Here, we determined the immunostimulatory effects of black radish (Raphanus sativus ver niger) hot water extract (BRHE) on a mouse macrophage cell line (RAW 264.7) and mouse peritoneal macrophages. We found that BRHE treatment increased cell proliferation, phagocytic activity, nitric oxide (NO) levels, cytokine production, and reactive oxygen species synthesis. Moreover, BRHE increased the expression of the following immunomodulators in RAW 264.7 cells and peritoneal macrophages: pro-inflammatory cytokines (IL-1β, IL-6, and TNF-α), iNOS, and COX-2. BRHE treatment significantly up-regulated the phosphorylation of components of the mitogen-activated protein kinase (MAPK), nuclear factor-κB (NF-κB), Akt, and STAT3 signaling pathways. Further, the effects of BRHE on macrophages were significantly diminished after the cells were treated with the TLR2 antagonist C29 or the TLR4 antagonist TAK-242. Therefore, BRHE-induced immunostimulatory phenotypes in mouse macrophages were reversed by multiple inhibitors, such as TLR antagonist, MAPK inhibitor, and Akt inhibitor indicating that BRHE induced macrophage activation through the TLR2/4–MAPK–NFκB–Akt–STAT3 signaling pathway. These results indicate that BRHE may serve as a potential immunomodulatory factor or functional food and provide the scientific basis for the comprehensive utilization and evaluation of black radish in future applications.
Schizophragma hydrangeoides (S. hydrangeoides) is a vine endogenous to Jeju Island and Ulleungdo, where it grows attached to the foothills and rock surfaces. Previous research has mostly focused on the whitening effect of S. hydrangeoides leaf extract. In this study, we investigated S. hydrangeoides leaf extract further, and detected four phytochemicals in the extract: chlorogenic acid, quercetin-3-O-glucosyl-(1-2)-rhamnoside, quercetin-3-Oxylosyl-(1-2)-rhamnoside, and quercitrin. We pretreated human dermal fibroblast (HDFn) cells with previously established concentrations of the four compounds for 1 h before ultraviolet A (UVA) irradiation. Among the four compounds, quercetin-3-Oglucosyl-(1-2)-rhamnoside (Q-3-GR) best inhibited matrix metalloproteinase-1 (MMP-1) levels. Thus, we investigated the protective effects of Q-3-GR on photoaging and its underlying mechanisms. Q-3-GR significantly reduced MMP-1 production and inhibited MMP-1 protein expression in UVA-irradiated HDFn cells. Furthermore, Q-3-GR increased procollagen type I production and protein expression. Q-3-GR exerted its anti-photoaging effects by downregulating the mitogen-activated protein kinase/ activator protein-1 signaling pathway, and upregulating the transforming growth factor-β/Smad signaling pathway. Thus, S. hydrangeoides leaf-derived Q-3-GR is a potential potent cosmetic ingredient for UV-induced skin aging.
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