• Korean Journal of Poultry Science
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• v.44 no.1
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• pp.1-9
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• 2017

The objective of the present study was to determine the expression of genes associated with lipopolysaccharide (LPS)-induced stressor in two breeds of chickens: the Korean native chicken (KNC) and the White Leghorn chicken (WLH). Forty chickens per breed, aged 40 weeks, were randomly allotted to the control (CON, administered the saline vehicle) and LPS-injected stress groups. Samples were collected at 0 and 48 h post-LPS injection, and total RNA was extracted from the chicken livers for RNA microarray and quantitative real-time polymerase chain reaction (qRT-PCR) analyses. In response to LPS, 1,044 and 1,193 genes were upregulated, and 1,000 and 1,072 genes were downregulated in the KNC and WLH, respectively, using a ${\geq}2$-fold cutoff change. A functional network analysis revealed that stress-related genes were downregulated in both KNC and WLH after LPS infection. The results obtained from the qRT-PCR analysis of mRNA expression of heat shock 90 (HSP90), 3-hydroxy-3-methylglutaryl-CoA reductase (HMGCR), activating transcription factor 4 (ATF4), sterol regulatory element-binding protein 1 (SREBP1), and X-box binding protein 1 (XBP1) were confirmed by the results of the microarray analysis. There was a significant difference in the expression of stress-associated genes between the control and LPS-injected KNC and WLH groups. The qRT-PCR analysis revealed that the stress-related $HSP90{\alpha}$ and HMGCR genes were downregulated in both LPS-injected KNC and WLH groups. However, the HSP70 and $HSP90{\beta}$ genes were upregulated only in the LPS-injected KNC group. The results suggest that the mRNA expression of stress-related genes is differentially affected by LPS stimulation, and some of the responses varied with the chicken breed. A better understanding of the LPS-induced infective stressors in chicken using the qRT-PCR and RNA microarray analyses may contribute to improving animal welfare and husbandry practices.

• Journal of the Korean Applied Science and Technology
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• v.34 no.3
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• pp.613-622
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• 2017

The object of this study was to determine the influence of dietary metabolic energy (ME) on ..... A total of 240 meat ducks Cherry valley (Anas platyrhynchos) were assigned into four treatment groups with a randomized block design for 42 days. The four treatments were: ME 2900 kcal/kg, ME 3000 kcal/kg, ME 3100 kcal/kg, and ME 3200 kcal/kg. There was no difference in liver tissue among the treatments. The duodenal villi and crypt depth length decreased by 10.58% in 2900 compared with ME 3000, but there was no difference between 3100 and 3200. Counts of caecal Latobacillus decreased by 9.47% in 2900 compared to ME 3000, but increased by 2.52 and 3.24% in 3100 and 3200, respectively. Total aerobic bacteria, E. coli and Coliform bacteria were increased by 2900 when compared to ME 3000, but there was no difference between 3100 and 3200. HSP $90-{\alpha}$ among the heat shock proteins (HSPs)-mRNA in the liver was reduced by 48.60% in 2900 compared to ME 3000, while 3100 and 3200 showed no difference or increased.

• Korean Journal of Poultry Science
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• v.40 no.2
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• pp.147-155
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• 2013

This study was conducted to evaluate the effects of dietary supplementation of vitamin C or E on the expressions of endoplasmic reticulum (ER) stress, lipid and glucose metabolism associated genes in broiler chickens. A total of 216 one-day-old male broilers was randomly alloted to 4 treatments with 6 replicate pens per treatment and 9 broilers per pen for 35 days. The dietary treatments were control, vitamin C (control diet + ascorbic acid 200 mg/kg diet), vitamin E (control diet + ${\alpha}$-tocopherol 100 mg/kg diet), vitamin C + E (control diet + vitamin C 200 mg/kg + vitamin E 100 mg/kg), respectively. To evaluate gene expressions by quantitative real-time polymerase chain reaction, total RNA was extracted from the liver of the chicken at 35 days of age. Dietary supplementation of vitamins was significantly down-regulated the expression of stress marker genes including HSP70, HSP90, and HMGCR, as compared to the control (p<0.05). The expressions of ER stress associated genes also inhibited by supplementation of vitamins as well (p<0.05). Vitamin C supplementation suppressed the expression of lipid associated genes such as FASN, FATP1 and ACSL1. Vitamin supplementation did not affect the glucose transporters, GLUT2 and GLUT8, in the liver. The results of the present study indicated that dietary supplementation of vitamin C or E could be beneficial for the alleviating physiological stress in broiler chickens.

