Anti-Inflammatory, Barrier-Protective, and Antiwrinkle Properties of Agastache rugosa Kuntze in Human Epidermal Keratinocytes.

This work aimed to assess the skin-beneficial properties of Agastache rugosa Kuntze, an herbal medication used to treat different types of disorders in traditional folk medicine. The total phenolic compounds and total antiradical, nitrite scavenging, superoxide scavenging, antielastase, and antihyaluronidase activities of a hot water extract of A. rugosa Kuntze leaves (ARE) were spectrophotometrically determined. Intracellular reactive oxygen species (ROS) was fluorometrically quantitated using 2',7'-dichlorodihydrofluorescein diacetate (DCFH-DA). Inducible nitric oxide synthase (iNOS) and filaggrin were evaluated using Western analysis. Real-time quantitative RT-PCR was used to measure filaggrin mRNA. Caspase-14 activity was determined using a fluorogenic substrate. ARE contained the total phenolic content of 38.9 mg gallic acid equivalent/g extract and exhibited 2,2'-diphenyl-1-picrylhydrazyl (DPPH) radical, superoxide radical, and nitrite scavenging activities with the SC50 values of 2.9, 1.4, and 1.7 mg/mL, respectively. ARE exerted suppressive activities on nitric oxide (NO) and ROS levels elevated by lipopolysaccharide (LPS) or tumor necrosis factor-α (TNF-α) in HaCaT keratinocytes. It attenuated the LPS-stimulated expression of iNOS. ARE augmented the UV-B-reduced filaggrin expression on both protein and mRNA levels and was capable of upregulating the UV-B-reduced caspase-14 activity. ARE inhibited in vitro elastase and hyaluronidase activities associated with the wrinkling process. ARE, at the concentrations used, did not interfere with the viability of HaCaT keratinocytes. These findings preliminarily imply that the leaves of A. rugosa possess desirable cosmetic potentials, such as anti-inflammatory, barrier protective, and antiwrinkle activities, which infers their skin healing potentials.

Probiotic fermentation augments the skin anti-photoaging properties of Agastache rugosa through up-regulating antioxidant components in UV-B-irradiated HaCaT keratinocytes.

BACKGROUND: Agastache rugosa (Fisch. & C.A.Mey.) Kuntze (Korean mint) is used to treat diverse types of human disorders in traditional medicine. In recent years, its non-fermented leaf extract (ARE) has been shown to possess protective properties against ultraviolet-B (UV-B) radiation-induced photooxidative stress. The present work aimed to examine whether probiotic bacterial fermentation would potentiate the skin anti-photoaging activity of ARE or not, by comparing the protective properties of ARE and corresponding fermented extract (ARE-F) against UV-B radiation-induced photooxidative stress in HaCaT keratinocytes. METHODS: ARE-F was produced from ARE by the fermentation with Lactobacillus rhamnosus HK-9, a type of Gram-positive probiotic bacterial strain. Anti-photoaging activities were evaluated by analyzing reactive oxygen species (ROS), promatrix metalloproteinases (proMMPs), total glutathione (GSH) and total superoxide dismutase (SOD) in UV-B-irradiated HaCaT keratinocytes. Antiradical activity was determined using 2,2-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) radical scavenging assay. RESULTS: ARE-F contained higher attenuating activity on the UV-B-induced ROS generation than ARE. Similarly, ARE-F was able to diminish the UV-B-induced proMMP-9 and -2 more effectively than ARE. ARE-F displayed higher tendencies to augment the UV-B-reduced total GSH content and SOD activity than ARE. However, there were no significant difference between ARE and ARE-F in ABTS radical scavenging activities. CONCLUSIONS: The findings suggest that the UV-B radiation-protective activity of ARE is enhanced by probiotic bacterial fermentation, which might improve the therapeutic and cosmetic values of A. rugosa leaves.

Protective properties of geniposide against UV-B-induced photooxidative stress in human dermal fibroblasts.

