Abstract
Clinical studies have identified that an excessive exposure of UV light can cause oxidative stress (OS) and tyrosinase enzyme over-expression, which are associated with multiple diseases including atherosclerosis, cancers, diabetics, rheumatoid arthritis. In this study, we investigated the impact of grape seed proanthocyanidin (GSPE) on regulating OS and tyrosinase activity in human epidermal melanocytes. This study revealed that GSPE did not affect cell viability and protected cells from UV induced damage in a dose-dependent manner. 5-(-6)-Carboxy-2,7-di-chlorodihydro-fluorescein diacetate staining (i.e., a fluorescence staining for intracellular (OS)) indicated that GSPE reduced OS level caused by UV exposure. A similar trend was also confirmed by flow cytometry analysis, where GSPE down-regulated OS level. Tyrosinase analysis showed that GSPE treatment decreased tyrosinase activity. Taken all data together, GSPE may restore the cellular damage caused by excessive UV-exposure and promote skin health by reducing tyrosine generation. Clinically, GSPE could be potentially utilized for improving skin health against excessive UV exposure.
Highlights
Human skin is the first line of protection against toxicants or their metabolites, which are usually oxidants that are associated with the production of reactive oxidants or reactive oxygen species (ROS), reactive substances that are continuously generated at low levels during metabolism.[1]
The test illustrated that grape seed proanthocyanidin extract (GSPE) under different concentrations had similar viability: the cells treated with 3, 10, 25, and 50 μg mL-1 GSPE had 93.0 ± 2.1%, 89.1 ± 1.9%, 91.2 ± 3.5%, and 90.2 ± 2.0% viable cells, respectively, compared to 91.2 ± 3.5% live cells in the untreated samples (p > 0. 05 among all groups, Figure 1a). This data indicated that GSPE did not affect cell viability. This trend was confirmed by live/dead cell staining under fluorescence microscope: the cells were treated with 25 μg mL-1 GSPE for 48 h, and the cells were still alive as indicated in green color (Figure 1b); as a contrast, a very limited number of dead cells were observed under fluorescence microscope, as indicated in red color in Figure 1b, which happened naturally since cells will die in their cellular cycle
This work studied the impact of GSPE on cell viability, proliferation, UV-induced reactive oxygen stress and tryosinase production
Summary
Human skin is the first line of protection against toxicants or their metabolites, which are usually oxidants that are associated with the production of reactive oxidants or reactive oxygen species (ROS), reactive substances that are continuously generated at low levels during metabolism.[1]. Studies have reported that the production of these oxidants can induce acute responses to the immune cells including macrophages, neutrophils, enhancing the expression of vascular endothelial growth factor that can potentially assist wound healing in the skin.[2] In clinic, the increased level of intracellular oxidative stress within human bodies is associated with multiple types of diseases including atherosclerosis, cancer, diabetics, rheumatoid. Multiple studies[8,9] reported using grape seed proanthocyanidin extract (GSPE) to inhibit oxidative stress for different purposes including cancer treatment, immune protection and skin pigment removal. We focused on studying the impact of GSPE to the intracellular levels of ROS and tyrosinase activity. We employed GSPE to regulate the production of ROS and tyrosinase enzyme induced by UV exposure. This study illustrated that GSPE can down-regulate ROS and tyrosinase activity in the UV-exposure environment, indicating that GSPE could improve skin health in clinic. Dimethyl sulfoxide (DMSO) was from the Sigma Chemical (Shanghai, China)
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