• Proceedings of the Korean Society of Applied Pharmacology
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• 2005.04a
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• pp.131-148
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• 2005

This study is peformed to investigate the effect of water extract from Vaccinium uliginosum L., on melanin production in B 16 melanoma cells, procollagen production and matrix metalloproteinase-1(MMP-1) inhibition in human fibroblast cells. One hundred grams of the Vaccinium uliginosum L. was extracted with 2000 mL of water($90^{\circ}C$, 16h, 2times). The water extracts were lyophilized and stored at $4^{\circ}C$ until used. Dry weight yields of extracts of Vaccinium uliginosum L. were $3\%$(w/w). Extracts from Vaccinium uliginosum L. showed scavenger activities on DPPH radical, superoxide anion radical, hydroxyl radical, hydrogen peroxide and singlet oxygen radical. And these substances inhibited release of cyiokines from human keratinocyte after UV B exposure. Therefore we confirmed that extracts from Vaccinium uliginosum L. had antioxidative effect. These substances inhibited purified tyrosinase activity and melanogenesis in B 16 melanoma cells treated/untreated IL-$1{\alpha}$. Moreover this extract stimulated procollagen production and inhibited MMP-1 production in human fibroblast cells treated/untreated IL-$1{\beta}$. Therefore we confirmed that extracts from Vaccinium uliginosum L. had whitening effect. And these substances decreased degree of wrinkle in hairless mouse skin that induced by UV B irradiation. Therefore we confirmed that extracts from Vaccinium uliginosum L. had anti-wrinkle effect. From the above results, it is possible that Vaccinium uliginosum L. may be developed to be an anti-melanogenesis agent and anti-wrinkle agent.

• The Journal of Korean Medicine
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• v.24 no.2
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• pp.159-165
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• 2003

Objectives : Coptidis rhizoma (CR) is an oriental medicine that has been used in many traditional prescriptions against diabetes mellitus in Korea for centuries. Our purpose was to determine the protective effect and its action mechanism of CR on the cytotoxicity of pancreatic -cell line (RINm5F cell). Methods : In this experiment, we used methods such as MTT assay for detection of cytotoxicity, DNA fragmentation assay for detection of apoptotic cell death, LDH activity assay for detection of necrotic cell death, and measurement of $DiOC_{6}$ (3) retention for detection of mitochondrial membrane potential (MMP). Background : Nitric oxide (NO) is believed to playa key role in the process of pancreatic -cell destruction leading to insulin-dependent diabetes mellitus (IDDM). Results : Exposure of RINm5F cells to chemical NO donor such as S-nitroso-N-acetylpenicillamine (SNAP) induced cytotoxic events such as DNA fragmentation and lactate dehydrogenase (LDH) release into medium. However, pretreatment of RINm5F cells with CR extract ($10~50{\mu\textrm{g}}/ml$) for 3 hours prevented SNAP-induced DNA fragmentation and LDH release into medium through the inhibition of MMP disruption. Conclusions : These results suggest that CR may be a candidate for a therapeutic or preventing agent against IDDM.

• BMB Reports
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• v.51 no.11
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• pp.572-577
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• 2018

Recent studies showed that the PD-1/PD-L1 checkpoint blockade is a dramatic therapy for melanoma by enhancing antitumor immune activity. Currently, major strategies for the PD-1/PD-L1 blockade have mainly focused on the use of antibodies and compounds. Seeking an alternative approach, others employ endogenous proteins as blocking agents. The extracellular domain of PD-1 (ePD1) includes the binding site with PD-L1. Accordingly, we constructed a PD-1-based recombinantly tailored fusion protein (dFv-ePD1) that consists of bivalent variable fragments (dFv) of an MMP-2/9-targeted antibody and ePD1. The melanoma-binding intensity and antitumor activity were also investigated. We found the intense and selective binding capability of the protein dFv-ePD1 to human melanoma specimens was confirmed by a tissue microarray. In addition, dFv-ePD1 significantly suppressed the migration and invasion of mouse melanoma B16-F1 cells, and displayed cytotoxicity to cancer cells in vitro. Notably, dFv-ePD1 significantly inhibited the growth of mouse melanoma B16-F1 tumor cells in mice and in vivo fluorescence imaging showed that dFv-ePD was gradually accumulated into the B16-F1 tumor. Also the B16-F1 tumor fluorescence intensity at the tumor site was stronger than that of dFv. This study indicates that the recombinant protein dFv-ePD1 has an intensive melanoma-binding capability and exerts potent therapeutic efficacy against melanoma. The novel format of the PD-L1-blocked agent may play an active role in antitumor immunotherapy.

