In vitro antioxidant and in vivo anti-inflammatory activities of Ophioglossum thermale.

Antioxidant fractions from Ophioglossum thermale were extracted with five different polar solvents using a Soxhlet type extractor. The total phenolic content of the extracts was determined by the Folin-Ciocalteu method. The ethyl acetate fraction of O. thermale was found to contain maximum phenolics. The dried fractions were screened for their antioxidant activity potential using in vitro model systems such as 1,1-diphenyl-2-picryl hydrazyl (DPPH), nitroblue tetrazolium (NBT) and lipid-peroxidation reduction at different concentrations. Results revealed that the EtOAc fraction exhibited the best performance in the DPPH assay, NBT assay and lipid peroxidation. All fractions showed more potent antioxidant capacity than green tea extract, a well-known antioxidant. Furthermore, the EtOAc fraction has the highest total phenolic content (475.65 mg of EGCG/g). In addition, the EtOAc fraction at 0.005% and 0.01% (g/100 ml) also significantly inhibited UVB irradiation-induced ROS generation in human dermal fibroblasts (HDFs). In a carrageenan-induced edema model, the EtOAc fraction showed an inhibitory effect (21.5%, p < 0.05) at 200 mg/kg (p.o.) after 300 min administration. Consequently, 3-O-methylquercetin (3MQ) was also isolated from the antioxidative EtOAc fraction. The data obtained using the above in vitro and in vivo tests suggest that the antioxidant activity of O. thermale and its anti-inflammatory effect on carrageenan-induced acute inflammation can be attributed to its ameliorating effect on oxidative damage, and thus it has great potential as a source for natural health products. To the best of our knowledge, this is the first report on the antioxidant activity of different polar extracts from O. thermale.

Protective effects of fermented Citrus unshiu peel extract against ultraviolet-A-induced photoageing in human dermal fibrobolasts.

The aqueous extracts of Citrus unshiu peel containing flavonoid glycosides was used as co-substrate with Schizophyllum commune mycelia producing ß-glucosidase and its biological activities were studied. ß-glucosidase-produced S. commune mycelia converted the glycosides (narirutin and hesperidin) into aglycones (naringenin and hesperetin). The photoprotective potential of fermented C. unshiu peel extract with S. commune (S-CPE) was tested in human dermal fibroblasts (HDFs) exposed to UVA. It was revealed that S-CPE had an inhibitory effect on human interstitial collagenase (matrix metalloproteinase, MMP-1) expression in UVA-irradiated HDFs. The treatment of UVA-irradiated HDFs with S-CPE resulted in a dose-dependent decrease in the expression level of MMP-1 mRNA. The UVA irradiation raised the proportion of senescence-associated ß-galactosidase (SA-ß-gal) positive cells in comparison with the normal control group. The treatment of UVA-irradiated HDFs with S-CPE was shown to decrease the level of SA-ß-gal (by approximately 45% at an S-CPE concentration 0.1%, w/v) compared with the UVA-irradiated HDFs. It was found that S-CPE containing hesperetin has notable collagen biosynthetic activity for fibroblasts, indicating that S-CPE can be promising cosmetic ingredients.

Biological activities of Fructus arctii fermented with the basidiomycete Grifola frondosa.

Fructus arctii extract containing phenolic glycosides was cultured with Grifola frondosa mycelia to produce ß-glucosidase and its biological activities were studied. This ß-glucosidase converted the glycosides (arctiin and caffeic acid derivatives) into aglycones (arctigenin and caffeic acid). Fermented Fructus arctii extract (G-FAE) with G. frondosa had antioxidant and 5-lipoxygenase inhibitory activities. The photoprotective potential of G-FAE was tested in human dermal fibroblasts (HDF) exposed to ultra-violet A (UVA). It was revealed that G-FAE had an inhibitory effect on human interstitial collagenase (matrix metalloproteinase, MMP-1) expression in UVA-irradiated HDF. The treatment of UVA-irradiated HDF with G-FAE resulted in a dose-dependent decrease in the expression level of MMP-1 mRNA. G-FAE also showed notable stimulation of collagen biosynthetic activity for fibroblasts. These diverse functionalities suggest that G-FAE could be a promising cosmetic ingredient.

