In order to investigate the optimal UVB (ultraviolet B) treatment conditions for vitamin D2 enhancement of freeze-dried ear mushroom, sample size (below 300 μm~whole), UV treatment temperature (30~60℃), treatment density (6.25~50.0 mg/cm2) and the samples mixing frequency (1~32 times) were treated differently. After that, chromaticity, vitamin D2 and ergosterol (vitamin D2 precursor) contents were investigated. As a result of the investigating, effective UVB treatment conditions for vitamin D2 enhancement are as follows. The sample sizes were 2~4 mm and finely crushed pieces. The treatment temperatures were 50℃ and 60℃. The treatment density was 12.5 to 25.0 mg/cm2, and the number of sample mixing was 8 times or more. As the amount of vitamin D2 increased by UVB treatment, the ergosterol content generally tended to decrease. However, under some UVB treatment conditions, the vitamin D2 content was not high despite the decrease of ergosterol content. Under the conditions set in this experiment, it was possible to obtain ear mushrooms with enhanced vitamin D2 up to 26,968.7 μg/100 g. Therefore, it is thought that the ear mushroom is highly likely to be used as a vitamin D source and nutritionally fortified food ingredient.

Repetitive or excessive exposure to ultraviolet (UV) radiation causes oxidative stress-mediated skin photoaging through the overproduction of reactive oxygen species. Actinidia polygama is known as a medical plant used in oriental medicine for treating several diseases such as abdominal pain, stroke and rheumatoid arthritis. Recently, it was reported that A. polygama extract had anti-wrinkle and skin hydrating properties in ultraviolet B (UVB)-exposed hairless mice. However, the molecular biological mechanism of this extract on alleviating skin photoaging is still unknown. Therefore, we investigated the anti-photoaging effects of PB203, which is the powder of A. polygama extract, in the in vivo and in vitro photoaging models. First, PB203 showed 2,2’-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) and 2,2-diphenyl-1-picrylhydrazyl radical scavenging activities due to the presence of anti-oxidant components including flavonoids and polyphenols. In UVB-irradiated hairless mice, oral administration of PB203 (100 mg/ kg) significantly improved wrinkle formation, skin dehydration, elasticity and skin barrier function by decreasing the levels of matrix metalloproteinases (MMPs) and increasing those of collagen I, filaggrin, involucrin and loricrin. Especially, the reduced production of p-p38, p-c-Jun and p-c-Fos by PB203 reversed the elevated levels of MMPs mediated by UVB exposure, resulting in the upregulation of collagen I expression. Consistent with these animal data, PB203 remarkably enhanced the mRNA expression of collagen I, filaggrin, involucrin and loricrin, while suppressed that of MMPs in UVB-irradiated HaCaT cells. And PB203 increased the wound recovery rate of cells by promoting their proliferation and migration. Moreover, PB203 significantly recovered the activity of superoxide dismutase inhibited by UVB in both mice and cells. In conclusion, PB203, which protects skin from UVB-induced photodamage by exerting antioxidant properties, can be considered to have sufficient potential as a functional ingredient or therapeutic agent improving skin photoaging and related skin symptoms.

