• Korean Journal of Food Science and Technology
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• v.45 no.2
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• pp.257-261
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• 2013

Rat pheochromocytoma cells (PC12) and mice were utilized as in vitro and in vivo models to determine the neuroprotective effects of a 70% acetone extract of black soybean seed coat (BSSCE). BSSCE showed higher total phenolic contents than other extracts. Intracellular reactive oxygen species accumulation from $H_2O_2$ treatment of PC12 cells was significantly reduced when BSSCE was present in the media compared to PC12 cells treated with $H_2O_2$ only. The 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-tetrazolium-bromide (MTT) reduction assay and lactate dehydrogenase assay also showed significantly increased protective effects in PC12 cells. In addition, BSSCE improved the in vivo cognitive ability against amyloid beta peptide-induced neuronal deficits.

• Asian Pacific Journal of Cancer Prevention
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• v.16 no.8
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• pp.3383-3387
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• 2015

Portulaca oleracea (Family: Portulacaceae), is well known for its anti-inflammatory, antioxidative, anti-bacterial, and anti-tumor activities. However, cytotoxic effects of seed oil of Portulaca oleracea against human liver cancer (HepG2) and human lung cancer (A-549) cell lines have not been studied previously. Therefore, the present study was designed to investigate the cytotoxic effects of Portulaca oleracea seed oil on HepG2 and A-549 cell lines. Both cell lines were exposed to various concentrations of Portulaca oleracea seed oil for 24h. After the exposure, percentage cell viability was studied by (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) (MTT), neutral red uptake (NRU) assays, and cellular morphology by phase contrast inverted microscopy. The results showed a concentration-dependent significant reduction in the percentage cell viability and an alteration in the cellular morphology of HepG2 and A-549 cells. The percentage cell viability was recorded as 73%, 63%, and 54% by MTT assay and 76%, 61%, and 50% by NRU assay at 250, 500, and $1000{\mu}g/ml$, respectively in HepG2 cells. Percentage cell viability was recorded as 82%, 72%, and 64% by MTT assay and 83%, 68%, and 56% by NRU assay at 250, 500, and $1000{\mu}g/ml$, respectively in A-549 cells. The 100 $100{\mu}g/ml$ and lower concentrations were found to be non cytotoxic to A-549 cells, whereas decrease of 14% and 12% were recorded by MTT and NRU assay, respectively in HepG2 cells. Both HepG2 and A-549 cell lines exposed to 250, 500, and $1000{\mu}g/ml$ of Portulaca oleracea seed oil lost their normal morphology, cell adhesion capacity, become rounded, and appeared smaller in size. The data from this study showed that exposure to seed oil of Portulaca oleracea resulted in significant cytotoxicity and inhibition of growth of the human liver cancer (HepG2) and human lung cancer (A-549) cell lines.

• Journal of Life Science
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• v.28 no.11
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• pp.1316-1320
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• 2018

The effects of a green tea extract (GTE) were examined using the MCF-7 human breast cancer cell line. Cell viability assays using 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assays revealed that GTE had a significant cytotoxic effect on MCF-7 cells, depending on the concentration of GTE. Western blotting of p53 and its related proteins, p21/cip1 and CDK2, after GTE treatment revealed that a significant and concentration dependent increase in p53 protein in response to GTE. The levels of p21/cip1 proteins were also increased at low GTE concentrations were significantly increased even at the highest GTE concentrations. However, the level of CDK2 was significantly decreased by treatment with high concentrations of GTE. These results indicate that treatment with GTE increased the p53 level in MCF-7 cells, and this activation of p53 markedly elevated the levels of p21/cip1proteins, which, in turn, inhibited CDK2 expression in the MCF-7 cells. The inhibition of CDK2 expression might then affect cell cycle progression. Subsequent FACS analysis indicated that GTE treatment the gradually increased progression of the MCF-7 to the G1 phase. These results clearly demonstrate that the anti-tumor effect of GTE in MCF-7 cells is regulated by p53 arrest of the MCF-7 cells at the G1 stage of cell cycle.

