• Journal of Animal Science and Technology
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• v.55 no.4
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• pp.237-247
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• 2013

This study was to examine single or combined in vitro effects of environmental endocrine disruptors on boar sperm characteristics, oxidative stress damage in sperm and development of porcine IVF embryos. Addition of various concentration of NP (10, 20, $30{\mu}M$), DBP (10, 50, $100{\mu}M$) and BPA (1, 5 or $10{\mu}g/ml$) on boar sperm characteristics such as percentages of sperm motility, viability, membrane integrity and mitochondrial activity were dose-dependently decreased within 3, 6 or 9 hr incubation period (p<0.05). The overall detrimental effects increased with incubation time increasement. NP, DBP and BPA showed the detrimental effects on sperm membrane and mitochondria of energy production organelles affecting cell viability with the dependancy of dose and incubation time. In combination effects, NP ($10{\mu}M$) + DBP ($10{\mu}M$) significantly decreased boar general sperm characteristics for 3 or 6 hr incubation period compared with control (p<0.05). When both of NP and DBP concentrations (NP; $30{\mu}M$, DBP; $100{\mu}M$) increase, the detrimental effects on sperm characteristics were larger than those of low concentration combination (p<0.05). The inhibitory effects of NP ($30{\mu}M$) + BPA ($10{\mu}g/ml$) on sperm characteristics were larger than those of NP ($10{\mu}M$) + BPA ($1{\mu}g/ml$) (p<0.05). DBP ($100{\mu}M$) + BPA ($10{\mu}g/ml$) decreased sperm characteristics compared with the low concentration combination (DBP $10{\mu}M$ + BPA $1{\mu}g/ml$, p<0.05). This result indicates the detrimental effects of both chemicals on sperm characteristics were dose dependent. Addition of NP ($30{\mu}M$) + DBP ($100{\mu}M$), NP ($30{\mu}M$) + BPA ($10{\mu}g/ml$), DBP ($10{\mu}M$) + BPA ($1{\mu}g/ml$) or DBP ($100{\mu}M$) + BPA ($10{\mu}g/ml$) significantly increased lipid peroxidation for 3 or 6 hr incubation period (p<0.05) compared with no addition control. NP (${\geq}20{\mu}M$) decreased the percentages of IVF embryo development from morulae and blastocyst stages (p<0.05) and its detrimental effects were dose-dependant. BPA 0, 1, 5 or $10{\mu}g/ml$ decreased significantly and dose-dependently the percentage of morulae plus and blastocysts (p<0.05). Combinations of DBP ($100{\mu}M$) plus NP ($30{\mu}M$) and DBP ($100{\mu}M$) plus BPA ($10{\mu}g/ml$) did not affect on morulae and blastocyst development, but NP ($30{\mu}M$) plus BPA ($10{\mu}g/ml$) has significant detrimental effect on embryo development at these stages (p<0.05). These overall results indicate that the partial detrimental effects on boar sperm characteristics and embryo development by NP, DBP, BPA or the combination of these chemicals might be due to the increasement of lipid peroxidation and free radical formation in the cell and there were no specific interaction effects on boar sperm and embryo degeneration among the combined treatments.

• Journal of Embryo Transfer
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• v.19 no.2
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• pp.89-99
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• 2004

Stretch-activated channels (SACs) responds to membrane stress with changes in open probability (Po). They play essential roles in regulation of cell volume and differentiation, vascular tone, and in hormonal secretion. SACs highly present in Xenopus oocytes and Ascidian oocytes are suggested to be involved in the regulation of pH and fluid transport to balance the osmotic pressure, but remain unclear in mammanlian oocytes. This study was investigated to find the presence of SACs in hamster oocytes and to examine their electrophysiological properties. To infer a role of SAC in relation to the development of early stage, we followed up to the stage of two-cell zygote with patch clamp techniques. Single channels were elicited by negative pressure (lower than ­15 cm$H_2O$). Interestingly, SACs were dependent on permeable cations such as $Na^+$ or $K^+$. As permeable cation removed from both sides across the membrane, SAC activity completely disappeared. When permeable cations present only in intracellular compartment, outward currents appeared at positive potentials. In contrast to this, inward currents occurred only at the negative voltage when permeable cation absent in cell interior. These result suggests that SAC carry cations through the nonselective cation channel (NSC channel). Taken together, we found that stretch activated channels present in hamster oocyte and the channel may carry cations through NSC channels. This stretch activated-NSC channels may play physiological role(s) in oocyte growth, maturation, fertilization and embryogenesis in fertilized oocytes to two-cell zygotes of hamster.

