• Molecules and Cells
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• v.21 no.2
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• pp.206-212
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• 2006

We have established in culture a spontaneously immortalized bovine embryonic fibroblast (BEF) cell line that has lost p53 and $p16^{INK4a}$ functions. MyoD is a muscle-specific regulator capable of inducing myogenesis in a number of cell types. When the BEF cells were transduced with MyoD they differentiated efficiently to desmin-positive myofibers in the presence of 2% horse serum and 1.7 nM insulin. The myogenic differentiation of this cell line was more rapid and obvious than that of C2C12 cells, as judged by morphological changes and expression of various muscle regulatory factors. To confirm that lack of the p53 and $p16^{INK4a}$ pathway does not prevent MyoD-mediated myogenesis, we established a cell line transformed with SV40LT (BEFV) and introduced MyoD into it. In the presence of 2% horse serum and 1.7 nM insulin, the MyoD-transduced BEFV cells differentiated like the MyoD-transduced BEFS cells, and displayed a similar pattern of expression of muscle regulatory proteins. Taken together, our results indicate that MyoD overexpression overcomes the defect in muscle differentiation associated with immortalization and cell transformation caused by the loss of p53 and Rb functions.

• Journal of Life Science
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• v.31 no.11
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• pp.969-979
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• 2021

Type 2 diabetes is a serious chronic metabolic disease, and the goal of diabetes treatment is to keep blood glucose at a normal level and prevent complications from diabetes. Hyperglycemia is a key pathologic feature of type 2 diabetes that mainly results from insulin resistance and pancreatic β-cell dysfunction. Chronic exposure of β-cells to elevated glucose concentrations induces glucotoxicity. In this study, we examined whether an 80% ethanol extract of Oxya chinensis sinuosa Mishchenko (OEE) protected INS-1 pancreatic β-cells against glucotoxicity-induced apoptosis and oxidative stress. Pretreatment with a high concentration of glucose (high glucose = 30 mM) induced glucotoxicity and apoptosis of INS-1 pancreatic β cells. Treatment with OEE significantly increased cell viability. Treatment with 0.01-0.20 mg/ml OEE dose dependently decreased intracellular reactive oxygen species, lipid peroxidation, and nitric oxide levels and increased insulin secretion in high glucose-pretreated INS-1 β cells. OEE also significantly increased the activities of antioxidant enzymes in response to high-glucose-induced oxidative stress. Moreover, OEE treatment significantly reduced the expressions of pro-apoptotic proteins, including Bax, cytochrome C, caspase-3, and caspase-9, and increased anti-apoptotic Bcl-2 expression. Apoptotic cells were identified using Annexin-V/propidium iodide staining, which revealed that treatment with OEE significantly reduced high-glucose-induced apoptosis. These findings implicate OEE as a valuable functional food in protecting pancreatic β-cells against glucotoxicity-induced apoptosis and oxidative stress.

• Asian-Australasian Journal of Animal Sciences
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• v.31 no.1
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• pp.138-148
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• 2018