• KOREAN JOURNAL OF CROP SCIENCE
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• v.56 no.1
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• pp.80-87
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• 2011

Panax ginseng has been used as a traditional medicine for several centuries in Korea. A laboratory experiment using methanol extracts of freeze-dried leaves and roots in the different ages of P. ginseng was conducted to determine the content of phenolics and flavonoids, antioxidant activity and cytotoxicity. The results indicate that the total phenolics level [mg ferulic acid equivalents (FAE) $kg^{-1}$ DW] was higher in leaves (22.0 to 76.3 mg $kg^{-1}$) than roots (19.0 to 28.3 mg $kg^{-1}$) of P. ginseng. The total content of phenolics in roots increased with increase in age of P. ginseng from one to six years. However, the content of phenolics in P. ginseng leaf decreased with the increase in age. Total flavonoid [mg naringin equivalents $kg^{-1}$ DW] was more detected in the leaves (30.3 to 138.6 mg $kg^{-1}$) than in the roots (0.0 to 10.6 mg $kg^{-1}$) of P. ginseng. The total flavonoid level in leaves decreased with increase in age of P. ginseng. The antioxidant potential of the methanol extracts from the plants dose-dependently increased. DPPH free radical scavenging activity was higher in leaves (36.9 to 82.8%) than in roots (14.8 to 39.4%), and in young plants than in old ones. According to 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay, the methanol extracts from 5 year-root part showed the highest cytotoxicity against Calu-6, followed by 2 year- and 3 year-roots. However, the methanol extracts from 6 year- and 4 year-roots had lower cytotoxicity. Total phenolics content in both leaves and roots was highly correlated with the DPPH radical scavenging ($r^2=0.7366$ to 0.7870) and nitrite scavenging ($r^2=0.5604$ to 0.8794) activities, suggesting that they contribute to the antioxidant properties of the P. ginseng plants.

• Journal of the Korean Society of Food Science and Nutrition
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• v.45 no.8
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• pp.1107-1113
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• 2016

The purpose of this study was to investigate the effects of catechu water extract on adipogenesis in 3T3-L1 adipocytes. 3T3-L1 preadipocytes were differentiated with adipogenic regents by incubation for 9 days in the absence or presence of catechu extract ranging from $1{\sim}200{\mu}g/mL$. The effect of catechu extracts on cell proliferation of 3T3-L1 preadipocytes was investigated using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay. The effect of catechu extracts on 3T3-L1 differentiation was examined by measuring intracellular lipid droplet and triglyceride contents. These results were obtained from preadipocyte proliferation and adipocyte differentiation of 3T3-L1. Catechu extracts inhibited lipid accumulation and remarkably decreased triglyceride contents in 3T3-L1 preadipocytes at a concentration showing no cytotoxicity. The anti-adipogenic effects of catechu appeared to be mediated by significant down-regulation of expression of peroxisome proliferator-activated receptor ${\gamma}$, CCAAT/enhancer-binding protein ${\alpha}$, and sterol regulatory element-binding protein 1c proteins apart from expression of hormone-sensitive lipase. We suggest that catechu extracts significantly inhibit adipogenesis and can be used for regulation of obesity.

• Asian-Australasian Journal of Animal Sciences
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• v.32 no.2
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• pp.257-264
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• 2019