피부는 인체를 구성하는 가장 큰 장기로 생체 내부를 보호한다. 자외선은 피부에 광노화와 산화 적 손상을 비롯한 다양한 염증반응을 일으킨다. 본 연구의 목적은 섬유아세포에서 UVB를 조사하여 Saponaria 추출물의 보호 효과를 조사하는 것이다. 본 연구에서는 UVB에 의한 세포독성과 산화적 세포사 멸, NO 및 PGE2 생성에 대한 보호활성을 나타내는 Saponaria의 유효성을 평가하였다. HS68 세포를 UVB(120mJ/cm2)에 조사하고 100, 200, 400 μg/mL의 다양한 농도로 Saponaria 추출물로 24시간 동안 처리하였으며, 자외선 B에 의해 생성된 세포 내 활성 산소 종(ROS)은 DCF-DA 염색 후 분광 형광계를 사용하여 검출하였다. 또한 지질 과산화는 배양 배지로 분비되는 8-이소프로스탄의 수준을 측정하여 분석 하였다. 그 결과 Saponaria 추출물이 UVB에 의한 세포독성을 효과적으로 억제하였다. 산화적 세포 손상은 UVB로 유도된 HS68 섬유아세포에서 PGE2를 매개하였고, 이는 사포나리아 추출물 처리에 의하여 유의하 게 억제되었다. 또한, 이들 추출물의 보호 효과는 농도 의존적으로 세포내 ROS 생성 및 지질 과산화 억제 에 의해 매개되는 것으로 평가되었다. 이러한 결과는 Saponaria 추출물이 자외선 B에 의한 산화적 스트레 스로 매개한 피부 손상을 억제하여 세포 보호효과를 나타내므로 항노화 기능성 소재로 활용될 수 있을 것 으로 사료된다.

We evaluated the protective effects of cricket methanol extract (CME) on ultra-violet B (UVB)-induced photoaging in human skin fibroblasts. The fibroblast cells were treated with 10, 50, and 100 μg/mL of CME for 24 h, and then exposed to UVB (30 mJ/cm2). CME showed a dose-dependent cytoprotective effect without any observable cytotoxicity. CME reduced UVB-induced production of reactive oxygen species (ROS) by 34.4, 34.9, 40.6% at concentrations of 10, 50, 100 μg/mL respectively. CME inhibited the release of matrix metalloproteinase (MMP) 1 and 3. Furthermore, CME also reduced UVB-induced collagen degradation in the fibroblast cells. Taken together, our data suggests that CME has a significant protective effect on UVB-induced photoaging of the skin. This benefit occurs through multiple mechanisms. The results also suggest a potential role for CME as an ingredient in anti-photoaging cosmetic products in the future.

지구상에서 가장 오래된 해양 미세조류로 알려진 스피룰리나는 인체에 필요한 영양성분을 대부분 함유하고 있는 것으로 알려져 있다. 그 구성성분으로는 phycocyanin, chlorophyll, β-carotene과 같 은 다양한 물질을 다량 함유한다고 보고되고 있으며, 노화 및 미백효과가 있다고 알려져 있다. 본 연구에서 는 스피룰리나 정제수 추출물의 UVB로 유도된 활성산소종 ROS (reactive oxygen species) 소거능과 항산 화능을 확인하였다. 스피룰리나 정제수 추출물 0.05, 0.10, 0.50 1.0 mg/mL의 DPPH 라디칼 소거능, FRAP 환원능 및 ABTS+ 라디칼 소거능을 측정하여 항산화효과를 확인하였다. 대체실험동물모델인 Zebrafish를 이용하여 스피룰리나 정제수 추출물을 0.05, 0.10, 0.50 mg/mL 농도로 처리하여 응고율, 부 화율, 심장독성을 측정하였으며, 스피룰라나 추출물을 0.05, 0.10, 0.50 mg/mL 농도로 처리한 후, DCFH-DA로 염색하여 UVB로 유도된 ROS 저해효과를 확인하였다. 항산화효과 측정 결과 양성대조군인 ascorbic acid와 비교시 DPPH, Frpa, ABTS 모두 농도 의존적으로 항산화 효과가 있음을 확인하였다. 대체 실험동물인 Zebrafish를 이용하여 응고율과 부화율, 심박수를 측정한 결과 0.5 mg/mL을 제외한 0.05, 0.10 mg/mL에서는 대조군과 비교 시 독성이 없음을 확인 하였다. UVB로 유도된 Zebrafish의 ROS 소거 능은 양성대조군에 비해 높은 ROS 감소 효과를 나타내었다. 본 연구의 결과는 스피룰라나 정제수 추출물 이 자외선 및 피부 보호 화장품 소재로 사용가치가 있음을 시사한다.