• Journal of the Korean Society of Food Science and Nutrition
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• v.45 no.5
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• pp.651-663
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• 2016

Corn silk, Job's tears, Lentinus edodes, and apple peel 70% ethanol extracts (CS, JT, LE, and AP) were studied for their antioxidant activities. CS among all extracts showed the highest antioxidant activities based on total polyphenol and flavonoid contents, 2,2-diphenyl-${\beta}$-picrylhydrazyl and 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonate) radical scavenging activity, and reducing power. Adipocyte differentiation was investigated by Oil Red O staining assay using CS, JT, LE, AP, and extract of developed bread containing corn silk, Job's tears, Lentinus edodes, and apple peel (DB) treated to 3T3-L1 adipocytes. DB1 and DB2 showed anti-adipogenic and antioxidant effects. Triglyceride (TG) accumulation in 3T3-L1 cells was measured, and among the samples tested (CS, JT, LE, and AP), CS was found to have the highest inhibitory activity against TG accumulation of differentiated 3T3-L1 adipocytes and regulated factors associated with adipogenesis. CS suppressed lipid droplet formation and adipocyte differentiation in 3T3-L1 cells in a dose-dependent manner. We examined the effects of CS on the levels of CCAAT-enhancer-binding protein ${\beta}(C/EBP{\beta})$, peroxisome proliferator activated receptor ${\gamma}(PPAR{\gamma})$, and adipocyte-specific lipid binding protein (aP2) mRNA as well as protein levels in 3T3-L1 cells treated with CS at various concentrations (0, 10, 50, and $100{\mu}g/mL$) during adipocyte differentiation and treatment with CS in 3T3-L1 adipocytes down-regulated expression of $PPAR{\gamma}$ and aP2 mRNA. CS also significantly inhibited up-regulation of $C/EBP{\beta}$, $PPAR{\gamma}$, and aP2 proteins during adipocyte differentiation. These data indicate that DBs have anti-adipogenic activity induced by CS in 3T3-L1 preadipocytes, and CS exerts anti-adipogenic activity by inhibiting expression of $C/EBP{\beta}$, $PPAR{\gamma}$, and aP2 signaling pathway in 3T3-L1 adipocytes. JT, LE, and AP had no inhibitory effects on differentiation of 3T3-L1 preadipocytes but displayed strong antioxidant effects. These results suggest that the developed bread may be a health beneficial food that can prevent or treat obesity and diseases induced by oxidative stress.

• Journal of the Korean Society of Food Science and Nutrition
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• v.42 no.3
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• pp.325-334
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• 2013

The objective of this study was to compare the content and metabolic activities between fresh pine needle juice (PNJ) and fermented pine needle juice (FPNJ). A variety of factors were measured, including total phenolic content (TPC), antioxidant activity [DPPH radical scavenging activity (RSA), total radical-trapping antioxidant potential (TRAP), oxygen radical absorbance capacity (ORAC), cellular antioxidant capacity (CAC)], anti-genotoxic activity, ${\alpha}$-glucosidase inhibitory activity, and angiotensin converting enzyme (ACE) inhibitory activity. The TPC was $17.3{\pm}0.2$ and $4.6{\pm}0.0$ mg GAE/g in PNJ and FPNJ, respectively. The DPPH RSA, TRAP, and ORAC values increased in a dose-dependent manner for both PNJ and FPNJ, with significantly higher activities in PNJ than FPNJ. The CAC against AAPH-induced oxidative stress in HepG2 cells was protected by both PNJ and FPNJ. Pretreatment with PNJ and FPNJ in human leukocytes produced significant reductions in $H_2O_2$-induced DNA damage at a concentration of $50{\mu}g/mL$. ${\alpha}$-Glucosidase inhibitory activity was significantly higher in FPNJ than PNJ. The ACE inhibitory activity was about 87.1% and 60.0% in 1:1 diluted PNJ and FPNJ, respectively. This study suggests that the fermentation of PNJ could enhance the regulation of blood glucose metabolism and both PNJ and FPNJ might be a new potential source of natural antioxidant, anti-diabetic, and anti-hypertensive agents applicable to food.