Objective: Dairy cattle nutrient requirement systems acknowledge amino acid (AAs) requirements in aggregate as metabolizable protein (MP) and assume fixed efficiencies of MP used for milk protein. Regulation of mammary protein synthesis may be associated with AA input and milk protein output. The aim of this study was to evaluate the effect of nanoemulsified methionine and cysteine on the in-vitro expression of milk protein (casein) in bovine mammary epithelial cells (MAC-T cells). Methods: Methionine and cysteine were nonionized using Lipoid S 75 by high-speed homogenizer. The nanoemulsified AA particle size and polydispersity index were determined by dynamic light scattering correlation spectroscopy using a high-performance particle sizer instrument. 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay was performed to determine the cytotoxicity effect of AAs with and without nanoionization at various concentrations (100 to $500{\mu}g/mL$) in mammary epithelial cells. MAC-T cells were subjected to 100% of free AA and nanoemulsified AA concentration in Dulbecco's modified Eagle medium/nutrient mixture F-12 (DMEM/F12) for the analysis of milk protein (casein) expression by the quantitative reverse transcription polymerase chain reaction method. Results: The AA-treated cells showed that cell viability tended to decrease (80%) in proportion to the concentration before nanogenesis, but cell viability increased as much as 90% after nanogenesis. The analysis of the expression of genetic markers related to milk protein indicated that; ${\alpha}_{s2}$-casein increased 2-fold, ${\kappa}$-casein increased 5-fold, and the amount of unchanged ${\beta}$-casein expression was nearly doubled in the nanoemulsified methionine-treated group when compared with the free-nanoemulsified methionine-supplemented group. On the contrary, the non-emulsified cysteine-administered group showed higher expression of genetic markers related to milk protein ${\alpha}_{s2}$-casein, ${\kappa}$-casein, and ${\beta}$-casein, but all the genetic markers related to milk protein decreased significantly after nanoemulsification. Conclusion: Detailed knowledge of factors, such nanogenesis of methionine, associated with increasing cysteine and decreasing production of genetic markers related to milk protein (casein) will help guide future recommendations to producers for maximizing milk yield with a high level of milk protein casein.

• Journal of Periodontal and Implant Science
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• v.48 no.1
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• pp.34-46
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• 2018

Purpose: The purpose of this study was to evaluate the effects of 1,25-dihydroxyvitamin $D_3$ on the proliferation, differentiation, and matrix mineralization of MC3T3-E1 osteoblast-like cells in vitro. Methods: MC3T3-E1 osteoblastic cells and 1,25-dihydroxyvitamin $D_3$ were prepared. Cytotoxic effects and osteogenic differentiation were evaluated using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay, alkaline phosphatase (ALP) activity assay, ALP staining, alizarin red S staining, and reverse transcription-polymerase chain reaction (RT-PCR) for osteogenic differentiation markers such as ALP, collagen type I (Col-I), osteocalcin (OCN), vitamin D receptor (VDR), and glyceraldehyde 3-phosphate dehydrogenase. Results: The MTT assay showed that 1,25-dihydroxyvitamin $D_3$ did not inhibit cell growth and that the rate of cell proliferation was higher than in the positive control group at all concentrations. ALP activity was also higher than in the positive control group at low concentrations of 1,25-dihydroxyvitamin $D_3$ ($10^{-10}$, $10^{-12}$, and $10^{-14}M$). RT-PCR showed that the gene expression levels of ALP, Col-I, OCN, and vitamin D receptor (VDR) were higher at a low concentration of 1,25-dihydroxyvitamin $D_3$ ($10^{-12}M$). Alizarin red S staining after treatment with 1,25-dihydroxyvitamin $D_3$ ($10^{-12}M$) showed no significant differences in the overall degree of calcification. In contrast to the positive control group, formation of bone nodules was induced in the early stages of cell differentiation. Conclusions: We suggest that 1,25-dihydroxyvitamin $D_3$ positively affects cell differentiation and matrix mineralization. Therefore, it may function as a stimulating factor in osteoblastic bone formation and can be used as an additive in bone regeneration treatment.

• The Korea Journal of Herbology
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• v.29 no.2
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• pp.39-45
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• 2014

