• Journal of Microbiology and Biotechnology
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• 제30권3호
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• pp.325-332
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• 2020

Methyl linderone (ML), a cyclo-pentenedione, was isolated from the fruit of Lindera erythrocarpa Makino (family Lauraceae). This plant has well-known anti-inflammatory effects; however, the anti-cancer effects of ML have not yet been reported. Thus, in the present study we investigated the effects of ML on the metastasis of human breast cancer cells. We used 12-O-tetradecanoyl phorbol-13-acetate (TPA)-stimulated MCF-7 cells as the cell model to study the effects of ML on invasion and migration. ML was found to reduce the invasion and migration rate of TPA-stimulated MCF-7 cells. Moreover, it inhibited two metastasis-related factors, matrix metalloproteinase-9 (MMP-9) and interleukin-8 (IL-8), at the mRNA and protein expression levels, in TPA-treated MCF-7 cells. The mechanism by which ML exerted these effects was through the inhibition of translocation of activator protein-1 (AP-1) and signal transducer and activator of transcription-3 (STAT3), mediated via phosphorylation of extracellular signal-regulated kinase (ERK). Taken together, our findings indicated that ML attenuated the TPA-stimulated invasion and migration of MCF-7 cells by suppressing the phosphorylation of ERK and its downstream factors, AP-1 and STAT3. Therefore, ML is a potential agent for the treatment of breast cancer metastasis.

• Microbiology and Biotechnology Letters
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• 제42권2호
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• pp.194-201
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• 2014

Clitocybin A is a novel anti-wrinkle cosmetic agent produced by the strain from a Korean native mushroom Clitocybe aurantiaca. In this study, fermentation, extraction, and purification conditions for a large scale production of clitocybin A were optimized, and its cytotoxicity and inhibition activity on the expression of matrix metalloproteinase-1 (MMP-1) were characterized. The mass production of anti-wrinkle agent was achieved according to the 300 L fermentation process with a fed-batch cultivation using the modified yeast-maltose (YM) broth, and a total of 12.5 kg of cell mass was obtained in a 120 L culture broth for 14 days. After extraction and purification, clitocybin A was identified by HPLC. The cytotoxicity of clitocybin A was examined by the MTT assay. When assayed at 100 and 200 ${\mu}g/ml$ concentrations, clitocybin A showed no cytotoxicity, demonstrating safety. The inhibition activity of clitocybin A on the expression of MMP-1 was examined against UV irradiation. Oleanolic acid (control group) showed a relatively low MMP-1 inhibiting activity (ca. 16.7%) at 10 ${\mu}g/ml$ and showed increased cytotoxicity at higher concentrations. In contrast, clitocybin A showed no cytotoxicity at 100 ${\mu}g/ml$, and exhibited a relatively high MMP-1-inhibiting activity (33.1%). These findings indicate that clitocybin A may be a safe and effective anti-wrinkle agent for use in functional cosmetics.

• The Korean Journal of Oral and Maxillofacial Pathology
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• 제41권3호
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• pp.121-129
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• 2017

Coffee (Coffea spp.) is one of the most important agricultural commodities, being widely consumed in the world. Various beneficial health effects of coffee have been extensively investigated, but data on habitual coffee consumption and its bio-physiological effect have not been clearly explained as well as it is not proved the cause and effect between drinking coffee and its bio-physiological reactions. We made the dialyzed coffee extract (DCE), which is absorbable through gastrointestinal tract, in order to elucidate the cellular effect of whole small coffee molecules. RAW 264.7 cells, a murine macrophage lineage, were directly treated with DCE, i.e., DCE-2.5 (equivalent to 2.5 cups of coffee a day), DCE-5, and DCE-10, for 12 hours, and their protein extracts were examined by immunoprecipitation high performance liquid chromatography (IP-HPLC). RAW 264.7 cells differently expressed the inflammation-related proteins depending on the doses of DCE. RAW 264.7 cells treated with DCE showed marked increase of cathepsin C, cathepsin G, CD20, CD28, CD31, CD68, indicating the activation of innate immunity. Particularly, the macrophage biomarkers, cathepsin G, cathepsin C, CD31, and CD68 were markedly increased after DCE-5 and DCE-10 treatments, and the lymphocyte biomarkers, CD20 and CD28 were consistently increased and became marked after DCE-10 treatment. On the other hand, RAW 264.7 cells treated with DCE showed consistent increase of IL-10, an anti-inflammatory factor, but gradual decreases of different pro-inflammatory proteins including $TNF{\alpha}$, COX-2, lysozyme, MMP-2, and MMP-3. In particular, the cellular signaling of inflammation was gradually mitigated by the reduction of $TNF{\alpha}$, COX-2, IL-12, and M-CSF, and also the matrix inflammatory reaction was reduced by marked deceases of MMP-2, MMP-3, and lysozyme. These anti-inflammatory expressions were consistently found until DCE-10 treatment. Therefore, it is presumed that DCE may have dynamic effects of innate immunity activation and pro-inflammation suppression on RAW264.7 cells simultaneously. These effects were consistently found in the highest dose of coffee, DCE-10 (equivalent to 10 cups of coffee a day in man), that might imply the small coffee molecules were accumulated in RAW 264.7 cells after DCE-10 treatment and produce synergistic cytokine effects for innate immunity activation and anti-inflammatory reaction concurrently.