To examine the possibility of ear mushroom (EM) as a source of natural vitamin D, the UVB (ultraviolet B) was treated according to sample drying status, drying methods before UVB treatment and harvest time. And then, vitamin D2 and ergosterol contents were investigated. According to the sample drying status, the vitamin D2 contents of fresh and freeze-dried EM (whole) increased to 4,634.4~4,780.9 μg/100 g D.W. (dry weight) under UVB dose 52.5~70.0 kJ/m2 and above 18,693.1 μg/100 g D.W. under above 105 kJ/m2, respectively. By drying methods before UVB treatment, vitamin D2 contents of EM powder (below 500 μm) that dried in the vinyl house and freeze-dryer increased to 4,886.2~5,132.9 μg/100 g D.W. under above 105 kJ/m2 and 17,103.7 μg/100 g D.W. under 70 kJ/m2, respectively. Ergosterol content decreased with increasing UVB dose in all experiments. According to the harvest time, vitamin D2 content under UVB dose 210 kJ/m2 showed marked difference and in order of June, July, August, October and April. As for the results, the optimum harvest time, drying method before UVB treatment, sample size, UVB dose for the EM contained high vitamin D2 content were June, freeze-drying, whole, and 105 kJ/m2, respectively.

Mature silkworm of Bombyx mori is known to contain various functional materials. However, it is too hard to chew or digest for humans when it is cooked or lyophilized. In Korea, the Rural Development Administration recently developed and patented a method for making mature silkworms edible. In this study, therefore, we examined the effects of a larval powder of steamed mature silkworm (SMSP) on skin pigmentation and melanogenesis. For elucidating the depigmenting activity, lightness of a designated site on the murine dorsal skin was measured in vivo. During the experiment, hyperpigmentation was induced on the skin by cumulative exposure to ultraviolet B (UVB) irradiation. At the end of the experiment, melanin production on the skin was visualized by Fontana-Masson staining. Orally administered SMSP of pistachio cocoon strain for at least a month or longer significantly and reduced abnormal pigmentation caused by UVB on the murine dorsal skin. SMSP also showed a potential anti-melanogenic efficacy in modulating UVB-induced hyperpigmentation. Taken together, SMSP was identified as a potential candidate for a novel anti-melanogenic agent, which showed depigmenting efficacy against UVB-induced hyperpigmentation in vivo when administered orally.

Ultraviolet B (UVB) exposure is a risk factor for skin damage resulting in oxidative stress, inflammation, and cell death. The purpose of this study was to investigate the physicochemical properties of Platycodon grandiflorum (PG) to improve its biological activities using a three-step steaming process. We investigated the protective effects of PG and steamed PG extracts on human dermal fibroblasts (HDFs) against UVB radiation-induced oxidative stress and inflammation as well as the underlying mechanisms. The antioxidant potential of the PG extracts was evaluated by measuring the 2,2-diphenyl-1- picrylhydrazyl (DPPH) and 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid (ABTS) scavenging activity. ABTS and DPPH were shown by the 0, 30, and 70% ethanol extracts of 2S-PG and 3S-PG (IC50, 28~45 and 27~30 μg/mL, respectively). Treatment of UVB-irradiated cells with steamed PG (25~400 μg/mL) did not affect their viability. The streamed PG extract suppressed UVB-induced generation of reactive oxygen species (ROS). In addition, streamed PG extract reduced cyclooxygenase-2 (COX-2) and inducible nitric oxide synthase (iNOS) protein expression in UVB-irradiated HDF, regulating nuclear factor (NF)-κB expression. These findings suggest that steamed PG extract may be potentially effective against inflammation associated with UVB-induced oxidation stress.

