Don-Gil Lee

The protective effects of fucosterol against skin damage in UVB-irradiated human dermal fibroblasts.

Exposure to ultraviolet (UV) light causes matrix metalloproteinase (MMP) overexpression and extracellular matrix depletion, leading to skin photoaging. The activation of MMP is related to increased interlukin-6 (IL-6) and type I procollagen production, which is regulated by transforming growth factor-β1 (TGF-β1). Activator protein-1 (AP-1) activation induces MMP-1 production and reduces type I procollagen secretion. Fucosterol, which is extracted and purified from the brown algae Hizikia fusiformis, is a phytosterol. We assessed the effects of fucosterol on photodamage and investigated its molecular mechanism of action in UVB-irradiated normal human dermal fibroblasts by using enzyme-linked immunosorbent assay, Western blot analysis, and reverse transcription-polymerase chain reaction. Our results showed that fucosterol significantly decreased the UVB-induced expression of MMP-1, IL-6, p-c-Jun, and p-c-Fos. Additionally, fucosterol markedly increased the UVB-induced production of type I procollagen and TGF-β1. Our results indicate that fucosterol regulates MMP-1 and type I procollagen expression by modulating AP-1 and TGF-β1 signaling and that MMP-1 activation is correlated with IL-6. These data suggest that fucosterol is a promising botanical agent to protect against skin photodamage.

Effects of Galla chinensis extracts on UVB-irradiated MMP-1 production in hairless mice

Galla chinensis (GAC) is a natural traditional Chinese medicine that has been widely used in folk medicine. Although GAC compounds (mainly gallic acid and methyl gallate) possess strong antiviral, antibacterial, anticancer, and antioxidant activities, there is no report regarding topical or oral administration of GAC compounds on UVB irradiation-induced photoaging in hairless mice (SKH: HR-1). In the present study, we examined cell viability, intracellular reactive oxygen species (ROS), matrix metalloproteinase-1 (MMP-1), and interleukin-6 (IL-6) in skin fibroblasts and keratinocytes induced by UVB in vitro. We also studied skin damage by measuring skin thickness, elasticity, wrinkling and levels of protein MMP-1, elastin, procollagen type I, and transforming growth factor-β1 (TGF-β1) in hairless mouse skin chronically irradiated by UVB in vivo. GAC treatment significantly prevented skin photoaging by reducing the levels of ROS, MMP-1, and IL-6 and promoting production of elastin, procollagen type I, and TGF-β1. According to the results of H&E staining and Masson’s trichrome staining, GAC reduced skin thickness and wrinkle formation while it increased skin elasticity. The effects of GAC on UVB-induced skin photoaging may be due to suppressed MMP-1 expression. These findings could be referenced for the development of new agents that target UVB-induced photoaging.

Enzyme-processed Korean Red Ginseng extracts protects against skin damage induced by UVB irradiation in hairless mice

UV irradiation is the main factor contributing to skin damages that are associated with an excessive production of matrix-degrading metalloproteinase (MMP)-1 and a deficient expression of collagens. To date, red ginseng has been revealed to possess many biomedical effects, such as anti-aging, anti-oxidation, and anti-inflammatory. In this study, we prepared the Korean Red Ginseng extracts treated with enzyme (KRGE) and investigated the effects of dietary KRGE on the formation of wrinkles generated by UVB irradiation in hairless mice. It was found that KRGE inhibited the UVB-induced formation of wrinkles, epidermal thickness, and skin dryness in hairless mice. Further results also showed that KRGE attenuated UVB-induced MMP-1 level, while accelerated procollagen type I, transforming growth factor-β1 secretion. Interestingly, the expression of profilaggrin and filaggrin in both the epidermis and dermis were decreased due to UVB exposure and reversed by KRGE. The KRGE 0.06% was prior to KRGE 0.24%. In view of these results, which indicated that KRGE protected skin from UVB-induced photodamages, which may not only mediated by regulating of MMP-1 and procollagen type I, but also by increasing the production of profilaggrin and filaggrin. In conclusion, our results suggest that KRGE may be a promising agent for the treatment of skin photodamages. The challenge of KRGE will be expected as cosmeceuticals and nutraceuticals in order to intervene in aging-related degenerative skin changes.

