Mechanistic studies on protective effects of total flavonoids from Ilex latifolia Thunb. on UVB-radiated human keratinocyte cell line (HaCaT cells) based on network pharmacology and molecular docking technique.

The aim of the present research is to investigate anti-UVB radiation activity of total flavonoids from Ilex latifolia Thunb. (namely large-leaved Kuding tea) on human keratinocyte cell line (HaCaT cells) based on network pharmacology and molecular docking technique. Network pharmacology was used to screen target genes of active ingredients from Ilex latifolia Thunb. associated with UVB irradiation. The possible signaling pathways were analyzed by KEGG enrichment and verified by cellular experiments. Molecular docking was used to assess the affinity between the active ingredients and the core targets. The prediction of network pharmacology and molecular docking was identified by series experiment in UVB-irradiated HaCaT cells. Network pharmacology results showed that the active ingredients of Ilex latifolia Thunb. for anti-UVB irradiation were mainly flavonoids, and the possible signaling pathways were involved in PI3K-AKT, apoptosis, MAPKs, NF-κB, and JAK-STAT3. Molecular docking indicated key binding activity between AKT1-Glycitein, STAT3-Formononetin, CASP3-Formononetin, TNF-Kaempferol, CASP3-Luteolin, and AKT1-Quercetin. The total flavonoid pretreatment (0.25-1.0 mg/mL) down-regulated the expression of IL-6, IL-1ß, and TNF-α in the cells determined by ELISA. The expression of phosphor PI3K, phosphor AKT, phosphor JAK, phosphor STAT3, phosphor JNK, and phosphor p38 MAPKs and COX-2 proteins in cytosolic and NF-κB p65 protein in nucleus were down-regulated and determined by western blot. It also protected UVB-irradiated cells from apoptosis by reducing apoptosis rate and down-regulating active-caspase 3. In a word, the total flavonoid treatment protected HaCaT cells from UVB injuries effectively, and the potential mechanism involves PI3K-AKT, JAK-STAT3, MAPK, and NF-κB pathway by anti-inflammatory and apoptosis action in cells. The mechanism in vivo experiment needs to be further confirmed in future.

Protective effect of isoquercitrin on UVB-induced injury in HaCaT cells and mice skin through anti-inflammatory, antioxidant, and regulation of MAPK and JAK2-STAT3 pathways.

Natural products are favored in the study of skin photodamage protection recently. Isoquercetin, namely 3-O-glucoside of quercetin, can be isolated from various plant species. In present research, the protective effect of isoquercitrin on UVB-induced injury in cells and mice skin were investigated. Our study reveals that 400 µM of isoquercitrin exhibits the best viability on UVB-irradiated HaCaT cells, and beneficial effects against oxidative stress UVB-induced in skin tissue by decreasing the levels of reactive oxygen species (ROS) and malondialdehyde (MDA), and simultaneously enhancing the activity of superoxide dismutase (SOD). Additionally, isoquercitrin was identified as an anti-inflammatory agent by reducing the level of COX-2 by Western blot analysis, and inflammatory cytokines such as IL-6, IL-1ß, and TNF-α by ELISA, and UVB-induced epidermal thickening evidenced by H&E staining. It also effectively prevented UVB-induced collagen fibers from degradation identified by Masson staining. Isoquercitrin significantly inhibited MAPK pathway by downregulating the levels of AP-1, MMP-1, MMP-3, phospho-p38, phospho-JNK, phospho-ERK, cleaved caspase-9, cleaved caspase-3, and JAK2-STAT3 pathway by western blot analysis. In conclusion, isoquercitrin pretreatment protected mice skin from UVB irradiation-induced injury effectively, and the underlying mechanism may involve MAPK and JAK2-STAT3 signaling pathways.

Natural products and extracts from plants as natural UV filters for sunscreens: A review.

Although solar exposure is necessary for human health, phototoxicology induced by excessive UVB and UVA radiation, which involves sunburns, skin aging and even tumorigenesis, has been widely researched. Sunscreen is one of the most important ways to protect skin from UV phototoxic damage. As well as inorganic and organic UV filters, some natural products or plant extracts with aromatic rings in their structures, such as flavonoids or polyphenols, can absorb UV to reduce sunburn, acting as a natural UV filter; they also show antioxidant or/and anti-inflammatory activity. This could explain why, although there are no officially approval natural commercial sun-filters, more and more commercial sunscreen products containing plant extracts are available on the market. Here we summarize articles focusing on natural UV filters from plant published in the last 6 years, selecting the most significant data in order to better understand the photoprotective activity of natural products and extracts from plants, including their major constituents and main biological effects, methods for evaluating UV radiation resistance, anti-UV radiation experimental models and anti-UV radiation mechanisms.

Bioaugmentation with GFP-Tagged Pseudomonas migulae AN-1 in Aniline-Contaminated Aquifer Microcosms: Cellular Responses, Survival and Effect on Indigenous Bacterial Community.

The recently isolated aniline-degrading bacterium Pseudomonas migulae AN-1 was tagged with green fluorescent protein (GFP) to investigate its bioaugmentation potential against anilinecontaminated groundwater through microcosm experiments. The survival and cellular response of GFP-tagged AN-1 introduced in a lab-scale aquifer corresponded directly with aniline consumption. During the process, the GFP-tagged AN-1 biomass increased from 7.52 × 105 cells/ml to 128 × 105 cells/ml and the degradation rate of aniline was 6.04 mg/l/h. GFP-tagged AN-1 was moderately hydrophobic (41.74%-47.69%) when treated with 20- 100 mg/l aniline and exhibited relatively strong hydrophobicity (55.25%-65.78%) when the concentration of aniline was ≥100 mg/l. The membrane permeability of AN-1 increased followed by a rise in aniline below 100 mg/l and was invariable with aniline above 100 mg/l. Pyrosequencing analysis showed that the relative abundance of Proteobacteria (accounted for 99.22% in the non-bioaugmentation samples) changed to 89.23% after bioaugmentation with GFP-tagged AN-1. Actinobacteria increased from 0.29% to 2.01%, whereas the abundance of Firmicutes barely changed. These combined findings demonstrate the feasibility of removing aniline in aquifers by introducing the strain AN-1 and provide valuable information on the changes in the diversity of dominant populations during bioaugmentation.