• Journal of Oral Medicine and Pain
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• v.36 no.3
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• pp.147-160
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• 2011

Eugenol (4-allyl-2-methoxyphenol) is a natural phenolic constituent extensively used in dentistry as a component of zinc oxide eugenol cement and is applied to the mouth environment. Chios gum mastic (CGM) is a resinous exudate obtained from the stem and the main leaves of Pistacia lenticulus tree native to Mediterranean areas. This study was undertaken to investigate the synergistic apoptotic effect of co-treatment with a natural product, CGM and natural phenolic compound, eugenol on SCC25 human tongue squamous cell carcinoma cell line. To investigate whether the co-treatment with eugenol and CGM compared to each single treatment efficiently reduces the viability of SCC25 cells, MTT assay was conducted. Induction and augmentation of apoptosis were confirmed by Hoechst staining, TUNEL staining and DNA hypoploidy. Westen blot analysis and immunofluorescent staining were performed to study the alterations of the expression level and the translocation of apoptosis-related proteins in co-treatment. In this study, co-treatment of with eugenol and CGM on SCC25 cells showed several lines of apoptotic manifestation such as nuclear condensations, DNA fragmentation, the increase and decrease of Bax and Bcl-2, decrease of DNA content, the release of cytochrome c into cytosol, translocation of AIF and DFF40 (CAD) onto nuclei, and activation of caspase-3, caspase-6 caspase-7, caspase-9, PARP, Lamin A/C and DFF45 (ICAD) whereas each single treated SCC25 cells did not show or very slightly these patterns. Although the single treatment of 40 ${\mu}g$/ml CGM and 0.5 mM eugenol for 24 h did not induce apoptosis, the co-treatment of these reagents prominently induced apoptosis. Therefore our data provide the possibility that combination therapy with CGM and eugenol could be considered as a novel therapeutic strategy for human oral squamous cell carcinoma.

• Tuberculosis and Respiratory Diseases
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• v.44 no.6
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• pp.1353-1365
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• 1997

Background : Tumor necrosis factor(TNF) has been considered as an important candidate for cancer gene therapy based on its potent anti-tumor activity. However, since the efficiency of current techniques of gene transfer is not satisfactory, the majority of current protocols is aiming the in vitro gene transfer to cancer cells and re-introducing genetically modified cancer cells to host. In the previous study, it was shown that TNF-sensitive cancer cells transfected with TNF-$\alpha$ cDNA would become highly resistant to TNF, and the probability was shown that the acquired resistance to TNF might be associated with synthesis of some protective protein. Understanding the mechanisms of TNF -resistance in TNF-$\alpha$ cDNA transfected cancer cells would be. an important step for improving the efficacy of cancer gene therapy as well as for better understandings of tumor biology. This study was designed to evaluate the role of MnSOD, an antioxidant enzyme, in the acquired resistance to TNF of TNF-$\alpha$ cDN A transfected cancer cells. Method : We transfected TNF-$\alpha$ c-DNA to WEHI164(murine fibrosarcoma cell line), NCI-H2058(human mesothelioma cell line), A549(human non-small cell lung cancer cell line), ME180(human cervix cancer cell line) cells using retroviral vector(pLT12SN(TNF)) and confirm the expression of TNF with PCR, ELISA, MIT assay. Then we determined the TNF resistance of TNF-$\alpha$ cDNA transfected cells(WEHI164-TNF, NCIH2058-TNF, A549-TNF, ME180-TNF) and the changes of MnSOD mRNA expressions with Northern blot analysis. Results : The MnSOD mRNA expressions of parental cells and genetically modified cells of WEHI164 and ME180 cells(both are naturally TNF sensitive) were not significantly different The MnSOD mRNA expressions of genetically modified cells of NCI-H2058 and A549(both are naturally TNF resistant) were higher than those of the parental cells, while those of parental cells with exogenous TNF were also elevated. Conclusion : The acquired resistance to TNF after TNF-$\alpha$ cDNA transfection may not be associated with the change in the MnSOD expression, but the difference in natural TNF sensitivity of each cell may be associated with the level of the MnSOD expression.

