Antioxidative and Anti-Aging Effects of Pinus rigida Mill. Ethyl Acetate Extract on the Human Dermal Fibroblast Cell Line CCD-986sk Damaged by Ultraviolet B Radiation
Journal Title: Biomedical Journal of Scientific & Technical Research (BJSTR) - Year 2019, Vol 12, Issue 4
Abstract
We analyzed the antioxidant and anti-wrinkle effects of Pinus rigida Mill. (pitch pine) to evaluate its use as a cosmetic material. 3-[4,5-Dimethylthiazol-2-yl]-2,5-diphenyl-tetrazoliumbromide assay was performed to evaluate the toxicity of the sample. To evaluate the anti-wrinkle activity of P. rigida Mill., the amount of type I procollagen and TIMP-1 synthesized was measured, and Western blot analysis and RT-PCR were performed to determine the expression levels of wrinkle-related proteins and genes. To investigate the inhibitory effect of ROS of P. rigida Mill, we conducted experiments to induce oxidative damage by UVB radiation using zebrafish embryos. Cytotoxicity to EtOAc extract (PRE) was measured using the human dermal fibroblast cell line CCD-986sk, and cytotoxicity was not observed below the concentration of 20 μg/mL. PRE has been shown to increase type I procollagen synthesis and promote the expression of its regulator, TIMP-1. In addition, P. rigida Mill. reduced the expression of MMP-1, -2, and -3 in CCD-986sk cells stimulated with UVB among the matrix metalloproteinases found in the dermal layers, which cause collagen reduction. After treatment with P. rigida Mill. extract at various concentrations, fluorescence spectra of ROS levels in the zebrafish embryos induced by UVB showed that fluorescence intensity decreased in a concentration-dependent manner compared with the intensity of the positive control group. Based on these findings, it is considered that P. rigida Mill. has value as a cosmetic material for antioxidant and anti-wrinkle benefits.Based on the characteristics of aging, it can be divided into endogenous and exogenous aging. Endogenous aging is caused by cell and tissue damage over time, abnormal immune function, and imbalance of epidermal homeostasis, whereas exogenous aging is caused by ultraviolet (UV) radiation, free radicals, toxicity, allergens, and physical stimuli [1]. A typical risk factor for exogenous aging is UVB, and constant exposure of the skin to it triggers fibroblasts to produce reactive oxygen species (ROS), including superoxide anion, peroxides, and singlet oxygen, which are considered harmful to humans. This reaction causes structural changes in the extracellular matrix, such as changes in collagen, elastin, proteoglycan, and fibronectin [2,3]. Excessively produced ROS can trigger DNA denaturation, which causes mutations in elastic fiber proteins and reduces collagen, leading to wrinkle formation and skin laxity [4]. Collagen reduction is caused by matrix metalloproteinase (MMP) in the dermal layers, which is activated by UV light [5,6]. MMPs, such as zinc-dependent peptides involved in the reconstruction of the extracellular matrix (endopeptidases), are related to skin cell death, skin ulcers, tumor invasion, and metastasis [7].
Authors and Affiliations
Young Ah Jang, Bo Ae Kim, Jin Tae Lee
Keywords
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- EP ID EP587451
- DOI 10.26717/BJSTR.2019.12.002285
- Views 147
- Downloads 0
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