory Cytokines

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UVB-mediated activation of p38 mitogen-activated protein kinase enhances resistance of normal human keratinocytes to apoptosis by stabilizing cytoplasmic p53

Biochemical Journal ◽

10.1042/bj20020072 ◽

2002 ◽

Vol 365 (1) ◽

pp. 133-145 ◽

Cited By ~ 96

Author(s):

Nadine CHOUINARD ◽

Kristoffer VALERIE ◽

Mahmoud ROUABHIA ◽

Jacques HUOT

Keyword(s):

Adaptive Response ◽

Human Keratinocytes ◽

Dominant Negative ◽

Mitogen Activated Protein Kinase ◽

Dominant Negative Mutant ◽

Normal Human Keratinocytes ◽

Mitogen Activated Protein ◽

Normal Human ◽

Induced Apoptosis ◽

Negative Mutant

Human keratinocytes respond to UV rays by developing a fast adaptive response that contributes to maintaining their functions and survival. We investigated the role of the mitogen-activated protein kinase pathways in transducing the UV signals in normal human keratinocytes. We found that UVA, UVB or UVC induced a marked and persistent activation of p38, whereas c-Jun N-terminal kinase or extracellular signal-regulated kinase were less or not activated respectively. Inhibition of p38 activity by expression of a dominant-negative mutant of p38 or with SB203580 impaired cell viability and led to an increase in UVB-induced apoptosis. This sensitization to apoptosis was independent of caspase activities. Inhibition of p38 did not sensitize transformed HaCaT keratinocytes to UVB-induced apoptosis. In normal keratinocytes, expression of a dominant-negative mutant of p53 increased UVB-induced cell death, pointing to a role for p53. In these cells, UVB triggered a p38-dependent phosphorylation of p53 on Ser-15. This phosphorylation was associated with an SB203580-sensitive accumulation of p53, even in the presence of a serine phosphatase inhibitor. Accumulated p53 was localized mainly in the cytoplasm, independently of CRM1 nuclear export. In HaCaT cells, p53 was localized exclusively in the nucleus and its distribution and level were not affected by UVB or p38 inhibition. However, UVB induced an SB203580-insensitive phosphorylation on Ser-15 of mutated p53. Overall, our results suggest that, in normal human keratinocytes, protection against UVB depends on p38-mediated phosphorylation and stabilization of p53 and is tightly associated with the cytoplasmic sequestration of wild-type p53. We conclude that the p38/p53 pathway plays a key role in the adaptive response of normal human keratinocytes against UV stress.

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