Objective: Fusarium mycotoxins deoxynivalenol (DON) and zearalenone (ZEN), common contaminants in the feed of farm animals, cause immune function impairment and organ inflammation. Consequently, the main objective of this study was to elucidate DON and ZEN effects on the mRNA expression of pro-inflammatory cytokines and other immune related genes in the kidneys of piglets. Methods: Fifteen 6-week-old piglets were randomly assigned to three dietary treatments for 4 weeks: control diet, and diets contaminated with either 8 mg DON/kg feed or 0.8 mg ZEN/kg feed. Kidney samples were collected after treatment, and RNA-seq was used to investigate the effects on immune-related genes and gene networks. Results: A total of 186 differentially expressed genes (DEGs) were screened (120 upregulated and 66 downregulated). Gene ontology analysis revealed that the immune response, and cellular and metabolic processes were significantly controlled by these DEGs. The inflammatory stimulation might be an effect of the following enriched Kyoto encyclopedia of genes and genomes pathway analysis found related to immune and disease responses: cytokine-cytokine receptor interaction, chemokine signaling pathway, toll-like receptor signaling pathway, systemic lupus erythematosus (SLE), tuberculosis, Epstein-Barr virus infection, and chemical carcinogenesis. The effects of DON and ZEN on genome-wide expression were assessed, and it was found that the DEGs associated with inflammatory cytokines (interleukin 10 receptor, beta, chemokine [C-X-C motif] ligand 9, CXCL10, chemokine [C-C motif] ligand 4), proliferation (insulin like growth factor binding protein 4, IgG heavy chain, receptor-type tyrosine-protein phosphatase C, cytochrome P450 1A1, ATP-binding cassette sub-family 8), and other immune response networks (lysozyme, complement component 4 binding protein alpha, oligoadenylate synthetase 2, signaling lymphocytic activation molecule-9, ${\alpha}$-aminoadipic semialdehyde dehydrogenase, Ig lambda chain c region, pyruvate dehydrogenase kinase, isozyme 4, carboxylesterase 1), were suppressed by DON and ZEN. Conclusion: In summary, our results indicate that high concentrations of DON and ZEN suppress the inflammatory response in kidneys, leading to potential effects on immune homeostasis.

• Journal of Life Science
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• v.31 no.9
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• pp.796-805
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• 2021

Hepatic endoplasmic reticulum (ER) stress contributes to the development of steatosis and insulin resistance. The components of unfolded protein response (UPR) regulate lipid metabolism. Recent studies have reported an association between ER stress and aberrant cellular lipid control; moreover, research has confirmed the involvement of sterol regulatory element-binding proteins (SREBPs)-the central regulators of lipid metabolism-in the process. However, the exact role of SREBPs in controlling lipid metabolism during ER stress and its contribution to fatty liver disease remain unknown. Here, we show that SREBP-1c deficiency protects against ER stress-induced hepatic steatosis in mice by regulating UPR, inflammation, and fatty acid oxidation. SREBP-1c directly regulated inositol-requiring kinase 1α (IRE1α) expression and mediated ER stress-induced tumor necrosis factor-α activation, leading to a reduction in expression of peroxisome proliferator-activated receptor γ coactivator 1-α and subsequent impairment of fatty acid oxidation. However, the genetic ablation of SREBP-1c prevented these events, alleviating hepatic inflammation and steatosis. Although the mechanism by which SREBP-1c deficiency prevents ER stress-induced inflammatory signaling remains to be elucidated, alteration of the IRE1α signal in SREBP-1c-depleted Kupffer cells might be involved in the signaling. Overall, the results suggest that SREBP-1c plays a crucial role in the regulation of UPR and inflammation in ER stress-induced hepatic steatosis.

• Toxicological Research
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• v.24 no.3
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• pp.175-182
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• 2008

DNA-dependent protein kinase(DNA-PK) is involved in joining DNA double-strand breaks induced by ionizing radiation or V(D)J recombination and is activated by DNA ends and composed of a DNA binding subunit, Ku, and a catalytic subunit, DNA-PKcs. It has been suggested that DNA-PK might be $2^{nd}$ upstream kinase for protein kinase B(PKB). In this report, we showed that Ser473 phosphorylation in the hydrophobic-motif of PKB is blocked in DNA-PK knockout mouse embryonic fibroblast cells(MEFs) following insulin stimulation, while there is no effect on Ser473 phosphorylation in DNA-PK wild type MEF cells. The observation is further confirmed in human glioblastoma cells expressing a mutant form of DNA-PK(M059J) and a wild-type of DNA-PK(M059K), indicating that DNA-PK is indeed important for PKB activation. Furthermore, the treatment of cells with doxorubicin, DNA-damage inducing agent, leads to PKB phosphorylation on Ser473 in control MEF cells while there is no response in DNA-PK knockout MEF cells. Together, these results proposed that DNA-PK has a potential role in insulin signaling as well as DNA-repair signaling pathway.