CONTEXT: Geniposide (genipin-1-O-ß-d-glucoside) is a major bioactive ingredient in the fruits of gardenia [Gardenia jasminoides J. Ellis (Rubiaceae)], a traditional herbal medicine in Asian countries. OBJECTIVE: This work assesses the skin anti-photoaging potential of geniposide in human dermal fibroblasts under UV-B irradiation. MATERIALS AND METHODS: The anti-photoaging property of geniposide, at varying concentrations (5, 12 and 30 µM) treated for 30 min prior to UV-B irradiation, was evaluated by analysing reactive oxygen species (ROS), promatrix metalloproteinase-2 (proMMP-2), glutathione (GSH), superoxide dismutase (SOD), nuclear factor erythroid 2-related factor 2 (Nrf2) and cellular viability. RESULTS: Geniposide suppressed the ROS elevation under UV-B irradiation, which was revealed using three ROS-sensitive fluorescent dyes. The use of 2',7'-dichlorodihydrofluorescein diacetate (DCFH-DA), dihydroethidium (DHE) and dihydrorhodamine 123 (DHR-123) elicited the IC50 values of 10.5, 9.8 and 21.0 µM, respectively. Geniposide attenuated proMMP-2 at activity and protein levels that were elevated under UV-B-irradiation. Geniposide at 5, 12 and 30 µM augmented the UV-B-reduced total GSH content to 1.9 ± 0.1-, 2.2 ± 0.2- and 4.1 ± 0.2-fold, respectively. Geniposide at 5, 12 and 30 µM upregulated total SOD activity to 2.3 ± 0.1-, 2.5 ± 0.3- and 3.3 ± 0.3-fold, respectively, under UV-B irradiation. The UV-B-reduced Nrf2 levels were also upregulated by geniposide treatment. Geniposide, at the concentrations used, was unable to interfere with cellular viabilities under UV-B irradiation. DISCUSSION AND CONCLUSIONS: After the skin anti-photoaging potential of geniposide may be further verified, it can be utilized as a safer resource in the manufacture of effective anti-aging cosmetics.

Attenuating properties of Agastache rugosa leaf extract against ultraviolet-B-induced photoaging via up-regulating glutathione and superoxide dismutase in a human keratinocyte cell line.

Agastache rugosa Kuntze, known as a Korean mint, is an herbal medicine that has been used for the treatment of diverse kinds of symptoms in traditional medicine. This work was undertaken to assess the protective properties of A. rugosa leaves against UV-B-induced photoaging in HaCaT keratinocytes. They were evaluated via analyzing reactive oxygen species (ROS), promatrix metalloproteinase-2 (proMMP-2) and -9 (proMMP-9), total glutathione (GSH), total superoxide dismutase (SOD), cellular viability, flavonoid content and in vitro radical scavenging activity. Total flavonoid content of ARE, a hot water extract of A. rugosa leaves, was 22.8±7.6mg of naringin equivalent/g ARE. ARE exhibited ABTS(+) radical scavenging activity with an SC50 of 836.9µg/mL. ARE attenuated the UV-B-induced ROS generation. It diminished the UV-B-induced elevation of proMMP-2 and -9 at both activity and protein levels. On the contrary, ARE was able to enhance the UV-B-reduced total GSH and total SOD activity levels. ARE, at the used concentrations, was unable to interfere with the cellular viabilities of HaCaT keratinocytes under UV-B irradiation. Taken together, ARE possesses a protective potential against UV-B-induced photoaging in HaCaT keratinocytes, possibly based upon up-regulating antioxidant components, including total GSH and SOD. These findings reasonably suggest the use of A. rugosa leaves as a photoprotective resource in manufacturing functional cosmetics.

Contribution of ginsenoside Re to cellular redox homeostasis via upregulating glutathione and superoxide dismutase in HaCaT keratinocytes under normal conditions.

Ginsenoside Re (Re) is one of the main ginsenosides which are known to be responsible for diverse pharmacological properties of ginseng, widely used as a dietary supplement and a general tonic. The present work was undertaken to evaluate the antioxidative property of Re by analyzing reactive oxygen species (ROS), nitric oxide (NO), pro-matrix metalloproteinase-2 (proMMP-2) and -9 (proMMP-9), total glutathione (GSH) and superoxide dismutase (SOD) in normal, unstressed HaCaT keratinocytes. When HaCaT cells were subjected to Re, Re suppressed the ROS and NO levels in a concentration-dependent manner. Re at concentrations used exhibited no cytotoxicity on the cellular viabilities of HaCaT cells. It was also able to attenuate proMMP-2 and -9 at both activity and protein levels. On the contrary, Re was capable of enhancing the total GSH and SOD activity levels. The findings suggest that Re has an antioxidative property through the upregulation of some antioxidant components, including total GSH and SOD, in HaCaT keratinocytes, which then can play its underlying role in maintaining the cellular redox homeostasis.

Effect of balanced low pressure drying of curcuma longa leaf on skin immune activation activities.