The inductive effect of ginsenoside F2 on hair growth by altering the WNT signal pathway in telogen mouse skin

This study was conducted to confirm the possibility of using minor ginseng saponin F2 by oral administration on hair anagen induction effects. The signaling pathway and anagen induction effect of ginsenoside F2 were investigated and compared with finasteride on the effect of hair growth induction. The cell-based MTT assay results indicated that the proliferation rates of HHDPC and HaCaT treated with F2 significantly increased by 30% compared with the finasteride-treated group. A western blot study showed that the expression of β-catenin Lef-1 and DKK-1 increased by 140, 200% and decreased by 40% in the F2-treated group, respectively compared to that of finasteride-treated group. C57BL/6 mice were subjected to the same treatments. The hair growth promotion rates were compared with groups treated with finasteride, which was 20% higher in the F2-treated group. Tissue histological analysis results showed the number of hair follicles, thickness of the epidermis, and follicles of the anagen phase which increased in the F2-treated group, compared with the finasteride-treated groups. Moreover, the effect of F2 on hair growth was confirmed through the immunofluorescence (IF) methods indicating the expression aspect of Wnt signal pathway-related factors in the tissue of C57BL/6 mouse. Our results considered the expression increase in β-catenin, Lef-1 which was suggested as a major factor related to the development and growth of hair follicle and the decrease in DKK-1 when entering catagen by F2. As the data showed, F2 might be a potential new therapeutic source for anagen induction and hair growth through the Wnt signal pathway.

Pedobacter ginsenosidimutans sp. nov., with ginsenoside-converting activity.

A Gram-reaction-negative, strictly aerobic, non-motile, non-spore-forming and rod-shaped bacterial strain, designated THG-45(T), was isolated from soil of a ginseng field of Pocheon province in the Republic of Korea and its taxonomic position was investigated by a polyphasic approach. Growth occurred at 4-30 °C, at pH 5.5-9.0 and with 0-2 % (w/v) NaCl on nutrient agar. On the basis of 16S rRNA gene sequence similarity, strain THG-45(T) was shown to belong to the genus Pedobacter and was related to Pedobacter borealis G-1(T) (98.8 %), P. alluvionis NWER-II11(T) (97.9 %), P. agri PB92(T) (97.9 %), P. terrae DS-57(T) (97.5 %), P. suwonensis 15-52(T) (97.4 %), P. sandarakinus DS-27(T) (97.0 %) and P. soli 15-51(T) (97.0 %), but DNA relatedness between strain THG-45(T) and these strains was below 36 %. The G+C content of the genomic DNA was 39 mol%. The only isoprenoid quinone detected in strain THG-45(T) was menaquinone-7 (MK-7). The predominant fatty acids were iso-C15 : 0, summed feature 3 (comprising C16 : 1ω6c and/or C16 : 1ω7c) and iso-C17 : 0 3-OH, and the major polar lipids were phosphatidylethanolamine and an unidentified aminophosphoglycolipid. Phenotypic data and phylogenetic inference supported the affiliation of strain THG-45(T) to the genus Pedobacter, and a number of biochemical tests differentiated strain THG-45(T) from the recognized species of the genus Pedobacter. Therefore, the novel isolate represents a novel species, for which the name Pedobacter ginsenosidimutans sp. nov. is proposed, with THG-45(T) as the type strain ( = KACC 14530(T) = JCM 16721(T)).

The Androgenic Alopecia Protective Effects of Forsythiaside‐A and the Molecular Regulation in a Mouse Model

This study examined the inhibitory effect of forsythiaside‐A, a natural substance derived from Forsythia suspensa (F. suspensa), on entry into catagen induced by dihydrotestosterone (DHT) in an androgenic alopecia mouse model. In vitro experiment comparing finasteride with forsythiaside‐A showed that forsythiaside‐A treatment resulted in a 30% greater inhibition of DHT‐induced apoptosis in human hair dermal papilla cell (HHDPCs) and human keratinocytes (HaCaTs). In vivo experiment showed that mouse hair density and thickness were increased by 50% and 30%, respectively, in the forsythiaside‐A‐treated group when compared to a DHT group. Tissue histological results revealed that the forsythiaside‐A‐treated group had an increase in size and shape of the hair follicles and a 1.5 times increase in the follicle anagen/telogen ratio when compared to the finasteride group. Western blot examination of TGF‐β2 expression related to apoptosis signaling in mouse skin verified that forsythiaside‐A reduced the expression of TGF‐β2 by 75% and suppressed apoptosis by reducing the expression of caspase‐9 by 40%, and caspase‐3 by 53%, which play an roles up‐regulator in the apoptosis signal. The forsythiaside‐A group also showed a 60% increase in the Bcl‐2/Bax ratio, which is a factor related to mitochondrial apoptosis. Our results indicated that forsythiaside‐A prevents apoptosis by similar mechanism with finasteride, but forsythiaside‐A is more effective than finasteride. In summary, forsythiaside‐A controlled the apoptosis of hair cells and retarded the entry into the catagen phase and therefore represents a natural product with much potential for use as a treatment for androgenic alopecia. Copyright

