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1.
J Microbiol Biotechnol ; 27(5): 933-938, 2017 May 28.
Article in English | MEDLINE | ID: mdl-28297750

ABSTRACT

Clitocybin A, an isoindolinone from Clitocybe aurantiaca, was investigated to assess its anti-wrinkle properties, through reactive oxygen species (ROS)-scavenging and elastase inhibitory activities, procollagen synthesis, and matrix metalloproteinase-1 (MMP-1) expression, in human primary dermal fibroblast-neonatal (HDF-N) cells. Clitocybin A exhibited no significant cytotoxicity up to 10 ppm in HDF-N cells, with cell viability and cell proliferation activity greater than 94.6% and 91.9%, respectively. Strong and concentration-dependent ROS radical scavenging activities of clitocybin A were observed following irradiation with UVB at 30 mJ/cm2. Furthermore, clitocybin A treatment of cells at 0.1, 1, and 10 ppm exhibited decreased elastase activity, in a concentration-dependent manner, by 1.97%, 6.6%, and 8.31%, respectively, versus the control group. The effects of clitocybin A on procollagen synthesis and MMP-1 expression were investigated. Clitocybin A treatment of cells at 1, 5, and 10 ppm increased procollagen synthesis, by 67.9%, 74.4%, and 112.9%, respectively, versus the control group. At these concentrations, MMP-1 expression decreased significantly following UV irradiation. Together, these findings suggest that clitocybin A may be an effective ingredient for use in anti-wrinkle cosmetic products.


Subject(s)
Agaricales/chemistry , Free Radical Scavengers/pharmacology , Isoindoles/antagonists & inhibitors , Mycelium/metabolism , Reactive Oxygen Species/metabolism , Cell Cycle/drug effects , Cell Line/drug effects , Cell Line/metabolism , Cell Line/radiation effects , Cell Proliferation/drug effects , Cell Survival/drug effects , Cells, Cultured , Fibroblasts/drug effects , Fibroblasts/radiation effects , Humans , Isoindoles/administration & dosage , Isoindoles/chemistry , Matrix Metalloproteinase 1/biosynthesis , Matrix Metalloproteinase 1/drug effects , Matrix Metalloproteinase 1/radiation effects , Pancreatic Elastase/drug effects , Pancreatic Elastase/metabolism , Procollagen/antagonists & inhibitors , Procollagen/biosynthesis , Procollagen/radiation effects , Reactive Oxygen Species/radiation effects , Scattering, Radiation , Skin Aging/drug effects , Skin Aging/radiation effects , Ultraviolet Rays
2.
Regul Toxicol Pharmacol ; 60(1): 112-9, 2011 Jun.
Article in English | MEDLINE | ID: mdl-21397653

ABSTRACT

UV exposure is known to induce premature aging, which is mediated by matrix metalloproteinase-1 (MMP-1) activity. MMP-1 mRNA expression is up-regulated by elevated cysteine-rich 61 (CYR61) and monocyte chemoattractant protein-1 (MCP-1) via action of transcription factor AP-1. Collagen is degraded by MMP-1 activity but synthesized by transforming growth factor-ß (TGF-ß) signal. Chlorella has been shown to inhibit UVB-induced MMP-1 level, however its regulatory molecular mechanisms have not been studied. In this study, Chlorella derived peptide (CDP) was added to skin fibroblasts after UVB irradiation and the expression of MMP-1, CYR61, procollagen, c-fos, c-jun, and TGF-ß receptor (TbRII) mRNA and MCP-1 production were investigated. CDP (10 or 5mg/ml) diminished UVB-induced MMP-1 and CYR61 mRNA expression and MCP-1 production, whereas, UVB-suppressed procollagen and TbRII mRNA was restored by CDP treatment. UVB-induced c-fos and c-jun expressions were also inhibited by the CDP treatment. Taken together, CDP inhibits UVB-induced MMP-1 expression in skin fibroblasts by suppressing expression of AP-1 and CYR61 and MCP-1 production.