• Proceedings of the Plant Resources Society of Korea Conference
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• 2019.10a
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• pp.67-67
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• 2019

Vaccinium oldhamii (V. oldhamii) has been reported to exert a variety of the pharmacological properties such as anti-oxidant activity, anti-cancer activity, and inhibitory activity of ${\alpha}$-amylase and acetylcholinesterase. However, the anti-inflammatory activity of V. oldhamii has not been studied. In this study, we aimed to investigate anti-inflammatory activity of the stem extracts from V. oldhamii, and to elucidate the potential mechanisms in LPS-stimulated RAW264.7 cells. Among VOS, VOL and VOF, the inhibitory effect of NO and PGE2 production induced by LPS was highest in VOS treatment. Thus, VOS was selected for the further study. VOS dose-dependently blocked LPS-induced NO and PGE2 production by inhibiting iNOS and COX-2 expression, respectively. VOS inhibited the expression of pro-inflammatory cytokines such as $IL-1{\beta}$, IL-6 and $TNF-{\alpha}$. In addition, VOS suppressed TRAP activity and attenuated the expression of the osteoclast-specific genes such as NFATc1, c-FOS, TRAP, MMP-9, cathepsin K, CA2, OSCAR and ATPv06d2. VOS inhibited LPS-induced $NF-{\kappa}B$ signaling activation through blocking $I{\kappa}B-{\alpha}$ degradation and p65 nuclear accumulation. VOS inhibited MAPK signaling activation by attenuating the phosphorylation of ERK1/2, p38 and JNK. Furthermore, VOS inhibited ATF2 phosphorylation and blocked ATF2 nuclear accumulation. From these findings, VOS has potential to be a candidate for the development of chemopreventive or therapeutic agents for the inflammatory diseases.

• Proceedings of the SCSK Conference
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• 2003.09b
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• pp.519-523
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• 2003

Alpha hydroxy acid (AHA) includes a group of organic acids found in natural foods such as sugarcane (glycolic acid), milk (lactic acid), apples (malic acid) and oranges (citric acid). Earlier studies demonstrated the effect of AHAs on the skin by diminishing the adhesiveness of the corneal layer and increasing the viable epidermal thickness. Recent data suggest that AHAs have some effects on the dermal component of skin and even affect the aging process of the skin. A previous study revealed increased collagen production by treatment with glycolic acid among AHAs in vitro. However, the mechanism of the regulation of collagen production by glycolic acid was unclear. In present study, we tried to demonstrate the effect of glycolic acid on human dermal fibroblasts and to unveil the mechanism of regulation of collagen production by glycolic acid in human dermal fibroblasts: proliferation of fibroblasts and collagen synthesis and degradation by collagenases in fibroblasts. Our results suggested that glycolic acid had no effect on proliferation and cytotoxicity of adult human dermal fibroblasts. However, glycolic acid not only induced the increase of the collagen synthesis in human dermal fibroblasts at lower concentration than 0.1 % but also inhibited MMP-2 activity of human dermal fibroblast in the range between 0.01 and 0.4% and MMP-9 activity of human dermal fibroblast in the range between 0.06 and 0.09%. In summary, our results suggest that glycolic acid may increase wrinkle reduction partially by both increase in collagen synthesis and decrease in collagen degradation.