• Nutrition Research and Practice
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• 제7권3호
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• pp.160-165
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• 2013

3T3-L1 preadipocyte were differentiated to adipocytes, and then treated with 0, 10, 20, and $40{\mu}g/mL$ of peanut sprout ethanol extract (PSEE). The main component of PSEE is resveratrol which contained 5.55 mg/mL of resveratrol. The MTT assay, Oil-Red O staining, glycerol-3-phosphate dehydrogenase (GPDH) activity, and the triglyceride concentration were determined in 3T3-L1 cells. MMP-2 and MMP-9 activities as well as mRNA expressions of C/EBP ${\beta}$ and C/EBP ${\alpha}$ were also investigated. As the concentration of PSEE in adipocytes increased, the cell proliferation was decreased in a dose-dependent manner from 4 days of incubation (P < 0.05). The GDPH activity (P < 0.05) and the triglyceride concentration (P < 0.05) were decreased as the PSEE treatment concentration increased. The mRNA expression of C/EBP${\beta}$ in 3T3-L1 cells was significantly low in groups of PSEE-treated, compared with control group (P < 0.05). The MMP-9 (P < 0.05) and MMP-2 (P < 0.05) activities were decreased in a dose-dependent manner as the PSEE concentration increased from $20{\mu}g/mL$. In conclusion, it was found that PSEE has an effect on restricting proliferation and differentiation of adipocytes.

• Journal of Oral Medicine and Pain
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• 제44권4호
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• pp.160-168
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• 2019

Purpose: To search the salivary factors that objectively indicate an pain in myalgia patients with temporomandibular joint disorder (TMD) and determine the possibility of the factors as pain-biomarkers. Methods: Participants consisted of pain-free 15 persons (male 7, female 8, mean age±standard deviation (SD); 26.8±16.04 years) and 45 myalgia patients with TMD (male 21, female 24, mean age±SD; 27.98±13.01 years). They were divided into a pain-free group (numerical rating scale [NRS] score 0), a mild pain group (NRS 1-4), a moderate pain group (NRS 5-6), and a severe pain group (NRS 7-10) and members of all groups were age, sex matched. Interleukin-8 (IL-8) and matrix metalloprotease 9 (MMP-9) were selected as pain biomarkers, by searching the Gene Expression Omnibus database and analyzing pain-related genes. Enzyme-linked immunosorbent assays were used to measure the concentration of IL-8 and MMP-9 in the patients' saliva. Results: IL-8 and MMP-9 levels were statistically significantly higher in pain groups than in the pain-free group. Greater differences were observed in patients with acute pain (with painful duration under 3 months) than in the control group and in female patients than in male. Conclusions: Salivary IL-8 and MMP-9 may play a role as biomarkers of myalgia in patients with TMD.