Black rice (Oryza sativa L. var. japonica) hydrolyzed peptides induce expression of hyaluronan synthase 2 gene in HaCaT keratinocytes.

Black rice (Oryza sativa L. var. japonica) has been used in folk medicine in Asia. To understand the effects of black rice hydrolyzed peptides (BRP) from germinated black rice, we assessed the expression levels of about 20,000 transcripts in BRP-treated HaCaT keratinocytes using human 1A oligo microarray analysis. As a result, the BRP treatment showed a differential expression ratio of more than 2-fold: 745 were activated and 1,011 were repressed. One of the most interesting findings was a 2-fold increase in hyaluronan synthase 2 (HAS2) gene expression by BRP. Semiquantitative RT-PCR showed that BRP increased HAS2 mRNA in dose-dependent manners. ELISA showed that BRP effectively increased hyaluronan (HA) production in HaCaT keratinocytes.

Anti-oxidative and photo-protective effects of coumarins isolated from Fraxinus chinensis.

Free radicals and reactive oxygen species (ROS), which are generated by UV irradiation, may cause serious injury to skin cell membranes, DNA and functional proteins. In addition, these agents stimulate the expressions of matrix metalloproteinases (MMPs), which can degrade most components of the extracellular matrix (ECM), including collagen. In order to develop new anti-photoaging agents, five major components from the extract of Fraxinus chinensis extract (FCE) were identified. Two of the major components of FCE were found to be esculin (11.2%) and esculetin (1.9%). FCE (IC50: 50.0 microg/mL 1, 1-diphenyl-2-picrylhydrazyl (DPPH); 19.8 microg/mL, superoxide anion radical) and esculetin (IC50: 2.1 microg/mL DPPH; 0.6 microg/mL, superoxide anion radical) showed strong antioxidative activities. Of the compounds tested, esculetin showed the strongest scavenging activity against DPPH radicals, followed by superoxide anions from the xanthine/xanthine oxidase system. The intracellular ROS scavenging activity showed that oxidation of 5-(6-)-chloromethyl-2', 7'-dichlorodihydrofluorescein diacetate (CM-H2DCFDA) was effectively inhibited by esculetin, with potent free radical scavenging activity was also shown in UVB-irradiated human dermal fibroblasts (HDFs). Moreover, treatment of UVA-irradiated HDFs with esculetin resulted in dose-dependent decreases in the expression levels of MMP-1 mRNA and protein. From these results, FCE and one of its components, esculetin, were predicted to be potentially useful as ingredients in cosmetics for protecting against photoaging.

Antioxidative activity of the hydrolytic enzyme treated Sorbus commixta Hedl. and its inhibitory effect on matrix metalloproteinase-1 in UV irradiated human dermal fibroblasts.

Superoxide radical scavenging activity and DPPH radical scavenging activity were assessed in order to evaluate the antioxidant effect of the Sorbus commixta Hedl. extract (SCoE). SCoE was also treated with several carbohydrate-hydrolytic enzymes that significantly increased the total phenol and flavonoid composition of SCoE. The enzymatically treated SCoE was then assessed for antioxidative activity. The most efficient radical scavenging activity was observed when SCoE was treated with -glucanase. The radical scavenging activity of beta-glucanase-treated SCoE (beta-GSCoE) enhanced the viability of human dermal fibroblasts (HDFs) exposed to ultraviolet (UV) light. The intracellular reactive oxygen species (ROS) scavenging activity of beta-GSCoE was assessed using UVB (20 mJ/cm2)-irradiated HDFs. UVB irradiation increased dichlorofluorescein (DCF) fluorescence, which was measured by a 5-(6-)chloromethyl-2',7'- dichlorodihydrofluorescein diacetate (CM-H2DCFDA). DCF-fluorescence was significantly decreased in the beta-GSCoE-containing culture medium, suggesting that beta-GSCoE scavenges free radicals. The protective effect was further verified by assessing the expression of matrix metalloproteinase-1 (MMP-1) in UVA-irradiated HDFs. The treatment of UVA-irradiated HDFs with beta-GSCoE resulted in a dose-dependent decrease in the expression level of MMP-1 protein and mRNA. These results suggest that beta-GSCoE may mitigate the effects of photoaging in skin by reducing UV-induced adverse skin reactions.

