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

Abstract

Intrinsic skin aging and photoaging, from exposure to ultraviolet (UV) radiation, are associated with altered regulation of genes associated with the extracellular matrix (ECM) and inflammation, as well as cellular damage from oxidative stress. The regulatory properties of 1α, 25dihydroxyvitamin D3 (vitamin D) include endocrine, ECM regulation, cell differentiation, photoprotection, and anti-inflammation. The goal of this research was to identify the beneficial effects of vitamin D in preventing intrinsic skin aging and photoaging, through its direct effects as well as its effects on the ECM, associated heat shock proteins , cellular oxidative stress effects, and inflammatory cytokines [interleukin (IL)-1 and IL-8] in non-irradiated, UVA-radiated, UVB-radiated dermal fibroblasts. With regard to the ECM, vitamin D stimulated type I collagen and inhibited cellular elastase activity in non-irradiated fibroblasts; and stimulated type I collagen and HSP-47, and inhibited elastin expression and elastase activity in UVA-radiated dermal fibroblasts. With regard to cellular protection, vitamin D inhibited oxidative damage to DNA, RNA, and lipids in non-irradiated, UVA-radiated and UVB-radiated fibroblasts, and, in addition, increased cell viability of UVB-radiated cells. With regard to anti-inflammation, vitamin D inhibited expression of Il-1 and IL-8 in UVA-radiated fibroblasts, and stimulated HSP-70 in UVA-radiated and UVB-radiated fibroblasts. Overall, vitamin D is predominantly beneficial in preventing UVA-radiation induced photoaging through the differential regulation of the ECM, HSPs, and inflammatory cytokines, and protective effects on the cellular biomolecules. It is also beneficial in preventing UVB-radiation associated photoaging through the stimulation of cell viability and HSP-70, and the inhibition of cellular oxidative damage, and in preventing intrinsic aging through the stimulation of type I collagen and inhibition of cellular oxidative damage.Cosmetics 2019, 6, 46 2 of 15 and is degraded by elastases [1,11,[13][14][15][16][19][20][21][22][23]. Skin aging is associated with general atrophy of the ECM and reduced levels of collagen and elastin proteins and manifests as wrinkles and loss of skin firmness [1,3,9,[11][12][13][14][15][16]18,19]. Environmental factors, predominantly ultraviolet radiation (UV) radiation, superimpose on intrinsic skin aging to cause the loss of collagen fibers and the addition of elastotic deposits, which manifests as added wrinkles and coarse skin [1,3,12,14,16,[21][22][23]. The expression of collagen is inhibited by UVA-radiation (320-400 nm) and UVB-radiation (290-320 nm) in dermal fibroblasts [3,12]. The expression of elastin is stimulated by UVA radiation and inhibited by UVB radiation in dermal fibroblasts [1,14,16].The mechanisms of skin aging and photoaging includes oxidative stress from reactive oxygen species (ROS) and inflammation from inflammatory cytokines [4,5,7,8,[21][22][23][24][25][26][27][28][29][30]. The ROS are primarily superoxide, hydroxyl radicals, hydroge...

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].…”

“…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]. Vitamin D had been reported to inhibit oxidative stress, inflammation, angiogenesis and ECM remodeling in diverse cell types or tissue [30][31][32][33][34][35][36][37][38][39][40][41][42][43][44][45].…”

“…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 oxidative DNA/RNA damage was measured with the competitive DNA/RNA Oxidative Damage ELISA Kit (Cayman Chemical) [29]. Aliquots of cells or buffer were incubated with competitive acetylcholinesterase linked to 8-OH-dG (tracer), and antibody to oxidatively damaged guanine for 24 h at 4 • C, washed, incubated with substrate, and the product measured spectrophotometrically at 412 nm.…”

“…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.…”