• Tuberculosis and Respiratory Diseases
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• v.58 no.4
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• pp.375-384
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• 2005

Background and Aims : Pre-induction of heat shock protein 70 (HSP70) is known to effectively attenuate the lipopolysaccharide (LPS)-induced inflammatory response in lung tissue. However, it is unclear if HSP70 induction after LPS exposure attenuates the subsequent LPS-induced inflammatory response in alveolar epithelial cells. This study examined the effects of HSP70 induction after LPS exposure on the IL-6 production and the cell viability after a subsequent LPS challenge in murine alveolar epithelial cells, and investigated whether or not HSP70 itself may be involved in those effects. Methods : Murine alveolar epithelial cells were cultured and divided into two groups; the Non-Pre-LPS group without a LPS pre-treatment and the Pre-LPS group with a LPS pre-treatment. Each group was subdivided into the following four subgroups: subgroup C (control), subgroup Q (quercetin), subgroup HSP70 (HSP70 induction), and subgroup HSP70-Inh (HSP70 inhibition). HSP70 expression, which was induced by sodium arsenite and inhibited by quercetin, was analyzed by western blot analysis. The IL-6 levels in the culture supernatant were measured by ELISA, and the cell viability was measured using a simplified MTT assay. Results : The IL-6 levels were lower in subgroup HSP70 than in subgroup C (P<0.01), and were higher in subgroup HSP70-Inh than in subgroup HSP70 in both the Non-Pre-LPS and Pre-LPS groups (P<0.05, P<0.01). The cell viability tended to decrease in the Pre-LPS group compared with the Non-Pre-LPS group. While the cell viability was higher in subgroups Q, HSP70, and HSP70-Inh than in subgroup C in the Non-Pre-LPS group (P<0.05, P<0.05, P<0.01), there was no difference in cell viability among the subgroups in the Pre-LPS group. Conclusion : HSP70 induction after a LPS pre-treatment in murine alveolar epithelial cells inhibits the subsequent LPS-induced IL-6 production without affecting the cell viability, and HSP70 by itself may play an important role in this proccess.

• Korean Journal of Medicinal Crop Science
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• v.15 no.1
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• pp.30-37
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• 2007

It is well-known that Korean mistletoe (Viscum album) extract has an immune activity and anticancer effect. In this study, Korean mistletoe extract, M11C (non-lectin components), was used to examine whether this extract might activate human peripheral monocyte to produce tumor necrosis $factor-\alpha$ $(TNF-\alpha)$. To examine the effect of M11C on the production of $TNF-\alpha$ from monocyte, the monocyte were stimulated by the M11C, and then collected the supernatant (M11C stimulated monocyte-conditioned media; MCM). MCM was treated to the $TNF-\alpha$ sensitive L929 cells, and then L929 cytotoxicity was measured by means of MTT. MCM had cytotoxic effect on L929. And the cytotoxic effect of MCM on L929 was almost abolished by $anti-TNF-\alpha$ antibody. These data indicated that MCM contained $TNF-\alpha$, suggesting the $TNF-\alpha$ generation from M11C-stimulated monocyte. This suggestion was confirmed from the data that $TNF-\alpha$ was highly detected in MCM by immunoblotting technique. M11C effect on $TNF-\alpha$ production from monocyte was in the dose and stimulating time dependent manners. Also the effect of M11C on the expression of $TNF-\alpha$ mRNA from monocyte was shown in the dose and stimulating time dependent manners. As a result, Korean mistletoe extract, M11C, could be used for an immunostimulator.

• Korean Journal of Food Science and Technology
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• v.46 no.5
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• pp.609-615
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• 2014

The cytoprotective effect of Moringa oleifera Lam. (drumstick tree) on neuronal cells was investigated to confirm the physiological benefits associated with this natural food resource. First, the drumstick tree extract was chemically analyzed to determine inherent nutritional constituents. Calcium and potassium were identified as the major mineral constituents, and palmitic acid (C16:0, 16.33%) and gadoleic acid (C20:01, 66.34%) were detected as the major fatty acids. Moreover, drumstick tree extract contained 94.78 mg/100 g vitamin E and 112.61 mg/100 g niacin. PC12 cells were used to study the cytoprotective effects of drumstick tree extract. Intracellular accumulation of reactive oxygen species was significantly reduced when $H_2O_2$ treated-neuronal cells were cultured in a medium containing the methanolic extract of drumstick tree, compared to cells treated with only $H_2O_2$. Cell viability assay using MTT showed that the extract protected cells against $H_2O_2$-induced neurotoxicity and inhibited LDH leakage from the cell membrane. Caspase assay showed that the extract exerted cytoprotective effect against apoptosis. Consequently, these data suggest that drumstick tree is a useful natural resource with positive effects on human health.