Objectives : The purpose of this study was to investigate antiaging and antioxidant effects on cultured human skin fibroblast with 80% ethanol extracts of plants including of stem of Dendropanax morbifera, Corni fructus and Lycii Fructus. Methods : An ethanol extract of three medicinal plants including stem of Dendropanax morbifera, Corni fructus and Lycii Fructus. Extracts were assessed to determine the mechanism of antioxidant and antiaging activities. Antioxidant activity of extract was evaluated by two different assays as 2,2-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging and super oxide dismutase (SOD) like activities. These extracts were tested for cell viability on HS68 skin fibroblast by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-tetrazolium bromide (MTT) assay. We investigated the effects of Ultraviolet-B irradiation on cytotoxicity, type 1 collagen, elastin level and oxidative damage in cultured human skin fibroblast (HS68). 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. Results : The extracts obtained dose-dependently increased the scavenging activity on DPPH radical scavenging activity and SOD like activity. The extracts of complex herbal medicine showed low cytotoxicity as more than 100% cell viability in 100ppm/ml concentration. HS68 fibroblasts were survived 70% at 120 $mJ/cm^2$ UVB irradiation and treated tumor necrosis factor (TNF)-alpha. The levels of aging factors and cytotoxicity were decreased by ethanol extract of complex herbal medicine. Conclusions : These results suggest that ethanol extracts of complex medicinal plants of including of stem of Dendropanax morbifera, Corni fructus and Lycii Fructus may have value as the potential antioxidant and antiaging medicinal plant.

• KOREAN JOURNAL OF CROP SCIENCE
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• v.58 no.1
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• pp.20-27
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• 2013

Proximate composition, total phenolics and total flavonoids level, DPPH radical scavenging activity, and cytotoxicity were determined in the methanol extracts of different plant parts of Youngia sonchifolia at reproductive growth stage. Crude protein and crude fat were present as the highest amount in flowers, and crude fiber in the stems and roots. The highest content of phenolics [mg ferulic acid equivalents (FAE) $kg^{-1}$ dry weight (DW)] was found in flowers (highest) and followed by leaves, stems and roots (lowest). Flavonoids [mg rutin equivalents $kg^{-1}$ DW] level, however, showed the highest in leaf extracts and lowest in root extracts. The antioxidant potential of the methanol extracts from the plants dose-dependently increased DPPH (1,1-diphenyl-2-picrylhydrazyl) free radical scavenging activity (%). DPPH radical scavenging activity were highest in root extracts ($IC_{50}=1,135.6\;mg\;kg^{-1}$) and followed by leaf, stem and flower extracts. By way of MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide) assay, methanol extracts of roots showed the highest anticancer activity on human cancer cell line Calu-6 for human pulmonary carcinoma ($IC_{50}=196.3\;mg\;kg^{-1}$) and HCT-116 for human colon carcinoma ($IC_{50}=623.6\;mg\;kg^{-1}$).

• Journal of Veterinary Science
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• v.21 no.5
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• pp.80.1-80.13
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• 2020

Background: In suckling piglets, transmissible gastroenteritis virus (TGEV) causes lethal diarrhea accompanied by high infection and mortality rates, leading to considerable economic losses. This study explored methods of preventing or inhibiting their production. Bovine antimicrobial peptide-13 (APB-13) has antibacterial, antiviral, and immune functions. Objectives: This study analyzed the efficacy of APB-13 against TGEV through in vivo and in vitro experiments. Methods: The effects of APB-13 toxicity and virus inhibition rate on swine testicular (ST) cells were detected using 3-(4,5-dimethylthiazol-2-yl)-2, 5-diphenyl tetrazolium bromide (MTT). The impact of APB-13 on virus replication was examined through the 50% tissue culture infective dose (TCID₅₀). The mRNA and protein levels were investigated by real-time quantitative polymerase chain reaction and western blot (WB). Tissue sections were used to detect intestinal morphological development. Results: The safe and effective concentration range of APB-13 on ST cells ranged from 0 to 62.5 ㎍/mL, and the highest viral inhibitory rate of APB-13 was 74.1%. The log₁₀TCID₅₀ of 62.5 ㎍/mL APB-13 was 3.63 lower than that of the virus control. The mRNA and protein expression at 62.5 ㎍/mL APB-13 was significantly lower than that of the virus control at 24 hpi. Piglets in the APB-13 group showed significantly lower viral shedding than that in the virus control group, and the pathological tissue sections of the jejunum morphology revealed significant differences between the groups. Conclusions: APB-13 exhibited good antiviral effects on TGEV in vivo and in vitro.