Subject(s)
Chlorella/chemistry , Fibroblasts/drug effects , Gene Expression/drug effects , Matrix Metalloproteinase 1/metabolism , Peptides/pharmacology , Procollagen/metabolism , Skin/drug effects , Cell Proliferation/drug effects , Cell Proliferation/radiation effects , Cell Survival/drug effects , Cell Survival/radiation effects , Cells, Cultured , Chemokine CCL2/genetics , Chemokine CCL2/metabolism , Cysteine-Rich Protein 61/genetics , Cysteine-Rich Protein 61/metabolism , Fibroblasts/metabolism , Fibroblasts/radiation effects , Gene Expression/radiation effects , Humans , Matrix Metalloproteinase 1/genetics , Peptides/isolation & purification , Plant Extracts/pharmacology , Plant Proteins/isolation & purification , Plant Proteins/pharmacology , Procollagen/genetics , Procollagen/radiation effects , Skin/metabolism , Skin/radiation effects , Transcription Factor AP-1/genetics , Transcription Factor AP-1/metabolism , Ultraviolet Rays
3.
Biomol Eng ; 24(5): 447-54, 2007 Nov.
Article in English | MEDLINE | ID: mdl-17825608

ABSTRACT

Adhesion and spreading of cells on biomaterials are integrin-mediated processes. But recent findings indicate a key role of the cell membrane associated matrix substance hyaluronan (HA) in interface interactions. Because HA is a negatively charged molecule we assume that a biomaterial surface with an opposed charge could boost the first contact of the cell to the surface. Polished cp titanium (R(a)=0.19 microm) was coated with an amino-group containing plasma polymer (Ti PPA). For this purpose, a microwave excited, pulsed, low-pressure plasma was used. Additionally, collagen was immobilized on Ti PPA with polyethylene glycol diacid (PEG-DA), catalyzed by carbodiimide (CDI). The physico-chemical surface analytical techniques like XPS, FT-IR, water contact angle and zeta-potential verified the retention of the allylamine precursor structure. Human osteoblasts were cultured in serum-free Dulbecco's modified Eagle medium (DMEM). Adhesion and cell cycle phases were calculated by flow cytometry. Spreading and actin cytoskeleton were visualized by confocal microscopy. Gene expression of osteogenic markers was detected by real-time RT-PCR. Ti PPA is significantly advantageous concerning initial adhesion and spreading during the first hours of the cell contact to the surface. The proliferation of osteoblasts is positively influenced. Gene expression of the differentiation marker bone sialoprotein was upregulated after 24h. Our results demonstrate that functionalization of titanium with positively charged amino-groups is sufficiently enough to significantly improve initial steps of the cellular contact to the material surface.


Subject(s)
Allylamine/chemistry , Osteoblasts/physiology , Polymers/chemistry , Titanium/chemistry , Actins/chemistry , Alkaline Phosphatase/genetics , Carbodiimides/chemistry , Catalysis , Cell Adhesion/physiology , Cell Cycle , Collagen Type I/chemistry , Collagen Type I/genetics , Collagen Type I/radiation effects , Cytoskeleton/chemistry , Flow Cytometry , Gene Expression Profiling , Humans , Microwaves , Polyethylene Glycols/chemistry , Polyethylene Glycols/radiation effects , Polymers/radiation effects , Procollagen/chemistry , Procollagen/genetics , Procollagen/radiation effects , Reverse Transcriptase Polymerase Chain Reaction , Surface Properties , Titanium/radiation effects , Tumor Cells, Cultured
4.
J Lipid Res ; 47(5): 921-30, 2006 May.
Article in English | MEDLINE | ID: mdl-16467281