• International Journal of Oral Biology
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• v.39 no.3
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• pp.159-167
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• 2014

Curcumin is a widely used flavoring agent in food, and it has been reported to inhibit cell growth, to induce apoptosis, and to have antitumor activity in many cancers. Cisplatin is one of the most potent known anticancer agents and shows significant clinical activity against a variety of solid tumors. This study was undertaken to investigate the synergistic apoptotic effects of co-treatment with curcumin and cisplatin on human tongue SCC25 cells. To investigate whether the co-treatment efficiently reduced the viability of the SCC25 cells compared with the two treatments separately, an MTT assay was conducted. The induction and the augmentation of apoptosis were confirmed by DNA electrophoresis, Hoechst staining, and an analysis of DNA hypoploidy. Western blot, MMP and immunofluorescence tests were also performed to evaluate the expression levels and the translocation of apoptosis-related proteins following the co-treatment. In this study, following the co-treatment with curcumin and cisplatin, the SCC25 cells showed several forms of apoptotic manifestation, such as nuclear condensation, DNA fragmentation, reduction of MMP, increased levels of Bax, decreased levels of Bcl-2, and decreased DNA content. In addition, they showed a release of cytochrome c into the cytosol, translocation of AIF and DFF40 (CAD) to the nuclei, and activation of caspase-7, caspase-3, PARP, and DFF45 (ICAD). In contrast, separate treatments of $5{\mu}M$ of curcumin or $4{\mu}g/ml$ of cisplatin, for 24 hours, did not induce apoptosis. Therefore, our data suggest that combination therapy with curcumin and cisplatin could be considered as a novel therapeutic strategy for human oral squamous cell carcinoma.

• Environmental Analysis Health and Toxicology
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• v.24 no.2
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• pp.169-178
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• 2009

To investigate the inhibitory effects of Peonia japonica water extract(PJWE) on skin wrinkle formation, skin wrinkles were induced by both the irradiation of UVB and the application of squalene monohydroperoxide to the backs of hairless mice for 4 weeks. And at the same time experimental materials were applied topically. Wrinkles for the control (C) group were formed as a pattern of deep furrows and thick crests. Whereas wrinkles for the positive control (PC, 0.01% retinoic acid) and experimental(E, PJWE) groups were formed as a pattern of shallow furrows and thin crests, which were similar to that of the normal(N) group. Collagen and elastic fibers in dermis of the PC and E groups were almost intact with a regular arrangement, which were similar to those of the N group. The activity of xanthine oxidase, the free radical generating enzyme, was significantly lower in the E group than the C and PC groups. The activities of superoxide dismutase and catalase, the free radical scavenging enzymes, were much higher in the E group than the C and PC groups and similar to the N group. As for the amount of matrix metalloproteinase-3(MMP-3) expression, PC and E groups were significantly lower than the C group. Therefore, PJWE could be very effective natural herbal material for the inhibition or improvement of wrinkle formation in hairless mice skin.

• International Journal of Oral Biology
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• v.38 no.3
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• pp.101-110
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• 2013

We investigated the synergistic apoptotic effects of co-treatments with Chios gum mastic (CGM) and eugenol on G361 human melanoma cells. An MTT assay was conducted to investigate whether this co-treatment efficiently reduces the viability of G361 cells compared with each single treatment. The induction and augmentation of apoptosis were confirmed by DNA electrophoresis, Hoechst staining, and analyses of DNA hypoploidy. Western blot analysis and immunofluorescent staining were also performed to evaluate expression and translocation of apoptosis-related proteins following CGM and eugenol co-treatment. Proteasome activity and mitochondrial membrane potential (MMP) changes were also assayed.The results indicated that the co-treatment of CGM and eugenol induces multiple pathways and processes associated with an apoptotic response in G361 cells. These include nuclear condensation, DNA fragmentation, a reduction in MMP and proteasome activity, an increase of Bax and decrease of Bcl-2, a decreased DNA content, cytochrome c release into the cytosol, the translocation of AIF and DFF40 (CAD) into the nucleus, and the activation of caspase-9, caspase-7, caspase-3, PARP and DFF45 (ICAD). In contrast, separate treatments of $40{\mu}g/ml$ CGM or $300{\mu}M$ eugenol for 24 hours did not induce apoptosis. Our present data thus suggest that a combination therapy of CGM and eugenol is a potential treatment strategy for human melanoma.