• Food Science and Preservation
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• v.15 no.5
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• pp.743-748
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• 2008

Amyloid $\beta$ peptide ($A{\beta}$) is known to increase oxidative stress in nerve cells, leading to apoptosis that is characterized by free radical formation and lipid peroxidation. Neurodegenerative diseases such as Alzheimer's disease (AD) are characterized by large deposits of $A{\beta}$ in the brain. In our study, neuronal protective effects of green tea, along with water activity (0.813), and leaf storage periods (fresh leaf, or leaf stored for up to 4 weeks) were investigated. We measured protective effects against $A{\beta}$-induced cytotoxicity in neuron-like PC12 cells. Powdered green tea was extracted with distilled water at $70^{\circ}C$ for 5 min, and this extract was freeze-dried and stored at $-20^{\circ}C$ until use. In cell viability assays using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT), the fresh extract, and that obtained after 1 week of leaf storage, showed the best protective effects against $A{\beta}$-induced neurotoxicity. As oxidative stress causes membrane breakdown, the protective effect of green tea extracts was investigated using lactate dehydrogenase (LDH) and trypan blue exclusion assays. LDH release into the medium was inhibited (by 20-25%) in all tests. In addition, all green tea extracts (fresh, or stored before extraction for up to 4 weeks) showed better cell protective effects ($93.3{\pm}1.8-96.2{\pm}2.4$) than did vitamin C ($91.0{\pm}1.6$), used as a positive control. The results suggest that effectiveness of green tea extracts falls with prolonged leaf storage.

• Journal of Life Science
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• v.31 no.9
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• pp.827-833
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• 2021

Dendropanax morbiferus leaves (DPL) has been used as a medicine since ancient times in various diseases such as inflammation, diabetes, and cancer. In particular, it has been found to have anticancer effects on several types of cancer cells, but the anticancer effect on breast cancer cells SK-BR-3 has not yet been revealed. Therefore, in this study, DPL caused proliferation inhibition in breast cancer cells SK-BR-3 and the anticancer effect by inducing apoptosis was confirmed, through an in vitro experiment. In order to examine the effect of DPL on cell viability, MTT assay was performed to confirm a significant decrease in the concentration of cell viability. DAPI staining was performed to examine the effect of DPL on cellular morphological changes and increase of apoptotic bodies. To supplement this, an increase in the apoptosis rate was also confirmed through flow cytometry after staining with annexin V/PI. Western blot was performed to confirm apoptosis-related proteins. DPL increased the expression of Cleaved-PARP, Bax whereas decreased the expression of Bcl-2. Changes in the expression levels of MAPK pathway proteins p-ERK1/2, p-JNK, and p-p38 were also confirmed, and a significant increase in p-p38 was observed. These results indicated that DPL induced apoptosis, through p-p38 MAPK signal pathway in SK-BR-3 breast cancer cells.

• Journal of Life Science
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• v.29 no.2
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• pp.173-180
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• 2019

Amyloid ${\beta}$-protein ($A{\beta}$) is the principal component of senile plaques characteristic of Alzheimer's disease (AD) and elicits a toxic effect on neurons in vitro and in vivo. Many environmental factors, including antioxidants and proteoglycans, modify $A{\beta}$ toxicity. It is worthwhile to isolate novel natural compounds that could prove therapeutic for patients with AD without causing detrimental side effects. In this study, we investigated the in vitro neuroprotective effects of the ethyl acetate fraction of methanol extract of Ophiophogon japonicas (OJEA fraction). We used an MTT reduction assay to detect protective effects of the OJEA fraction on $A{\beta}_{25-35}$-induced cytotoxicity to PC12 cells. We also used a cell-based ${\beta}$-secretase assay system to investigate the inhibitory effect of the OJEA fraction on ${\beta}$-secretase activity. In addition, we performed an in vitro lipid peroxidation assay to evaluate the protective effect of the OJEA fraction against oxidative stress induced by $A{\beta}_{25-35}$ in PC12 cells. The OJEA fraction had strong protective effects against $A{\beta}_{25-35}$-induced cytotoxicity to PC12 cells and was strongly inhibitory to ${\beta}$-secretase activity, which resulted in the attenuation of $A{\beta}$ generation. In addition, the OJEA fraction significantly decreased malondialdehyde (MDA) content, which is induced by the exposure of PC12 cells to $A{\beta}_{25-35}$. Our results suggested that the OJEA fraction contained active compounds exhibiting a neuroprotective effect on $A{\beta}$ toxicity.