• Korean Journal of Environmental Biology
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• v.24 no.2 s.62
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• pp.102-111
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• 2006

The increased occurrence of hyperglycemia and oxidative stress in streptozotocin (STZ) induced type I diabetes has been implicated in the etiology and pathology of disease complication. STZ has known to be genotoxic in a variety of assays including tests for microbial mutagenesis and unscheduled DNA synthesis in rat kidney. Diabetes mellitus (DM) is a pathologic condition, resulting in severe metabolic imbalances and non-physiologic changes in many tissues. We examined the effect of gamma radiation and KWNP on preventing the development of insulin dependent diabetes mellitus using streptozotocin-induced Fisher 344 diabetic rats. The hematological values (red blood cell and white blood cell), serum biochemical constituents-alkaline phosphatase (ALP), total cholesterol, triglycerides and insulin-were checked and the organs (testis, spleen and kidney) were weighed. The gonad indices of the STZ treated groups were much lower than the value of the control group. But the gonad indices of the KWNP treated groups were higher than those of the treated groups. The ratio of the weight of kidney to the body weight of the STZ treated groups was higher than that of the control group. The value of the diabetic group treated with KWNP after irradiation (F group) was lower than the other STZ treated groups. The white blood cell and ALP values of the F group were lower than the other STZ groups, as well. The cholesterol and triglyceride values of all the KWNP treated groups were significantly lower than the other groups. A significant increase (about 10 times) of insulin was detected in the F group. The results of hematological assay showed the distinctive damage in the irradiated and STZ treated groups. The quantity of apoptotic cells in seminiferous tubule of testis confirmed a serious damage as assessed in the STZ treated groups. These experimental results have revealed that treatment of the products of KWNP after irradiation has the antidiabetic effect in the STZ-induced diabetic rats. But the F group showed higher recuperative power. These experimental results have revealed that treatment of the gamma irradiation and KWNP have the recovering effect in the STZ-induced diabetic rats.

• Journal of the Korean Applied Science and Technology
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• v.34 no.3
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• pp.568-576
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• 2017

As interest in functionality and environmentally friendly cosmetics is growing in recent years, materials that use safe and effective plant extracts have been developed. Therefore, this study also attempted to check the possibility of the graviola extract, which is known to have various efficacy mainly as a health functional material as a functional cosmetic material. In order to find out the antioxidant activity of graviola, we measured total polyphenol, total flavonoid content and DPPH radical scavenging activity and measured the ROS activity inhibition effect and cytoprotective effect on oxidative stress by treating HDF with hydrogen peroxide cells at an appropriate concentration after checking cytotoxicity in HDF cells. Based on the results of this experiment, the graviola extract was found to contain as high as 26.6 mg(CA)/100g, 14.3 mg(Q)/100g of total polyphenol and flavonoid, which are the antioxidant indexes and to have the high radical scavenging activity. The cell survival rate of the HDF cells was measured, and as a result, no significant cytotoxicity was observed at all concentrations and the experiment was carried out at a concentration of $100{\mu}g/mL$ afterwards. Inhibition of ROS activity in HDF cells induced by hydrogen peroxide was measured and the concentration-dependent inhibition of ROS activity was found and the cell protection effect of graviola was measured after hydrogen peroxide was treated for 4, 24 and 48 hours. As a result, the cell protection effect as high as 89.92% was confirmed at a $25{\mu}g/mL$ concentration up to 24 hours. As these results show that the graviola extract has excellent antioxidant activity, almost no toxicity to HDF cells, an effective activity inhibitory effect on active oxygen generated by hydrogen peroxide and excellent cytoprotective effect, the possibility as various functional materials with antioxidant and cytoprotective effects was confirmed.