스피룰리나는 지구상의 가장 오래된 조류중 하나이며 피코시아닌, 토코페롤, 베타카로틴을 함유하고 있어 노화를 방지한다고 보고된 바 있다. 본 연구에서는 재배방법이 다른 스피룰리나 에탄올추출물의 UVB로 유도된 활성산소종(reactive oxygen species, ROS)의 저해와 항산화효과를 확인하였다. 실내배양 스피룰리나 에탄올추출물(ICAE)과 야외배양 스피룰리나 에탄올추출물(OCAE) 0.1, 0.5, 1 ㎎/㎖의 DPPH 라디칼 소거능과 SOD 유사 활성을 측정하여 항산화효과를 확인하였다. UVB로 유도된 ROS의 저해 효과는 제브라피쉬 배아와 HaCaT 세포를 이용하여 확인하였다. ICAE와 OCAE 0, 0.01, 0.05, 0.1, 0.5, 1 ㎎/㎖을 제브라피쉬 배아와 HaCaT 세포에 처리하고 UVB로 ROS를 유도하였고 DCFH-DA로 염색하여 세포내 ROS의 양을 검출하였다. 항산화효과를 측정한 결과 양성대조군인 ascorbic acid의 DPPH 라디칼 소거능은 73%, SOD 유사 활성은 86%로 나타났다. ICAE와 OCAE 1 ㎎/㎖의 농도에서 각각 43, 57%의 DPPH 라디칼 소거능을 나타내었고, 20, 19%의 SOD 유사 활성을 나타내어 양성대조군인 ascorbic acid보다 낮았지만 유의적인 항산화효과가 있었다. UVB로 유도된 제브라피쉬 배아와 HaCaT 세포의 ROS가 음성대조군보다 증가하였고, ICAE와 OCAE 처리된 군은 농도 의존적으로 UVB만 조사된 양성대조군 보다 ROS가 감소하였다. 본 연구의 결과는 스피룰리나 에탄올추출물이 피부보호용 화장품 소재로 사용가치가 있음을 시사한다.

In the present study, we investigated the protective effect of various grain methanolic extracts against UVB-induced photo-aging in human skin fibroblasts. Various grain methanolic extracts were evaluated for their antioxidant compounds and activities. 2,2-Ddiphenyl-1-picryhydrazyl radical (DPPH) and ABTS 2,2-azino-bris-(3-ethylbenzoth iazoline-6-sulphonic acid) radical cation scavenging activities have been used to measure the relative antioxidant activities of extracts from grains. The content of total polyphenolics in the extracts were evaluated using spectrophotometric methods. Human skin fibroblast (Hs68) cells were pretreated with various grain methanolic extracts (25 μg/mL). Skin toxicity was simulated by exposing the cells to UVB (30 mJ/cm2) irradiation. In response to the UVB-irradiation, an increased amount of matrix metalloproteinases (MMPs) release was observed, whereas pretreatment of various grain methanolic extracts significantly inhibited the production of MMP-1 in Hs68 cells. We also found that pretreatment of the extracts significantly decreased UVB-induced reactive oxygen species and significantly increased total collagen content in Hs68 cells. These results provide that grains could be regarded as a potential ingredient in natural cosmetics, used for UVB protection.

It has been reported that collagen and elastin inhibit skin wrinkle formation. Skin elasticity is closely associated with the formation of wrinkles induced by UV exposure. In this study, we investigated the protective effects Pleurotus ostreatus (P. ostreatus) on UVB irradiation induced wrinkle in hairless mice. We evaluated for their free radical-scavenging activities against 1,1-diphenyl-2-picrylhydrazyl (DPPH), the anti-elastase activities, and for their anti-matrix metalloproteinase-1 (MMP-1) activity in human skin fibroblast cells. In the wrinkle measurement and image analysis of skin replicas, the results showed that P. ostreatus significantly inhibited wrinkle formation caused by chronic UVB irradiation. These results suggest that P. ostreatus has anti-wrinkle activity.