• Journal of the Korean Society of Food Science and Nutrition
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• v.41 no.12
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• pp.1708-1715
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• 2012

The purpose of this study was to investigate whether water extracts of Sparassis crispa (SC) have anti-obesity effects. Treatment of mature adipocytes with SC caused a decrease in lipid accumulation (assessed by Oil Red O staining) and an increase in glycerol release. Mice were induced to obesity by a high fat diet (45% fat in total kcal) and experimental groups were treated with two different dosages of SC extracts, a low SC (LSC, 100 mg/kg/day) or high SC (HSC, 300 mg/kg/day). SC extracts were administered by gavages for 10 weeks in the experimental groups, while the control group was fed with distilled water. The body weight gain of mice fed SC was significantly reduced (11.88% lower in LSC, 14.54% lower in HSC) compared to the control group. Additionally, there were significantly reduced serum levels of triglycerides (13.57% lower in LSC, 19.46% lower in HSC), total cholesterol (32.22% lower in LSC, 24.67% lower in HSC) and glucose (28.85% lower in LSC, 25.82% lower in HSC) in mice fed SC compared to the control group. Hepatic triglycerides in mice fed SC were lower (9.68% lower in LSC, 14.24% lower in HSC) than the control group and total cholesterol levels were also lower in mice fed SC (38.72% lower in LSC, 35.20% in HSC). These results demonstrate that the water extract of SC may enhance lipolysis and up-regulate the expression of lipolytic enzymes in vitro and reduce body weight in vivo. These significant effects were found for both low and high doses of SC treatment, and suggest SC can be used as potential therapeutic substances for the prevention and treatment of obesity.

• Journal of the Korean Society of Food Science and Nutrition
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• v.43 no.1
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• pp.47-54
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• 2014

The aim of this study is to evaluate the effects of green tea polyphenol (GTP) on anticancer treatment with cisplatin (CP), using both an in vitro cell culture model and an in vivo mouse model of established breast tumor. Mouse breast cancer cells (EMT6) were treated with or without GTP and CP followed by determination of the cell viability using an MTT assay. The relative cell viability of CP treated EMT6 cells was 96% at a 20 ${\mu}g/mL$ concentration of cisplatin; however, in combination with GTP (50 ${\mu}g/mL$), the cell viability decreased to 20% at the same concentration of CP (20 ${\mu}g/mL$). For the in vivo study, EMT6 cells were inoculated into Balb/c mice for the establishment of a tumor-bearing mice model. The tumor-bearing mice were treated with CP (5 mg/kg. i.p.) with or without dietary GTP (0.2% drinking water). Tumor growth was monitored by a measurement of tumor size using a digital caliper, and nephrotoxicity was determined by enzymatic and histological examinations. The levels of p53 and caspase-3 in tumor tissues were examined by a Western blot. In tumor-bearing mice treated with GTP plus CP, the increment of tumor volume showed a significant reduction, compared with CP or GTP alone. The levels of p53 and cleaved caspase-3 (caspase-3/p17) in tumor tissues of tumor-bearing mice were increased by CP and GTP compared to CP alone. In CP treated tumor-bearing mice, ${\gamma}$-glutamyltranspeptidase (GGT) and alkaline phosphatase (AP) activities were decreased, and marked tubular necrosis and dilatation were observed in the kidney. CP-induced enzymatic and histopathological changes in the kidney of tumor-bearing mice were reduced by combinations of GTP with CP. The results of these experiments demonstrated that dietary GTP has a potentiating effect on CP anti-tumor activity and a protective effect against CP-induced renal dysfunction. Therefore, GTP may be used as a modulator in anticancer treatment with CP.