• Journal of Pharmacopuncture
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• v.12 no.1
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• pp.5-12
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• 2009

Background : Anticancer effects of herbal medicine have been reported in various types of cancer, but the systematic approaches to explain molecular mechanism(s) are not established yet. Objective : To find the anticancer-effect and mechanism(s) of Water Extract of Coptis japonica (WECJ) colon cancer cell (SNU-81). Methods : We first selected 11 herbals, and anti-cancer effects of water-extracts from those herbals have been tested in human colon cancer cell line, SNU-81. Among the tested herbals, the WECJ significantly reduced proliferation of SNU-81. To establish a basis of understanding for anti-cancer mechanism, whole proteins have been obtained from SNU-81 harvested at 48 and 96 hrs after the treatment of WECJ, protein expression has been profiled by 2DE-based proteomic approach. Results : Various changes of the protein expression have been monitored, and most frequent dysregulation was found in the molecular chaperons including heat shock protein 90-alpha (Hsp90-alpha), 14-3-3 protein epsilon, T-complex protein 1 subunit alpha, protein disulfide-isomerase A3, and calreticulin. Interestingly, proliferation-associated protein 2G4 has been up-regulated, and it suggests the possible effect of Coptis japonica on ErbB3-regulated signal transduction pathway and growth control of human colon cancer cells. Conclusion : Based upon the present findings, the further study will focus on monitoring various cancer survival factors after artificial regulation of the proteins identified, and it would be the basis for the understanding of the Coptis japonica anti-cancer effect(s) at the molecular level.

• YAKHAK HOEJI
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• v.50 no.4
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• pp.263-267
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• 2006

CD1d is an unique antigen presenting molecule which provides antigenic repertoires to NKT cells. To examine molecules required for CD1d antigen presentation, we determined an interaction between CD1d and several endoplasmic reticulum (ER) resident molecular chaperones by co-immunoprecipitation. Results indicated that calnexin and calreticulin seem to be bound to mouse CD1d, but TAP and tapasin do not bind. Further, we screened an yeat two hybrid system to identify proteins that help mouse CD1d transportation in the cytosol. We found that two proteins, heat shock protein a sub-unit $(Hsp90{\alpha})$ and protein kinase C and casein kinase substrate in neurons 3 (PACSIN-3), interact with CD1d. Future study will be focus on the role of these molecules during the CD1d antigen presentation.

• Korean Journal of Poultry Science
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• v.42 no.1
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• pp.51-59
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• 2015

Chickens are exposed to the external and internal stressors such as low and high temperature, high stocking density, feed restriction and disease. There have been a few studies on gene expressions through the investigation of chickens under direct exposure to the stress of high stocking density. The objective of the present study was to determine the expressions of genes associated with stress, endoplasmic reticulum (ER)-stress, lipid and glucose metabolism in two strains of chickens, Korean Native Chicken (KNC) and White Leghorn (WL), raised in high stocking density. A total of 164 chickens aged 40 weeks were randomly allotted to a $540cm^2/bird$ stocking density (control), whereas the chickens in a high density group were assigned in a $311cm^2/bird$ stocking density with feeding ad libitum for 10 weeks. Total RNA was extracted from the live for qRT-PCR. The expression levels of hsp70 and $hsp90{\alpha}$ were higher in WL subjected to stress with high stocking density compared with those genes in control (P<0.05), while the expressions of genes were not affected in KNC. ER stress marker gene XBP1 was also highly expressed in WL with stress (P<0.05), but the stress of high stocking density did not influence to ER stress marker genes in KNC. Lipid metabolism associated genes including FABP4, FATP1 and ACSL1 were highly expressed in WL compared with KNC when subjected to high stocking density stress (P<0.05). The expression of glucose transport gene GLUT2 and GLUT8 were increased in chickens exposured to the stress of high stocking density (P<0.05). The data indicate that WL is more sensitive to the stress of high stocking density compared with KNC and the stress may influence the modulation of lipid and glucose metabolism in the liver of chickens.