Cycloamyloses (CA) from high-amylose rice starch (Dodamssal, 50.94% amylose) was produced to form a complex with anthocyanin extracted from black rice (sihtoheukmi). The effects of CA concentration (1, 5, 10 and 30%, w/v), pH (2, 4, 6, and 8), UVB irradiation time (~24h) and thermal treatment time (~24h) on the oxidant capacity of the complex were investigated. Anthocyanin consisting of 95% cyanidin-3-glucoside (C3G) was extracted from the black rice by suspending in 60%(v/v) ethanol containing 0.2%(v/v) hydrochloric acid for 90 min at 50°C. Cycloamyloses (CA) produced by 4-α-glucanotransferase (4αGTase) formed complexes with C3G by shaking the mixture for 48 hours at 30°C. Antioxidant activities (radical scavenging capacity) were determined by using ABTS [2,2'-azinobis-(3-ethylbenzothiazoline- 6-sulfonic acid)]. The antioxidant activities of the complexes increased with pH increasing. Upon UVB irradiation and heat treatment, the antioxidant activities were more retained for the complexes incorporating 10% CA concentration or above compared to the C3G control. Degradation rate constant (Kd) and half-life (T1/2) were obtained from degradation data of CA complex with cyanidin-3-glucoside. The effect of CA complex formation with C3G is significantly observed at pH4. The results suggest that the CA complex formed with C3G indeed has an enhanced antioxidant potential toward UVB and thermal degradation.

To increase the stability of anthocyanin from black rice, β-cyclodextrin (β-CD) formed a complex with anthocyanin, and thermal and UVB stability of the complex was investigated at various pH levels. The formation of β-CD complex with anthocyanin was confirmed by the decreased UV-Vis spectrophotometer absorbance (at 511nm) with increasing the concentration of β-CD at pH 2, 2.5, 3, 4, and 5. ABTS (2, 2’-azino-bis-3-ethylbenzothiazoline-6-sulfonic acid)assay was conducted to measure radical scavenging capacity (RSC) of the complex at pH 2, 4, 6, and 8, after which the RSCs were recorded over the 24 h heating(95°C) and 24 h UVB irradiating periods. The RSC values of the complexes increased as the β-CD concentration increased (0.1-2%) at pH 6 and 8. Upon thermal treatment, the RSC readings of the complexes tended to decrease to a lesser extent compared to the anthocyanin control (without β-CD) at pH 2. This result suggests that the thermal stability of anthocyanin was more retained in the presence of β-CD at low pH (pH 2). However, in the case of UVB irradiation, the effect of β-CD complex on the anthocyanin RSC was not significant, though the RSC values for both the anthocyanin control and complexes trended downward as the UVB irradiation time increased at all pH ranges except for pH 8 (no downward trend). In short, this study suggests that the effect of β-CD complex on the stability of anthocyanin antioxidant capacity depends on pH and the susceptibility to the degradation process.

The use of bee venom (Apis mellifera L., BV) occasionally causes side effects such as inflammation and allergic reactions in the recipients. Several case reports also suggested the treatment of BV has some limitations in its clinical uses, due to the occurrence of dermal necrosis and anaphylatic reactions. It is generally understood that bee venom allergy is mainly the result of its allergic component, phospholipase A2 (PLA2). The present study was aimed to generate PLA2-free bee venom (PBV) and evaluate its efficacy as skin care and cosmetic preparation, comparing with original bee venom (BV). Our results showed that both BV and PBV exhibited significant protective effects in UVB-irradiated human keratinocyte (HaCaT) and human dermal fibroblast (HDF) cells and they also induced type I collagen synthesis in UVB-irradiated HDF cells except BV at 3 μg/ml. Furthermore, BV and PBV showed the inhibition of UVB-stimulated matrix metalloproteinase-1 (MMP-1), a major collagen degrading enzyme in skin. However, BV, unlike PBV, exhibited strong cytotoxicities in skin cells (both HaCaT and HDF) at its working concentrations of anti-wrinkle effect. The underlying cell signaling mechanisms of anti-wrinkle effects of BV and PBV were demonstrated by the activation of ERK1/2, and p38. Conclusively, PBV appears to be the bee venom of choice with less cytotoxicity and higher efficacy on UVB-irradiated skin cells in comparison with original bee venom (BV). Therefore, PBV can better be used as a cosmetic ingredient exhibiting excellent anti-wrinkle effect against photoaging than original BV.