• Journal of Life Science
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• v.18 no.1
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• pp.1-8
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• 2008

The purpose of this study was to determine the possible additive effects of endurance exercise training (EXER) and selenium (SELE) on the improvements of glucose and lactate transport capacities in diabetic Goto-kakizaki rats. Animals either remained sedentary control (SED) or performed EXER or received SELE [$5{\mu}mol$ kg body wt (-1) day (-1)], or underwent both EXER and SELE (COMBI), which lasted for 6 wk. Compared with sedentary control, EXER alone or the SELE alone group, or the combined treatment group had significant reduction in glucose response measured at 90 min and 120 min during an intraperitoneal glucose tolerance test (IPGTT) and body weight after 6week treatment. EXER alone, or combined group individually had significantly higher glycogen contents in liver compared with SED or SELE groups. EXER alone increased glycogen content in soleus and plantaris compared with SED, and this parameter was increased to greatest extent in the combined treatment groups compared with SED or SELE groups. EXER alone, SELE alone or COMBI, caused significant decreases in the plasma lactates, serum glucose, insulin, total cholesterol and HOMA-IR along with a significant increase in high-density lipoprotein cholesterol compared with SED. In addition, EXER or COMBI individually had significantly lower serum triacylglycerol compared with SED or SELE. With respect to protein expression related to glucose and lactate transport capacities, EXER alone, SELE alone, or COMBI increased in MCT1 and MCT4 protein level in soleus and plantaris. Furthermore, EXER alone, SELE alone or COMBI caused significant increases in mt MCT1 protein level in soleus and plantaris. The findings of the current study suggest that endurance exercise training and selenium treatment may provide therapeutic values to type II diabetic patients with peripheral insulin resistance and hyperlactatecemia by improving glucose and lactate transport capacities, leading to improvements in plasma lactate, serum glucose, insulin and lipid profiles (TC, TG, HDL).

• Food Science of Animal Resources
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• v.24 no.3
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• pp.283-292
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• 2004

This study was conducted to investigate feeding effects of the high pressure boiled extracts (HPBE) of the Ogol chicken with herbs on glucose, hormones and immunological response (cytokine, specific antibody) of serum in the rat which fed either with normal feed (T$_1$), normal feed + herb HPBE (T$_2$), normal feed + Ogol chicken HPBE (T$_3$), normal feed + mixture of cross-bred Ogol chicken HPBE (T$_4$) hydrolyzed with Flavourzyme 0.1% for 35 days. During experimental period, there was a weak trend to have a higher glucose content for the T$_4$ group with 102.27${\pm}$5.95 mg/dL, but it was not significantly higher than other treatments. For insulin level, T$_1$ group showed numerically a slightly higher level with 6.79${\pm}$4.64 ${\mu}$IU/mL, but the difference was not significant in statistic term due likely to a large variation in comparison with other treatments. The treatments did not significantly alter testosterone level in rat plasma with 1.09, 1.46, 0.98, 1.13 ng/mL in T$_1$, T$_2$, T$_3$ and T$_4$, respectively. T$_4$ treatment increased the aldosterone level to a significantly (p<0.05) higher level (273.33 ng/dL) than other treatments. The extract treated rat showed a tendency in the cortisol level of lower levels than the control group, particularly, it was significantly (p<0.05) lower in T$_3$ group than other groups. T$_3$ and T$_4$ groups showed higher levels for interlukin-4 (IL-4) and anti-BSA IgG in immune cells and plasma. T$_2$, T$_3$ and T$_4$ treatments showed a slightly higher levels in v-interferon (INF-r) than the control, with a greater effect for T4 treatments. These results suggested that HPBE of the cross-bred Ogol chicken hydrolyzed with Flavourzyme increased immunological activity and decreased the concentration of cortisol and aldosterone hormones.