• Korean Journal of Poultry Science
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• v.43 no.3
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• pp.177-189
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• 2016

This study was conducted to verify the relationships between the expression values of stress-related markers and their production performances in 25 strains of Korean domestic chicken breeds. For stress response markers, the amount of telomeric DNA; expression levels of heat shock protein (HSP)-70, $HSP-90{\alpha}$, and $HSP-90{\beta}$; and comet scores were analyzed. Production performances were measured by the survival rate, body weights, days at first egg laying, egg weight and hen housed egg production. The results showed that the production traits and values of stress-related markers showed significant differences between strains. In general, the stress response of pure bred chickens with heavy weights was relatively high, while that of hybrid chickens with light weights was relatively low. The correlation coefficients between telomere contents and body weights showed that there were weak negative relationships. However, the correlations of telomere content with the survival rate and egg production were weakly positive after 20 weeks old. The expression levels of HSP genes and DNA damage rate (comet scores) were positively correlated to body weight, but were negatively correlated to the survival rate and egg production. The results implied that increasing body weight was associated with increasing HSPs expression and the DNA damage rate was associated with decreasing telomere content. In addition, increasing HSPs expression and the DNA damage rate decreased the survival rate and egg production, but the relationships with the telomere content was the reverse. Correlations among the stress-related markers showed that there were significant correlation coefficients between all of the marker values. HSPs expression was negatively correlated to the telomere content, while it was positively correlated to the DNA damage rate. There was a highly negative correlation between the telomere content and DNA damage rate. In conclusion, increasing the HSP values and DNA damage rate can promote telomere reduction, which led to a decrease in disease resistance and robustness of the chicken. Thus, increasing the stress response was verified to adversely affect the laying performance and viability of chickens.

• Journal of Microbiology and Biotechnology
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• v.33 no.10
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• pp.1390-1401
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• 2023

In this study, a 12-week feeding experiment was conducted to characterize the effects of exogenous α-amylase on the growth, feed utilization, digestibility, plasma α-amylase activity, feed degradation rate, and fecal particle size of olive flounder (Paralichthys olivaceus). Diet was supplemented with 0 (AA₀; control), 100 (AA₁₀₀), 200 (AA₂₀₀), or 400 (AA₄₀₀) mg/kg of α-amylase, respectively. Fish (273.1 ± 2.3 g) were stocked into 12 tanks (25 fish/1,000-L tank) and 3 tanks were randomly selected for each diet group. As a result, α-amylase was found to have no significant effects (p ≥ 0.05) on the growth, feed utilization parameters, and whole-body proximate compositions. α-Amylase-treated fish exhibited only a significant increase in the apparent digestibility coefficient of carbohydrates compared to the controls. In addition, in vitro analyses revealed that α-amylase dose-dependently increased (p < 0.05) the feed degradation rate, while photographs of the intestinal content after 2, 4, and 8 h of feeding demonstrated an improved degradation rate in the α-amylase-treated groups. Plasma α-amylase content was higher in the AA₂₀₀ and AA₄₀₀ groups, whereas the control group produced significantly larger-sized fecal particles (90% size class) than these two groups. In the intestine, no changes were observed in the expression levels of the immune-related TNF-α, IL-1β, IL-2, immunoglobulin-M, HSP-70, lysozyme, and amylase alpha-2A. However, growth-related genes IGF-1, IGF-2, TGF-β3, and growth hormone genes were upregulated in muscle tissues. Collectively, exogenous α-amylase has positive roles in the modulation of the digestibility coefficient, blood α-amylase concentration, growth-related gene expression, and diet degradation for improved digestion in olive flounder.