ABSTRACT

Skin aging can be attributed to photoaging (extrinsic) and chronological (intrinsic) aging. Photoaging and intrinsic aging are induced by damage to human skin attributable to repeated exposure to ultraviolet (UV) irradiation and to the passage of time, respectively. In our previous report, eicosapentaenoic acid (EPA) was found to inhibit UV-induced matrix metalloproteinase-1 (MMP-1) expression in human dermal fibroblasts. Therefore, we investigated the effects of EPA on UV-induced skin damage and intrinsic aging by applying EPA topically to young and aged human skin, respectively. By immunohistochemical analysis and Western blotting, we found that topical application of EPA reduced UV-induced epidermal thickening and inhibited collagen decrease induced by UV light. It was also found that EPA attenuated UV-induced MMP-1 and MMP-9 expression by inhibiting UV-induced c-Jun phosphorylation, which is closely related to UV-induced activator protein-1 activation, and by inhibiting JNK and p38 activation. EPA also inhibited UV-induced cyclooxygenase-2 (COX-2) expression without altering COX-1 expression. Moreover, it was found that EPA increased collagen and elastic fibers (tropoelastin and fibrillin-1) expression by increasing transformin growth factor-beta expression in aged human skin. Together, these results demonstrate that topical EPA has potential as an anti-skin-aging agent.


Subject(s)
Eicosapentaenoic Acid/pharmacology , Radiation-Protective Agents/pharmacology , Skin Aging/drug effects , Adult , Aged , Cyclooxygenase 2/biosynthesis , Extracellular Matrix/drug effects , Gene Expression/radiation effects , Humans , MAP Kinase Kinase 4/metabolism , MAP Kinase Kinase 4/radiation effects , Male , Matrix Metalloproteinase 1/biosynthesis , Matrix Metalloproteinase 9/biosynthesis , Phosphorylation/radiation effects , Procollagen/biosynthesis , Procollagen/radiation effects , Proto-Oncogene Proteins c-jun/metabolism , Proto-Oncogene Proteins c-jun/radiation effects , Skin Aging/radiation effects , Transforming Growth Factor beta/biosynthesis , Transforming Growth Factor beta1 , Transforming Growth Factor beta2 , Transforming Growth Factor beta3 , Ultraviolet Rays , p38 Mitogen-Activated Protein Kinases/metabolism , p38 Mitogen-Activated Protein Kinases/radiation effects
5.
Int J Radiat Biol ; 75(11): 1429-36, 1999 Nov.
Article in English | MEDLINE | ID: mdl-10597916

ABSTRACT

PURPOSE: To study alteration in gene transcription (transforming growth factor-beta 1 and procollagen types I and III) involved in radiation-induced cardiac damage. MATERIALS AND METHODS: Female Sprague-Dawley rats were irradiated with a single dose of 0, 15, 20 or 25 Gy locally on the heart. At intervals up to 16 months after irradiation, absolute amounts of mRNA were quantified using a (semi-nested) competitive PCR assay. All values were normalized to equal input cDNA with respect to their GAPDH content. RESULTS: After irradiation, left ventricular TGF-beta 1 mRNA levels increased sharply. This response was bi-phasic with peaks at days 1 and 12 (maximum 6-fold baseline), then returning to control levels by 1 month. After 20 Gy, a persistent elevation was observed from 6 months, but this elevation was less profound (approximately 1.5-fold baseline) when compared with the early response (1-12 days). Absolute mRNA levels of procollagen type I hardly changed during the first 6 months, but thereafter these levels increased progressively until the end of observation. An age-related increase in procollagen I was also observed. Procollagen type III mRNA levels were increased between days 1 and 12, returned to control values and remained low up to 6 months, then mRNA levels rose again with increasing time post-treatment. CONCLUSION: The difference in time-course between TGF-beta 1 and procollagen mRNA expression after local heart irradiation and ageing strongly suggest that the late up-regulation of both procollagen types in the left ventricle occurs without TGF-beta 1 over-expression.