• Journal of the Korean Society of Food Science and Nutrition
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• v.34 no.8
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• pp.1202-1209
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• 2005

The purpose of this study was to observe the effects of high-carbohydrate (HC) diet, high-protein (HP) diet and high-fat (HF) diet with a normal diet (N) on diet-induced thermogenesis (DIT) in healthy Korean women. The four isoenergetic test meals consisted as follows: HC (75$ \% $ energy from carbohydrate, 10$ \% $ from protein and 15$ \% $ from fat), HP (10$ \% $ energy from carbohydrate, 75$ \% $ from protein and 15$ \% $ from fat) HF (15$ \% $ energy from carbohydrate, 10$ \% $ from protein and 75$ \% $ from fat) diet and N (65$ \% $ energy from carbohydrate, 15$ \% $ from protein and 20$ \% $ from fat) diet. Fasting and postprandial thermogenesis were measured after each test meals in eight subjects by indirect calorimetry for 3 hours. Fasting and postprandial serum glucose, insulin and triacylglycerol contents were also measured for 3 hours. DIT were 10.4$\pm$3.2 for HC, 12.7$\pm$0.5 for HP, 6.9$\pm$2.8 for HF and 8.7$\pm$4.8 for N diet as a percentage of the energy load. DIT of HP was greater than HF significantly, but had no differences with HC and N Integrated areas under the curves ($ \delta- $-AUC) of plasma glucose and insulin were significantly higher for HC compared to HP, HF and N diets. $ \delta$-AUC of plasma triacylglycerol was significantly higher for HF compared to the other test meals. In conclusion, intake of protein rich meal stimulates postprandial energy expenditure, but has no relation between DIT and insulin response, body composition.

• Food Science and Biotechnology
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• v.17 no.1
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• pp.130-134
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• 2008

Hypoglycemic effect of onion skin extract in vitro and in vivo was investigated. A methanol extract of onion skin inhibited yeast $\alpha$-glucosidase with an $IC_{50}$ of 0.159 mg/mL. A single oral administration of the onion skin extract (500 mg/kg) significantly lowered the postprandial area under the glucose response curve to starch (1 g/kg, p<0.05). Three-week-old db/db mice were fed an AIN-93G diet or a diet supplemented with a 0.5% onion skin extract for 7 weeks after a 1-week adaptation period. Consumption of onion skin extract significantly reduced the levels of plasma glucose, insulin, and blood glycated hemoglobin as compared with the control group (p<0.05). These findings suggest that onion skin is effective in controlling hyperglycemia in animal models of type 2 diabetes mellitus, at least in part by inhibiting $\alpha$-glucosidase activity.

• Korean Journal of Animal Reproduction
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• v.21 no.4
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• pp.325-333
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• 1997

Recent evidence indicates that growth factors modulate response of mammary epithelial cells to environmental stress. The objective of this study was to examine the cellular and biochemical responses of mammary tissue to environmental stress caused by artificial mastitis. For experimental a, pp.oach, toxins of most mastitis causing organisms(Staph. aureus or Strep. agalactiae) and heat stress(42$^{\circ}C$) were artificially exposed to mammary tissue. Effects of these environmental stresses on cell growth, cell death and heat shock protein synthesis were examined. Lactating mammary tissure were cultured under basal medium(DMEM) su, pp.emented with insulin(10$\mu\textrm{g}$/ml) and aldosterone(1$\mu\textrm{g}$/ml). All treatment groups in heat stress at 42$^{\circ}C$ incubation significantly decreased DNA synthesis rates in comparison with those at 39$^{\circ}C$(P<0.05), however, these decreased DNAa synthesis rates were recovered by addition of adenosine(10$\mu$M) and IGFI(10ng/ml). Similar results were obtained when tissue growth rates were measured by DNA content/tissue. Strep. agalactiae toxin did not significantly decreased DNA content/tissue in comparison with no treatment of bacterial toxin with or without heat stress, however, tended to decrease DNA contents/tissue without heat stress. In the fluorography analysis, heat stress(42$^{\circ}C$ incubation) slightly increased 35S-methoionine labelled 70kd protein synthesis. These results indicate that environmental stress caused by artificial mastitis slightly decreased mammary growth or mammary size, however, these results could be recovered by addition of adenosine and IGF-I.