• The Korea Journal of Herbology
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• 제29권5호
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• pp.55-63
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• 2014

Objectives : The purpose of this study was verification of the restoration ability effect of Danggwisusan extract(DG) in wound induced Rat. Methods : It needed to make a scar(around $2{\times}2cm^2$) on the top of the fascia in the back of the rats and then the rats were divided into 4groups(n=6). Control was not treated at all, where as DG was orally medicated DG, Terra was percutaneously applied Terramycin, and DG + Terra was both orally medicated DG and percutaneously applied Terramycin per day for three weeks. Results : 93% or higher cell viability was observed in all tested groups from 1, 10, $100{\mu}g/m{\ell}$ in RAW 264.7cells. The DG decreased NO and cytokine production activity dose dependently. The production of IL-$1{\beta}$, IL-6 and TNF-${\alpha}$ were decreased by 46%, 46% and 40% in DG treated $100{\mu}g/m{\ell}$. The size of wound was significantly decreasing in DG, Terra, DG + Terra. WBC was significantly reduced in DG and DG + Terra. Monocyte was significantly reduced in DG, Terra and DG + Terra. Neutrophil was also reduced in DG, DG + Terra but not meaningless. The mRNA expression of MMP-1 was significantly reduced in Terra, MMP-2 was significantly reduced in DG, Terra, DG + Terra, and MMP-9 was significantly reduced in DG + Terra. Conclusions : According to the results, we thought that DG showed anti-inflammatory activities on the RAW 264.7cells in mouse macrophage and in adult rat wound. Moreover, the progress of recovery was found visually, heamatologically, genetically and histopathologically. In conclusion, it could be thought that DG has effect on the treating of wound.

• The Korea Journal of Herbology
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• 제37권6호
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• pp.37-44
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• 2022

Objectives : From this study, we sight to identify chondro-protective and anti-inflammatory effects of Sorbi Commixtae Fructus extract and its compound, chlorogenic acid. Methods : Sorbi Commixtae Fructus were extracted by 50% ethanol. And chlorogenic acid in Sorbi Commixtae Fructus 50% extract was quantified by high performance liquid chromatography (HPLC). To investigate chondro-protective effects, we treated Sorbi Commixtae Fructus 50% ethanol extract and chlorogenic acid in TNF𝛼-activated ATDC5 murine chondrogenic cells. After 24 hours, protein level of matrix metalloproteinase-3 (MMP3) and mRNA level of matrix metalloproteinase-13 (MMP13) were measured by using ELISA or reverse transcription PCR, respectively. To examine anti-inflammatory effects, we treated Sorbi Commixtae Fructus 50% ethanol extract and chlorogenic acid in LPS-induced RAW 264.7 murine macrophages. We measured the level of inflammatory mediators, such as Prostaglandin E2 (PGE2), Interleukin-6 (IL6) by ELISA and nitric oxide (NO) by Griess reagent assay. Results : A concentration of chlorogenic acid in Sorbi Commixtae Fructus 50% ethanol extract was 3.9 mg/g. Sorbi Commixtae Fructus 50% ethanol extract and chlorogenic acid attenuated protein level of MMP3 and mRNA level of MMP13 in TNF𝛼-activated ATDC5 cells. Sorbi commixtae Fructus 50% ethanol extract inhibited the level of PGE2, IL6 and NO in LPS-activated RAW 264.7 cells in dose dependent manner, but chlorogenic acid has no anti-inflammatory effects. Conclusions : These findings demonstrated that Sorbi Commixtae Fructus 50% ethanol extract has chondro-protective and anti-inflammatory effects showing possible therapeutics to ease the symptoms related with osteoarthritis.

• The Korea Journal of Herbology
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• 제37권6호
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• pp.1-8
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• 2022