The bone regenerative effects of fucosterol in in vitro and in vivo models of postmenopausal osteoporosis

SCOPE The aim of this study was to investigate the bone regenerative effects of fucosterol in estrogen-deficient ovariectomized (OVX) rats. METHODS AND RESULTS Bone regeneration was assessed in fucosterol-treated MG63 cells in vitro via assays for osteoblast proliferation, alkaline phosphatase, and osteoclast differentiation. Osteoblast proliferation rates, alkaline phosphatase activity, and mineralization were increased in the fucosterol-treated group. Moreover, differentiation of osteoclasts was decreased in the fucosterol-treated group. In the in vivo assay, female rats were OVX. Twelve weeks after ovariectomy, rats were divided into seven groups, each oral administrate everyday for 7 weeks. The bone mineral density of femoral bones was higher in fucosterol groups than in OVX control, and body weight was lower in fucosterol groups. Among bone-quality parameters, bone volume/total volume increased and trabecular separation decreased in fucosterol groups relative to the OVX control. Bone formation and resorption were evaluated using the serum biomarkers osteocalcin and CTx. Fucosterol tripled the level of serum osteocalcin relative to the OVX group and reduced the serum level of CTx. CONCLUSION These results suggest that fucosterol has the dual potentials to activate osteoblasts to stimulate bone formation and suppress differentiation of osteoclasts so as to reduce bone resorption.

Effect of oral administration of fucosterol from Hizikia fusiformis on DNCB-induced atopic dermatitis in NC/Nga mice

Fucosterol was extracted and purified from the brown algae Hizikia fusiformis. We assessed the anti-inflammatory effects of fucosterol on 2,4-dinitrochlorobenzene (DNCB)-induced atopic dermatitis (AD)-like lesions in NC/Nga mice. Fucosterol decreased the lipopolysaccharide-induced production of nitric oxide in mouse macrophage cells. Oral administration of fucosterol (200 mg/kg weight/day) decreased the frequency of scratching, epidermal thickness of the dorsal skin, and number of degranulated mast cells. In addition, fucosterol treatment decreased levels of DNCB-stimulated serum immunoglobulin E (IgE). Compared to the control concanavalin A, fucosterol inhibited levels of IL-4 and TNF-α and increased secretion of IFN-γ, in the media of cultured splenocytes. Taken together, our results suggest that fucosterol may attenuate AD-like lesions by exerting anti-inflammatory effects, including inhibition of IgE and inflammatory cytokines, which modulate the Th1/Th2 balance.

Fucosterols from Hizikia fusiformis and their proliferation activities on osteosarcoma-derived cell MG63

Four fucosterol derivatives were isolated from the ethyl acetate fraction of Hizikia fusiformis. The chemical structures of the sterols were elucidated as fucosterol (1), a mixture of 24R,28R- and 24S,28R-epoxy-24-ethylcholesterol at the ratio of 3 to 2 (2), and 24R-saringosterol (3), all of which exhibited proliferation activity on MG63 cells.

Immunostimulatory Effect of Enzyme-Modified Hizikia fusiformein a Mouse Model In Vitro and Ex Vivo

Hizikia fusiforme, a brown seaweed, has been utilized as a health food and in traditional medicine. In this study, we investigated whether enzyme-modified H. fusiforme extracts (EH) have immunological effects compared with normal H. fusiforme extracts (NH). The effects of NH and EH on immune responses were investigated by assessing nitric oxide (NO) production, phagocytosis, and cytokine secretion in RAW 264.7 murine macrophages and mice. Also, fucosterol was evaluated to find the active component of NH and EH by addressing cytotoxicity test and NO production. Both of NH and EH significantly increased cell viability and NO synthesis. Tumor necrosis factor-α (TNF-α) expression was more induced by EH with LPS treatment. Phagocytic activity, as the primary function of macrophages, was markedly induced by EH treatment. Additionally, EH encouraged splenocyte proliferation and recovered the levels of cytokines IL-1β, IL-6, and TNF-α in mice. Finally, fucosterol increased NO production with no cytotoxicity, which means that fucosterol is an active component of EH. In conclusion, EH has the potential to modulate immune function and could offer positive therapeutic effect for immune system diseases.