Beneficial Regulation of Elastase Activity and Expression of Tissue Inhibitors of Matrixmetalloproteinases, Fibrillin, Transforming Growth Factor-β, and Heat Shock Proteins by P. leucotomos in Nonirradiated or Ultraviolet-Radiated Epidermal Keratinocytes

Abstract

There is loss of the structural integrity of the extracellular matrix (ECM) with intrinsic aging as well as photoaging, largely due to reactive oxygen species (ROS). The structural ECM proteins include the microfibrils that are composed of fibrillin. The structural ECM proteins are primarily degraded by the matrixmetalloproteinases (MMPs) and elastase enzymes. The MMPs are inhibited by the tissue inhibitors of MMPs (TIMPs). A primary regulator of the ECM proteins is transforming growth factor-(TGF-), and the chaperone proteins important for its formation are the heat shock proteins (HSP). P. leucotomos extract beneficially regulates of MMPs, TIMPs, and TGF-in nonirradiated or ultraviolet (UV) radiated fibroblasts and melanoma cells. The hypothesis of this research was that the antioxidant activity or chemistry of P. leucotomos extract would also directly inhibit elastase activity, stimulate the cellular expression of TIMPs, fibrillins, and TGF-, and regulate HSPs in nonirradiated and UVA or UVB radiated epidermal keratinocytes. P. leucotomos directly inhibited elastase activity, stimulated the cellular expression of TIMPs, fibrillins, and TGF-, and differentially regulated HSPs in nonirradiated and UVA or UVB radiated epidermal keratinocytes. We infer that the P. leucotomos extract strengthens the ECM and is effective in the prevention or treatment of intrinsic and photoaging of skin.

Beneficial Regulation of Cellular Oxidative Stress Effects, and Expression of Inflammatory, Angiogenic, and the Extracellular Matrix Remodeling Proteins by 1α,25-Dihydroxyvitamin D3 in a Melanoma Cell Line

“…The alterations collectively enable the cancer cells to grow and metastasize. We have reported the beneficial regulation of cellular and extracellular parameters by plant extracts, phenolic compounds, hormones, and vitamins in the amelioration of aging, photoaging or photocarcinogenesis [21][22][23][24][25][26][27][28][29][46][47][48][49][50][51][52]. We recently reported the inhibition of cellular oxidative stress effects and inflammation by vitamin D in non-irradiated, and UV radiated fibroblasts [29].…”

“…The cells were examined for cell viability by the CellTiter 96 ® Aqueous One Solution reagent (tetrazolium compound (3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium, inner salt; MTS) and electron coupling reagent (phenazine ethosulfate; PES)] (Promega); by incubating the cells, after the 24 h of incubation with vitamin D, with aliquots of the MTS reagent (yellow) for 30 min at 37 • C, and measuring the product (brown, through the conversion of the tertrazolium reagent to formazan by viable cells) spectrophotometrically at 490 nm [21][22][23][24][25][26][27][28][29][46][47][48][49][50][51][52].…”

“…The expression of SOD, IL-1, TNF-α, TGF-β, VEGF, MMP-1, and MMP-2 proteins was measured by indirect ELISA (Kirkguaard and Perry Laboratories, Inc) [21][22][23][24][25][26][27][28][29][46][47][48][49][50][51][52]. The aliquots of cells or media were incubated with coating buffer in 96-well immunosorbent plates for 24 h at 4 • C, blocked with bovine serum albumin for 30 min at room temperature, incubated with respective primary antibodies (Cayman Chemical, Millipore, Sigma, St.Louis, MO, USA) for 1 h at room temperature, washed with wash buffer, incubated with secondary antibody linked to peroxidase for 1 h at room temperature, washed, and incubated with peroxidase substrate until color development, and measured spectrophotometrically at 405 nm.…”

The Beneficial Regulation of Extracellular Matrix and Heat Shock Proteins, and the Inhibition of Cellular Oxidative Stress Effects and Inflammatory Cytokines by 1α, 25 dihydroxyvitaminD3 in Non-Irradiated and Ultraviolet Radiated Dermal Fibroblasts

“…The direct inhibitory effect of vitamin D on elastase activity was determined by an elastase activity inhibition assay, which is based on the inhibition of the conversion of an elastase substrate to a colored product (Elastin products Co, Owensville, Missouri) [1,6,11]. Vitamin D at 0, 0.02, 0.2, or 2 µM was incubated with elastase for 10 min followed by the addition of its substrate, and the product was measured spectrophotometrically at 410 nm.…”

“…The structural integrity of the extracellular matrix (ECM) is essential to anti-aging [1][2][3][4][5][6][7][8][9][10][11][12][13][14][15][16]. The structural ECM is composed largely of collagen and elastin fibers, which are synthesized by the dermal fibroblasts [9][10][11][12][13][14][15][16].…”