• Korean Journal of Clinical Pharmacy
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• v.13 no.1
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• pp.29-39
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• 2003

Neurodegenerative disorders are associated with apoptosis as a causing factor or an inducer. On the other hand, it has been reported that epigallocatechin gallate (EUG), one of antioxidants and flavonoids, and z-VAD-fmk, a nonselective caspase inhibitor, suppress oxidative-radical-stress-induced apoptosis. However, it is not yet known what is the effects of EGCG and z-VAD-fmk on the apoptotic pathway is through phosphoinositide 3-kinase (PI3K), Akt and glycogen synthase kinase-3 (GSK-3) as well as mitochondria, caspase-3 and poly (ADP-ribose) polymerase (PARP). We investigated the effects of EGCG by using $H_2O_2$ treated N18D3 cells, mouse DRG hybrid neurons. Methods: Following 30 min $100\;{\mu}m\;H_2O_2$ exposure, the viability of N18D3 cells (not pretreated vs. EGCG or z-VAD-fmk pretreated) was evaluated by using MTT assay. The effect of EGCG on immunoreactivity (IR) of cytochrome c, caspase-3, PARP, PI3K/Akt and GSK-3 was examined by using Western blot, and was compared with that of z-Y4D-fmk. Results: EGCG or z-VAD-fmk pretreated N18D3 cells showed increased viability. Dose-dependent inhibition of caspase-3 activation accompanied by PARP cleavage were demonstrated by pretreatment of both agents. However, inhibition of cytochrome c release was only detected in EGCG pretreated N18D3 cells. On the pathway through PI3K/Akt and GSK-3, however, the result of Western blot in EGCG pretreated N18D3 cells showed decreased IR of Akt and GSK-3 and increased IR of p85a PI3K, phosphorylated Akt and GSK-3, and contrasted with that in z-VAD-fmk pretreated N18D3 cells showing no changes on each molecule. Conclusion: These data show that EGCG affects apoptotic pathway through upstream signal including PI3K/Akt and GSK-3 pathway as well as downstream signal including cytochrome c and caspase-3 pathway. Therefore, these results suggest that EGCG mediated activation of PI3K/Akt and inhibition GSK-B could be new potential therapeutic strategy for neurodegenerative diseases associated with oxidative injury.

• Journal of the Korean Society of Food Science and Nutrition
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• v.41 no.12
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• pp.1716-1726
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• 2012

This study was undertaken to evaluate the antihyperglycemic, antilipid peroxidative, and antioxidant effects of the ethanol extracts of Artemisia iwayomogi (Ai) in streptozotocin (STZ)-induced diabetic rats. Diabetes was induced in Sprague-Dawley rats with a single intravenous injection (45 mg/kg b.w.) of STZ. The diabetic rats were then randomized to the diabetic and Ai extract therapy groups which were treated with Ai extract at doses of 1, 2, and 3 g/kg b.w./day, respectively, for 14 days. Oral administration of Ai (2 g/kg b.w.) significantly decreased their intake of food. Dosage of 2 g/kg of the extract significantly decreased blood glucose levels in the glucose level in diabetic rats after 4 day, there was no significant difference observed at 1 and 3 g/kg. A dose of 2 or 3 g/kg of the Ai extract significantly reduced plasma glucose levels in STZ-induced hyperglycemic rats at 7 days. The hypoglycemic effect of Ai at a dose of 2 g/kg was significantly more effective than that of STZ-control. The effect was more pronounced in 2 g/kg than 1 g and 3 g/kg. A significant reduction in triglycerides (TG) and free fatty acids (FFA), and a significant increase in liver glycogen were observed in treated diabetic rats at doses of 2 g/kg after 14 days of treatment. Administration of Ai extracts to diabetic rats showed a significant decrease in liver malondialdehyde (MDA) levels. The activity of superoxide dismutase (SOD) was significantly increased in the 3 g extract-supplemented groups. The activities of glutathione peroxidase (GSH-px) and catalase (CAT) were significantly increased in the 1 g and 3 g extract-supplemented groups. Ai extract significantly increased glutathione-S transferase (GST) activity in a dose-dependent manner compared with treatment in STZ-control rats. Our result supports the fact that the administration of Ai extract is able to reduce hyperglycemia and hyperlipidemia risk, and also reduce the oxidative stress in diabetic rats.