• Proceedings of the Korean Society of Applied Pharmacology
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• 2007.11a
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• pp.153-158
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• 2007

Background: Naturally occurring antioxidants were used to regulate the skin damage caused by ultraviolet (UV) radiation because several antioxidants have demonstrated that they can inhibit wrinkle formation through prevention of matrix metalloproteinases (MMPs) and/or increase of collagen synthesis. We examined the effect of oral administration of the antioxidant mixture ("L-Skin Care") on UVB-induced wrinkle formation. In addition, we investigated the possible molecular mechanisms of photoprotection against UVB through inhibition of collagen-degrading MMP activity or through enhancing of pro collagen synthesis in mouse dorsal skin. Methods: Female SKH-l hairless mice were orally administrated "L-Skin Care" (test group) or vehicle (control group) for 10 weeks with UVB irradiation by three times a week. The intensity of irradiation was gradually increased from 30 to $180mJ/cm^2$. Microtopographic and histological assessments of the dorsal skins were carried out at the end of 10 weeks to evaluate wrinkle formation. Western blot analysis and EMSA were also carried out to investigate the changes in the balance of collagen synthesis and collagen degradation. Results: Our "L-Skin Care" significantly reduced UVB-induced wrinkle formation, accompanied by significant reduction of epidermal thickness, and UVB-induced hyperplasia, acanthosis and hyperkeratosis. Oral administration of "L-Skin Care" significantly prevented UVB-induced expressions of MMPs, mitogen-activated protein (MAP) kinases and activation of activator protein (AP)-1 transcriptional factor in addition to enhanced type I procollagen and transforming growth factor-$\beta$ (TGF-$\beta$) expression. Conclusion: Oral administration of "L-Skin Care" significantly inhibited wrinkle formation caused by chronic UVB irradiation through significant inhibition of UVB-induced MMP activity accompanied with enhancement of collagen synthesis.

• Journal of the Korean Society of Food Science and Nutrition
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• v.45 no.12
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• pp.1708-1716
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• 2016

The Akt/mammalian target of the rapamycin (mTOR) pathway is activated in the majority of human cancers. Activation of the Akt/mTOR pathway confers resistance to many types of cancer therapy. In this study, we evaluated the apoptotic effect of ethanol extract of Artemisia annua L. through down-regulation of Akt signal pathways and the mitochondrial pathway in hepato-carcinoma cells (HepG2). A. annua extract is known as a medicinal herb that is effective against cancer. We evaluated anti-proliferative activity by MTT-based viability assay and apoptotic effect by Annexin-V/PI staining, mitochondrial membrane potential (MMP), and caspase-3/7 activity as determined by flow cytometry. A. annua treatment led to loss of MMP, resulting in cytochrome c-inducible activation of caspase-3/7. Treatment with A. annua extract reduced activities of Akt/mTOR/anti-apoptotic proteins (such as Bcl-2 and $Bcl-X_L$), leading to increased activation of tumor suppressor p53 and pro-apoptotic proteins (such as Bax and Bak). We applied LY294002 (inhibitor of Akt) and rapamycin (inhibitor of mTOR) to determine the relationship between signal transduction of proteins associated with apoptosis. LY294002 and rapamycin significantly reduced cell viability and increased apoptosis. These results indicate that Bcl-2 and caspase-3 are key regulators in A. annua extract-induced apoptosis in HepG2 cells and are controlled through the Akt/mTOR signaling pathway.