Objectives : This research aimed to investigate chondro-protective and anti-inflammatory effects of Caraganae Sinicae Flos 50% ethanol extract and its compound, tilianin. Methods : Caraganae Sinicae Flos was extracted with 50% ethanol. Tilianin in Caraganae Sinicae Flos 50% ethanol extract was quantified by HPLC analysis method. To investigate chondro-protective effects of Caraganae Sinicae Flos 50% ethanol extract, ATDC5 chondrogenic cells were co-treated with Caraganae Sinicae Flos 50% ethanol extract (or tilianin) and tumor necrosis factor-𝛼 (TNF𝛼) for 24 hours. After treatement for 24 hours, media supernatant was used for quantifying protein level of matrix metalloproteinase-3 (MMP3) by ELISA and harvested cells were used for analyzing mRNA expression level of matrix metalloproteinase-13 (MMP13) by reverse transcription PCR. To identify anti-inflammatory effects of Caraganae Sinicae Flos 50% ethanol extract, RAW 264.7 macrophage cells were co-treated with Caraganae Sinicae Flos 50% ethanol extract (or tilianin) and lipopolysaccharide (LPS) for 24 hours. media was used for quantifying the level of prostaglandin E2 (PGE2), interleukin-6 (IL6) by ELISA and nitric oxide by Griess reagent asssay. Results : Caraganae Sinicae Flos 50% ethanol extract and tilianin attenuated protein level of MMP3 and mRNA expression level of MMP13 in TNF𝛼-activated ATDC5 cells. Caraganae Sinicae Flos 50% ethanol extract inhibited the level of PGE2, IL6 and NO in LPS-activated RAW 264.7 cells in dose dependent manner, though tilianin inhibited PGE2 only. Conclusions : These results presented that Caraganae Sinicae Flos 50% ethanol extract could be used as natural medicines for osteoarthritis.

• Asian Pacific Journal of Cancer Prevention
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• 제14권11호
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• pp.6869-6874
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• 2013

Background: Vascular endothelial growth factor and matrix metalloproteinases are two important factors for angiogenesis associated with breast cancer growth and progression. The present study was aimed to examine the effects of tamoxifen and tranilast drugs singly or in combination on proliferation of breast cancer cells and also to evaluate VEGF and MMP-9 expression and VEGF secretion levels. Materials and Methods: Human breast cancer cell lines, MCF-7 and MDA-MB-231, were treated with tamoxifen and/or tranilast alone or in combination and percentage cell survival and proliferative activity were evaluated using LDH leakage and MTT assays. mRNA expression and protein levels were examined by real-time RT-PCR and ELISA assay, respectively. Results: LDH and MTT assays showed that the combined treatment of tamoxifen and tranilast resulted in a significant decrease in cell viability and cell proliferation compared with tamoxifen or tranilast treatment alone, with significant decrease in VEGF mRNA and protein levels. We also found that tamoxifen as a single agent rarely increased MMP-9 expression. A decrease in MMP-9 expression was seen after treatment with tranilast alone and in the combined treatment MMP-9 mRNA level was decreased. Conclusions: This combination treatment can able to inhibit growth, proliferation and angiogenesis of breast cancer cells.

• The Korea Journal of Herbology
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• 제31권4호
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• pp.53-60
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• 2016

Objective : The purpose of this study was to investigate anti-aging and antioxidant effects of extracts of Nelumbo nucifera leaves (NN-L) using ethanol on skin .Methods : Each part of leaves(NN-L), flowers(NN-F) and stem(NN-S) was extracted with 70% ethanol. We performed radical scavenging assay(DPPH, ABTS+, Superoxide anion radical), elastase inhibition assay, collagenase inhibition assay. NN-L extracts were tested for cell viability(MTT assay), MMP-1 inhibition and MMP-1 protein expression on CCD-986sk cells (human fibroblast line).Results : Recently, many studies have reported that elastin is also involved in inhibiting or repairing wrinkle formation, although collagen is a major factor in the skin wrinkle formation. We measured its free radical scavenging activity, elastase inhibitory activity and expression of MMP-1 (matrix metalloprotease-1) in human fibroblast cells. Among the parts of Nelumbo nucifera, NN-L showed the highest antioxidant activities and in radical scavenging. DPPH, ABTS+ and Superoxide anion radical scavenging activity of NN-L at concentration of 1,000 μg/mL were 91.43%, 99.31% and 73.7% respectively. In vitro elastase and collagenase inhibition effects of NN-L at concentration of 1,000 μg/mL was 42.8% and 55.3% respectively. The ethanol extract of NN-L showed cell viability of 95.4% in 50 μg/mL concentration. In addition, The results from Western blot assay showed that NN-L decreased the expression of MMP-1 protein in a dose-dependent manner (by up to 35.0% at 50 μM).Conclusion : The findings suggest that the NN-L great potential as a cosmeceutical ingredient with antioxidant and anti-wrinkling effects.