Antioxidative and acute antiinflammatory effects of Torreya grandis.

The present study was undertaken to investigate the antioxidative and antiinflammatory activities of the ethanolic extract of seeds of Torreya grandis (EST). Exposure of human dermal fibroblasts to the extract at 50 and 250 microg/ml showed significant protective effect against hydrogen peroxide (300 microM). EST not only protected cell survival from H(2)O(2)-induced toxicity, but also inhibited the H(2)O(2)-induced LDH release significantly. It was also found that EST at 100 and 1000 microg/ml showed scavenging activities of radicals and reactive oxygen species with 29.8% and 100.0% of inhibition against DPPH radical and 41.2% and 98.4% against superoxide radicals in the xanthine/xanthine oxidase system, respectively. Topically applied EST dose-dependently inhibited arachidonic acid (AA)- and 12-O-tetradecanoylphorbol-13-acetate (TPA)-induced ear edema in mice.

Antioxidative and acute anti-inflammatory effects of Campsis grandiflora flower.

The present study was undertaken to investigate the antioxidative and anti-inflammatory activities of the extract of the flower of Campsis grandiflora (Thunb.) K. Schum. Exposure of human dermal fibroblasts to 50% EtOH extract of Campsis grandiflora flower (ECG) at 10 and 100 microg/ml showed significant protective effect against hydrogen peroxide (300 microM). ECG not only protected cell survival from H(2)O(2)-induced toxicity, but also inhibited the H(2)O(2)-induced leakage of lactate dehydrogenase (LDH) enzyme release and DNA fragmentation significantly. It was also found that ECG showed scavenging activities of radicals and reactive oxygen species with IC(50) values of 20 microg/ml against 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical and 52 microg/ml against superoxide radicals in the xanthine/xanthine oxidase system, respectively. Topically applied ECG dose-dependently inhibited arachidonic acid (AA)- and 12-O-tetradecanoylphorbol 13-acetate (TPA)-induced ear edema in mice. Consistent with its antioxidative properties in vitro, the present results suggest the therapeutic potential of ECG for acute skin inflammation that may involve oxidative tissue damage.

Stimulative and sedative effects of essential oils upon inhalation in mice.

This study investigated the stimulative or sedative effects of inhaling fragrant essential oils (EOs) by using a forced swimming test (FST) with mice. This behavioral test is commonly used to measure the effects of antidepressant drugs. The inhalation by mice of EOs, such as ginger oil (p<0.05), thyme oil (p<0.05), peppermint oil (p<0.05), and cypress oil (p<0.01) resulted in 5% to 22% reduction of immobility. The same results were achieved when over-agitation was artificially induced in the mice by an intraperitoneal injection of caffeine (a psycho-stimulant). In contrast, inhalation of some EOs by the mice resulted in increased immobility. To evaluate more correctly the sedative effects of EOs, the immobility of over-agitated mice induced with caffeine was ascertained after the inhalation of various EOs. Inhalation of lavender oil (p<0.01) and hyssop oil (p<0.01) increased the immobile state in mice that were treated with caffeine. The results of this study indicate that the inhalation of essential oils may induce stimulative or sedative effects in mice.