The effect of balanced low pressure drying pretreatment associated with ultrasonication extraction (BU) on the enhancement of skin immune modulatory activities of Curcuma longa leaf was studied by comparing with conventional hot air drying (HE), freeze drying (FE) and balanced low pressure drying (BE) pretreatment processes. In considering skin immune activation activities such as the inhibition of hyaluronidase activity, the BU extract showed ca. 10% higher than those of HE, and even higher than that of the FE extract. Nitric oxide production from macrophage of the BU extract in adding 1.0 mg/mL was increased up to 16.5 µM. When measuring inhibition of IL-6 and TNF-a production from the human T lymphocytes (T cell), the BU extract also showed 53% and 78% of inhibition effect, respectively. It is found that the BU extract could effectively suppress the expression levels of skin inflammation related genes such as Cox-2 and iNOS, down to 80% and 85% compared to the control, respectively. Balanced low pressure drying process was especially active on dehydration of the leaves with minimizing the destruction and making easier elution of the bioactive substances, which resulted in higher extraction yield and better biological activities.

Anti-Inflammatory, Antioxidant, Anti-Angiogenic and Skin Whitening Activities of Phryma leptostachya var. asiatica Hara Extract.

This work aimed to assess some pharmacological activities of P. leptostachya var. asiatica Hara. The dried roots of P. leptostachya var. asiatica Hara were extracted with 70% ethanol to generate the powdered extract, named PLE. Anti-angiogenic activity was detected using chick chorioallantoic membrane (CAM) assay. In vitro anti-inflammatory activity was evaluated via analyzing nitric oxide (NO) content, inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) in lipopolysaccharide (LPS)-stimulated RAW264.7 macrophage cells. Antioxidant activity was determined by 1,1-diphenyl-2-picrylhydrazyl (DPPH) assay and reactive oxygen species (ROS) level in the stimulated macrophage cells. Matrix metalloproteinase-9 (MMP-9) and -2 (MMP-2) activities in the culture media were detected using zymography. PLE exhibits an anti-angiogenic activity in the CAM assay, and displays an inhibitory action on the generation of NO in the LPS-stimulated macrophage cells. In the stimulated macrophage cells, it is able to diminish the enhanced ROS level. It can potently scavenge the stable DPPH free radical. It suppresses the induction of iNOS and COX-2 and the enhanced MMP-9 activity in the stimulated macrophage cells. Both monooxygenase and oxidase activities of tyrosinase were strongly inhibited by PLE. Taken together, the dried roots of P. leptostachya var. asiatica Hara possess anti-angiogenic, anti-inflammatory, antioxidant and skin whitening activities, which might partly provide its therapeutic efficacy in traditional medicine.

Potentiation of antioxidative and anti-inflammatory properties of cultured wild ginseng root extract through probiotic fermentation.

OBJECTIVES: This work aimed to determine some pharmacological properties of non-fermented (WG) and fermented (FWG) extracts of cultured wild ginseng root. METHODS: WG was treated with Bifidobacterium longum to generate FWG. Ginsenoside patterns were analysed using thin-layer chromatography and high-performance liquid chromatography. The effect of WG and FWG on reactive oxygen species (ROS) was examined in lipopolysaccharide-stimulated RAW264.7 macrophage cells. Intracellular ROS were detected by flow cytometry. Nitrite in culture supernatant fractions was determined using the Griess reaction. 1,1-Diphenyl-2-picrylhydrazyl was used to determine anti-radical activity. Cell viability was determined using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay. KEY FINDINGS: FWG was rich in ginsenosides Rg3 and Rh2, compared with WG. FWG diminished the enhanced ROS level more strongly than WG in lipopolysaccharide-stimulated RAW264.7 macrophage cells. Both WG and FWG decreased the nitrite levels in stimulated macrophage cells with half-maximal inhibitory concentration (IC50) values of 2.7 and 1.5 mg/ml, respectively, implying that FWG had an enhanced anti-inflammatory activity. Neither WG nor FWG exhibited cytotoxicity on the macrophage cells. In the radical scavenging assay, the IC50 values of WG and FWG were 32.6 and 0.78 mg/ml, respectively, suggesting that FWG had an increased scavenging activity. CONCLUSIONS: FWG possesses enhanced antioxidative and anti-inflammatory activity, indicating that fermentation of cultured wild ginseng root extract with a probiotic bacterium can strengthen some of its desirable effects.

