Enzyme-Treated Asparagus Extract Attenuates Hydrogen Peroxide-Induced Matrix Metalloproteinase-9 Expression in Murine Skin Fibroblast L929 Cells

Abstract

Enzyme-treated asparagus extract (ETAS) exerts a wide variety of beneficial biological actions including facilitating anti-cortisol stress and neurological antiaging responses. However, the anti-skin aging effects of ETAS remain to be elucidated. Reactive oxygen species (ROS) play pivotal roles in skin aging. Increased ROS levels in fibroblasts in response to ultraviolet irradiation activate c-Jun N-terminal kinase (JNK) and its downstream transcription factor activator protein-1 (AP-1), and the resultant gene expression of matrix metalloproteinase (MMP) isoforms accelerates collagen breakdown in the dermis. Therefore, we explored whether ETAS has anti-skin aging effects by attenuating the oxidative stress responses in fibroblasts. Simultaneous treatment of murine skin L929 fibroblasts with hydrogen peroxide (H 2 O 2) and either ETAS or dextrin showed that ETAS significantly suppressed H 2 O 2-induced expression of MMP-9 mRNA as measured by real-time polymerase chain reaction. ETAS also clearly suppressed H 2 O 2-stimulated phosphorylation of c-Jun (AP-1 subunit) and JNK as determined by Western blot. However, ETAS did not affect the increased amounts of carbonyl proteins in response to H 2 O 2 , also as determined by Western blotting. These results suggest that ETAS diminishes cellular responsiveness to ROS but does not scavenge ROS. Thus, ETAS has the potential to prevent skin aging through attenuating the oxidative stress responses in dermal fibroblasts.

Skin Anti-aging Assays of Proanthocyanidin Rich Red Rice Extract, Oryzanol and Other Phenolic Compounds

“…The main enzymes involved in extracellular matrix (ECM) degradation are elastase and matrix metalloproteinases (MMPs). Specific examples of these are gelatinases (MMP-2) and collagenase [2][3]. MMPs activity is accelerated with age and in the presence of reactive oxygen species [2], particularly superoxide anion which is a potent inflammatory mediator [4].…”

Enzyme-Treated Asparagus Extract Prevents Hydrogen Peroxide-Induced Pro-Inflammatory Responses by Suppressing p65 Nuclear Translocation in Skin L929 Fibroblasts

“…Cell culture: The murine skin fibroblast cell line L929 (DS Pharma Biomedical, Osaka, Japan) was cultured as previously described [20]. Twenty-four h prior to the initiation of experiments, cells were seeded on culture dishes or multi-well plates at a density of 3.0 × 10 4 cells/cm 2 [20].…”

“…Cell culture: The murine skin fibroblast cell line L929 (DS Pharma Biomedical, Osaka, Japan) was cultured as previously described [20]. Twenty-four h prior to the initiation of experiments, cells were seeded on culture dishes or multi-well plates at a density of 3.0 × 10 4 cells/cm 2 [20]. To test the effect of H 2 O 2 alone, the cells were treated with or without 1 mM H 2 O 2 (Sigma-Aldrich, St. Louis, MO, USA) and cultured for the desired periods [20].…”

“…Twenty-four h prior to the initiation of experiments, cells were seeded on culture dishes or multi-well plates at a density of 3.0 × 10 4 cells/cm 2 [20]. To test the effect of H 2 O 2 alone, the cells were treated with or without 1 mM H 2 O 2 (Sigma-Aldrich, St. Louis, MO, USA) and cultured for the desired periods [20]. To assess the effect of ETAS, either ETAS or dextrin (vehicle) were directly dissolved in complete medium to produce a final concentration of 1 mg/mL, and then the supplemented medium was sterilized using a Millex 0.22-μm Syringe-Driven Filter Unit (Merck Millipore, Darmstadt, Germany).…”

Enzyme-treated Asparagus Extract (ETAS) Enhances Memory in Normal Rats and Induces Neurite-outgrowth in PC12 Cells

“…Enzyme-treated Asparagus Extract (ETAS) is a manufacturing product made from the residual lower parts of asparagus [2]. Some studies have reported about functionalities of ETAS such as anti-inflammatory [3], antioxidative activities [4], the protective effects on early cognitive impairment of senescence-accelerated mouse prone 8 (SAMP8) mice and β-amyloid (Aβ)-induced reduction of PC12 cell (rat pheochromocytoma) viability [5,6].…”