• Journal of Life Science
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• v.22 no.12
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• pp.1672-1679
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• 2012

The effect of high stocking density (HSD) on the expression of stress and lipid metabolism associated genes in the liver of broiler chickens was examined by chicken genome array analysis. The chickens in a control group were randomly assigned to a $495cm^2/bird$ stocking density, whereas the chickens in a HSD group were arranged in a $245cm^2/bird$ stocking density with feeding ad libitum for 35 days. The chickens assigned to the HSD group had a significantly lower body weight, weight gain, and feed intake compared with those of the control group (p<0.05). The mortality of chickens was higher in the HSD group than in the control group. The microarray analysis indicated up-regulation of stress associated genes such as HMGCR, $HSP90{\alpha}$, HSPA5 (GRP78/Bip), DNAJC3 and ATF4, and down-regulation of interferon-${\gamma}$ and PDCD4 genes. The endoplasmic reticulum stress associated genes, HSPA5 (GRP78/Bip), DNAJC3 and ATF4, were highly expressed in the HSD group. The genes, ACSL5, TMEM195 and ELOVL6, involved in fatty acid synthesis, were elevated in the HSD group. The genes, ACAA1, ACOX1, EHHADH, LOC423347 and CPT1A, related to fatty acid oxidation, were also activated in the HSD group. These results suggest that a HSD rearing system stimulates the genes associated with fatty acid synthesis as well as fatty acid oxidation in the liver of broiler chickens.

• The Journal of Internal Korean Medicine
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• v.29 no.2
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• pp.469-489
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• 2008

Objectives : The purpose of this study was to investigate the effects of Chungganhaeju-tang(Qingganjiejiu-tang) on alcoholic liver damaged by applying proteomics. Materials and Methods : Sprague-Dawley rats were used in this experiment the rats were divided into the normal group, the control group(alcohol) and the sample group(CGHJT +alcohol). The ethanol was orally administered twice a day for 6 weeks in the control and sample groups. Water instead of ethanol was orally administered twice a day for 6 weeks in the normal group. CGHJT extract was orally administered once a day for 6 weeks in the sample group. The livers of each group were processed and assessed by histology, Western Blot, $Oxyblot^{TM}$, CBB and 2-dimensional electrophoresis. Results : In the histological findings of the liver, CGHJT inhibited hepatic fibrogenesis induced by alcohol. TIMP-1 decreased in the sample group assessed by western blot and statistical significance was noted by dot blotting(p<0.05). In the $Oxyblot^{TM}$, protein oxidation induced by alcohol treatment decreased with CGHJT. In the 2-dimensional electrophoresis finding, increased proteins alcohol such as HSP 60, 60kDa heat shock protein, 3-mercaptopyruvate sulfurtransferase were normalized by CGHJT. CGHJT was considered to normalize the anti-oxidation activity elevated by alcohol. In the 2-dimensional electrophoresis finding, increased oxidized proteins such as actin, prolyl 4-hydroxylase beta polypeptide, 94kDa glucose regulated protein(GRP94), heat shock protein 90-alpha(HSC86), calreticulin precursor(CRP55), ATP synthase beta chain mitochondrial precursor, caspase-8 precursor, and dihydrolipoamide succinyltransferase(E2) decreased with CGHJT. CGHJT was considered to reduce the oxidative stress of alcohol. Conclusion : Chungganhaeju-tang(Qingganjiejiu-tang) exerts an inhibitory effect against the fibrosis and protein oxidation induced by alcohol treatment of rat liver. CGHJT was considered to normalize the elevated anti-oxidation activity by alcohol and to reduce the level of oxidative stress due to alcohol.