Subject(s)
Heart/radiation effects , Myocardium/metabolism , Procollagen/biosynthesis , RNA, Messenger/biosynthesis , RNA, Messenger/radiation effects , Transforming Growth Factor beta/biosynthesis , Animals , Cardiomyopathies/genetics , Dose-Response Relationship, Radiation , Female , Procollagen/genetics , Procollagen/radiation effects , RNA, Messenger/metabolism , Radiation Injuries, Experimental/genetics , Rats , Rats, Sprague-Dawley , Transcription, Genetic/radiation effects , Transforming Growth Factor beta/genetics , Transforming Growth Factor beta/radiation effects , Ventricular Function, Left/radiation effects
6.
Aust Dent J ; 41(3): 188-92, 1996 Jun.
Article in English | MEDLINE | ID: mdl-8768644

ABSTRACT

A low power Ga-As pulse laser was used to stimulate cultured human embryonic fibroblast cells. Energy fluencies varied from 0-1 J/cm2 over a period of 1-4 days. Fibroblast procollagen production was monitored by the synthesis of [3H] hydroxyproline, and DNA replication was assessed by [3H] thymidine incorporation. Following laser treatment, controlled pepsin digestion measured the increase in cell biostimulation. Maximum increase in collagen production and cell biostimulation occurred after 4 episodes of laser treatment at 24-hour intervals. Laser doses between 0.099 and 0.522 J/cm2 had the most significant stimulatory effects on fibroblast function. Clinical efficacy of the low power Ga-As pulse laser may be related to enhanced connective tissue repair.


Subject(s)
Collagen/metabolism , Fibroblasts/metabolism , Lasers , Arsenic , Cells, Cultured , Collagen/radiation effects , Connective Tissue/metabolism , Connective Tissue/radiation effects , DNA Replication/radiation effects , Embryo, Mammalian , Fibroblasts/radiation effects , Gallium , Humans , Hydroxyproline/biosynthesis , Hydroxyproline/radiation effects , Pepsin A , Procollagen/biosynthesis , Procollagen/metabolism , Procollagen/radiation effects , Thymidine/metabolism , Thymidine/radiation effects , Time Factors , Tritium
7.
Photodermatol Photoimmunol Photomed ; 10(5): 212-6, 1994 Oct.
Article in English | MEDLINE | ID: mdl-7880761

ABSTRACT

The effect of aging, sex, skin location and a short whole body ultraviolet B (UVB) phototherapy on type I and type III skin collagen synthesis were studied by measuring carboxy- and aminoterminal propeptides of type I and aminoterminal propeptide of type III procollagens (PICP, PINP and PIIINP, respectively) in suction blister fluid (SBF). The concentrations of PICP and PIIINP correlated negatively with age in the material of 30 men and 27 women (age range 23-86 years, mean age 51 years). As the subjects were divided into 3 age groups (23-34, 35-54 and 55-86 years) and the mean concentrations of the procollagens were compared between the groups, the youngest age groups in men and in the whole material had significantly higher concentrations than the two older age groups. These results confirm the previous results, which have shown by other methods that skin collagen synthesis decreases during aging. SBF was also obtained from 4 different body regions (back, arm, abdomen and leg) of 5 young male subjects and from 2 different body regions (abdomen and thigh) of 9 older subjects. Wide individual range in the concentrations of PICP, PINP and PIIINP was noticed but, in general, none of the body regions seemed to have more excessive collagen synthesis than the other ones. SBF was further collected from abdominal skin of 8 young male dermatological patients receiving UVB phototherapy. When the concentrations of PICP and PIIINP in SBF obtained before the treatment, after the treatment and from the covered control area were compared no statistical differences were noted, suggesting that short UVB treatment has no effect on skin collagen synthesis in vivo.


Subject(s)
Aging/metabolism , Collagen/biosynthesis , Skin/metabolism , Ultraviolet Rays , Abdomen , Adult , Aged , Aged, 80 and over , Aging/pathology , Arm , Back , Blister/metabolism , Collagen/radiation effects , Cross-Sectional Studies , Exudates and Transudates/metabolism , Female , Humans , Leg , Male , Middle Aged , Peptide Fragments/biosynthesis , Peptide Fragments/radiation effects , Procollagen/biosynthesis , Procollagen/radiation effects , Sex Factors , Skin/anatomy & histology , Skin/radiation effects , Ultraviolet Therapy
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