• Journal of the Korean Society of Food Science and Nutrition
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• v.41 no.5
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• pp.584-590
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• 2012

Dried $Compositae$ flowers have traditionally been used for the treatment of anti-inflammatory and anti-oxidative stress in Korea. This paper investigates the effects of $Compositae$ extracts on the inhibition of proliferation and apoptosis of human gastric cancer AGS cells, human breast cancer MDA-MB-231 cells, and SK-BR-3 cells. The proliferation of AGS cells, MDA-MB-231 cells, and SK-BR-3 cells were determined by MTT assay. Several $Compositae$ extracts inhibited proliferation of AGS cells, MDA-MB-231 cells, and SK-BR-3 cells in a dose-dependent manner. To assess the apoptosis of $Compositae$ extracts, the nuclei of MDA-MB-231 cells were stained with DAPI. The presence of chromatin condensation in the $Compositae$ extract-treated cells was detected on a fluorescent microscope (${\times}200$). We conducted Western blot analysis of changes in Bcl-2, Bax, and p53 protein expression levels. Apoptosis by $Chrysanthemum$ $zawadskii$ subsp. coreanum, $Chrysanthemum$ $zawadskii$ var. $tenuisectum$ and $Rudbeckia$ $laciniata$ var. $hortensis$ treatment created a decrease in Bcl-2 expression, whereas the expression of Bax and p53 were increased. These results indicate that $Chrysanthemum$ $zawadskii$ $subsp.$ $coreanum$, $Chrysanthemum$ $zawadskii$ var. $tenuisectum$ and $Rudbeckia$ $laciniata$ var. $hortensis$ inhibit breast cancer cell growth through the induction of apoptosis.

• Journal of the Korean Society of Food Science and Nutrition
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• v.41 no.12
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• pp.1663-1670
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• 2012

The inhibitory effects of rose hip (Rosa canina L.) water extracts from two different manufactures on osteoarthritis was comparatively investigated in primary cultures of rat cartilage cells. To identify the effects of rose hip extracts against $H_2O_2$ (300 ${\mu}M$, 2 hr) treatment, cell survival was measured by the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. Cell survival increased by rose hip extracts in the range of 100 to 600 ${\mu}g/mL$ of $H_2O_2$ treatment. To determine the anti-inflammatory effects of rose hip extracts, tumor necrosis factor alpha (TNF-${\alpha}$), nitric oxide (NO), and Cox-2 expression were measured after lipopolysaccharide (LPS) activation. TNF-${\alpha}$ level with rose hip extract treatment was decreased by 27.4% and 31.9% at 600 ${\mu}g/mL$ of $H_2O_2$ treatment. Nitric oxide was inhibited by rose hip extract at 100~600 ${\mu}g/mL$ of $H_2O_2$ treatment in a dose-dependent manner. In addition, Cox-2 protein expression was dose-dependently decreased while Cox-1 had no change in expression level. The severity of osteoarthritis is controlled by a balance between anabolic and catobolic factors in an articulation, therefore the expression of these factors plays a critical role in preventing osteoarthritis. In measuring anabolic factors, the genetic expression of collagen type I increased with rose hip treatment, while the genetic expression of collagen II did not change. In addition, the genetic expression of aggrecan (proteoglycan core protein) was significantly increased. while the genetic expression of matrix metalloproteinase (MMP) 3, 7 and 13, known catabolic factors, was significantly inhibited by treatment with rose hip extract. The expression of MMP13 was especially highly influenced. In conclusion, rose hip water extracts show inhibitory effects on cell death by $H_2O_2$ mediated oxidative stress, which is related to inhibitory effects on inflammation due to TNF-${\alpha}$, NO, and Cox-2. The ability of rose hip extracts to ameliorate inflammation in primary cultures of cartilage cells seems to associate with an increased genetic expression of specific anabolic factors, collagen type I and aggrecan, and a decreased expression of catabolic factors, MMPs (3, 7, and 13). However, there were no significant differences between rose hip extracts from the two manufacturers.