Wrinkles are an outward sign of cutaneous aging appearing preferentially on ultraviolet B (UVB)-exposed areas. The anti-wrinkle effects of herbal extracts were investigated in an animal model. Female albino hairless mice (HR/ICR) were randomly allocated to the control group (non-irradiated vehicle), positive control group (UVB irradiated-vehicle), and two herbal extract mixture groups (HE-1 and HE-2). HE-1 included Glycyrrhizae radix, Rhei Rhizoma, Cornus officinalis, and Sesami semeni, and HE-2 included Swertia pseudo-chinensis, Sophora flavescens, Scutellaria baicalensis, and Salvia miltiorrhiza. The herbal extract mixtures were pre-treated dorsally with 0.2 ml per individual five times per week for four weeks prior to the start of UVB irradiation. At the fifth week, the animals were exposed to UVB irradiation for a subsequent eight weeks, three times per week. The intensity of irradiation showed a gradual increase, from 30 mJ/cm 2 to 240 mJ/cm2 (1 MED: 60 mJ/cm2 ). Dorsal skin samples were stained with H&E in order to examine the epidermal thickness. In addition, Masson-Trichrome staining was performed for determination of the amount of collagen fiber. Treatments with HE-1&2 resulted in an increase in the amount of collagen fiber, a better appearance, and fewer wrinkles, compared with the positive control. As determined by hydroxyproline assay, treatments with HE-1&2 led to a significant increase in the amount of collagen, compared with the positive control group (p<0.05). Chronic UVB irradiation to skin of hairless mice resulted in an increase in expression of matrix metalloproteinase-1 (MMP-1), however, treatments with HE-1&2 tended to decrease the expression of MMP-1. These results indicate that the herbal extracts used in this study have a preventive effect on UVB-induced wrinkle formation in a hairless mouse model, due in part to inhibition of MMP-1 expression and increment of collagen amount.

Anti-wrinkle effect of herbal extracts was investigated on the skin of in a model of animal irradiated by ultraviolet rey B (UVB). The female albino hairless mice (HR/ICR) were randomly allocated to the normal control group (NC-non irradiated-vechicle), positive control group (PC, UVB irradiated vehicle) and herbal extract (HE) group. The herbal extract included the mixture of water and alcohol extract from Pleuropterus multflorus, Lonicera japonica Thunbert, Phellinus linteus, and Phaseolus radiatus. The herbal extract was treated dorsally with 0.2 ml per mouse five times a week for 12 weeks. At fifth week of the treatment, the animals were exposed to UVB irradiation for subsequent eight weeks three times a week. The intensity of irradiation was gradually increased from 30 mJ/㎠ to 240 mJ/㎠ (1MED: 60 mJ/㎠). Dorsal skins were obtained and stained with H&E to examine histological changes and epidermal/dermal thckness. The collagen fiber was also observed in Masson-Trichrome staining. Hydroxyproline assay and western blot analysis were also carried out to detect the change of collagen amount and to investigate MMP-1 expression, respectively. The HE group showed a better appearance and weak wrinkling, compared to PC group, The treatment of herbal extract significantly increased the thickness of dermis and the amount of collagen fibers compared to PC group (p＜0.05). The treatment of HE significantly increased the hydroxyproline amount compared to PC group (p＜0.05). The chronic UVB irradiation to hairless mice skin increased expression of MMP-1 but the treatment of HE decreased the expression of MMP-1. These results indicate that the herbal extract used in this study have a preventive effect on the UVB-induced wrinkle in a hairless mice model, partly due to the reduction of MMP-1 expression and increment of collagen amount.