The effect of ultrasonication on the immunomodulatory activity of low-quality ginseng.

This study investigated the effects of ultrasonication extraction (UE) on the immunomodulatory activity of low-quality ginseng. The results indicate that the optimal conditions for extracting low-quality ginseng are ultrasonication at 60 kHz and 85°C for 60 min. The extraction yield from the UE was 20% higher than that of the water extraction (WE) at 100°C. The low quality ginseng obtained from the UE exhibited relatively low cytotoxicity toward normal human cells, with an observed toxicity of 15-18% at a concentration of 1.0 mg/mL. The ginseng product obtained following UE induced human B and T cells growth and resulted in concentrations of up to 9.33 × 10(4) cells/mL and 15.33 × 10(4) cells/mL, respectively. The ginseng extract also increased the secretion of interleukin-6 and tumor necrosis factor-α from these cells by up to 35%, and natural killer/ cell growth was also improved by up to 30%. The UE effectively released 2- to 3-fold higher levels of ginsenosides than the WE. Specifically, the obtained levels of Rb(1) , Re, and Rg(1) , which are likely immunomodulatory factors, were approximately three times higher after ultrasonication than after WE. These results were further supported by the finding that UE product-treated macrophages produced higher levels of nitric oxide (21 µM) than macrophages treated with the WE product or with standard ginsenosides. These results demonstrate that this optimized ultrasonication process effectively destroyed the more rigid cell walls of low-quality ginseng and released high levels of ginsenosides. This work is the first to correlate extraction parameters with both extraction yields and biological activity. The use of low-quality ginseng can thus be expanded by utilizing a low-temperature ultrasonic extraction process.

Enhancement of anti-inflammatory and antinociceptive actions of red ginseng extract by fermentation.

OBJECTIVES: This work aimed to compare some pharmacological properties of red ginseng extract (RG) and fermented red ginseng extract (FRG). METHODS: Antinociceptive activity was analysed using the acetic acid-induced abdominal constriction response. Anti-inflammatory activity was evaluated using acetic acid-induced vascular permeability and carrageenan-induced inflammation in the air pouch, and analysed through the measurement of nitrite content in the lipopolysaccharide (LPS)-stimulated macrophage cells. Anti-angiogenic activity was determined using the chick chorioallantoic membrane assay. KEY FINDINGS: In-vivo anti-inflammatory activity of FRG was stronger than that of RG in two animal models, vascular permeability and air-pouch models. In the vascular permeability model, the doses of RG and FRG required for half-maximal inhibition (IC50) were 181 and 59mg/kg, respectively. FRG exhibited significantly stronger antinociceptive activity than RG. In the acetic acid-induced abdominal constriction response, the IC50 values of RG and FRG were 153 and 27mg/kg, respectively. Although both RG and FRG were able to suppress production of nitric oxide in the LPS-stimulated RAW264.7 macrophage cells, the suppressive activity of FRG appeared to be stronger than that of RG. However, RG and FRG showed similar anti-angiogenic activity. CONCLUSIONS: FRG possesses enhanced anti-inflammatory and antinociceptive activity but similar anti-angiogenic activity than RG.

Enhancement of the skin-protective activities of Centella asiatica L. Urban by a nano-encapsulation process.

Aqueous extracts of Centella asiatica L. Urban were encapsulated by an edible biopolymer, gelatin, which has no effect on their cosmetic activities. The nanoparticles were w/o-type spherical liposomes that had an average diameter of 115.0nm. The encapsulation efficiency was estimated to be approximately 67%, which was relatively high for these aqueous extracts. The nanoparticles showed lower cytotoxicity (10%) in human skin fibroblast cells than the unencapsulated crude extract (15%) at 1.0mg/ml, this was possibly because a smaller amount of the extract was present in the nanoparticles. The nanoparticles efficiently reduced the expression of matrix metalloproteinase (MMP)-1 in UV-irradiated cells from 136.1% to 77.6% (UV-irradiated control) and inhibited hyaluronidase expression (>60%) at a concentration of 0.5mg/ml, which was higher than the levels produced by the unencapsulated crude extracts. The nanoparticles had a very high flux through mouse skin and also remained at relatively large concentrations in the derma when compared to the unencapsulated crude extracts. These results clearly indicate that the skin-protective activities of C. asiatica were significantly improved through the nano-encapsulation process. These findings also imply that a crude extract can be used and have the same efficacy as purified compounds, which should reduce the purification process and production costs.