• Journal of Veterinary Science
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• 제23권6호
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• pp.90.01-90.13
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• 2022

Background: Insulin regulates glucose homeostasis and has important effects on metabolism, cell growth, and differentiation. Depending on the cell type and physiological context, insulin signal has specific pathways and biological outcomes in different tissues and cells. For studying the signal pathway of insulin on glycolipid metabolism in porcine embryonic fibroblast (PEF), we used high-throughput sequencing to monitor gene expression patterns regulated by insulin. Objectives: The goal of our research was to see how insulin affected glucose and lipid metabolism in PEFs. Methods: We cultured the PEFs with the addition of insulin and sampled them at 0, 48, and 72 h for RNA-Seq analysis in triplicate for each time point. Results: At 48 and 72 h, 801 and 1,176 genes were differentially expressed, respectively. Of these, 272 up-regulated genes and 264 down-regulated genes were common to both time points. Gene Ontology analysis was used to annotate the functions of the differentially expressed genes (DEGs), the biological processes related to lipid metabolism and cell cycle were dominant. And the DEGs were significantly enriched in interleukin-17 signaling pathway, phosphatidylinositol-3-kinase-protein kinase B signaling pathway, pyruvate metabolism, and others pathways related to lipid metabolism by Kyoto Encyclopedia of Genes and Genomes enrichment analysis. Conclusions: These results elucidate the transcriptomic response to insulin in PEF. The genes and pathways involved in the transcriptome mechanisms provide useful information for further research into the complicated molecular processes of insulin in PEF.

• Journal of Animal Science and Technology
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• 제64권3호
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• pp.481-499
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• 2022

This study aims to determine the effects of D-methionine (D-Met) isomer and the methionine precursor 2-hydroxy-4-methylthiobutanoic acid i (HMBi) supplementation on milk protein synthesis on immortalized bovine mammary epithelial cell (MAC-T). MAC-T cells were seeded using 10-cm dishes and cultured in Dulbecco's modified Eagle's medium/F12 (DMEM/F12) basic medium. The basic medium of DMEM/F12 was replaced with the lactogenic DMEM/ F12 differentiation medium when 90% of MAC-T cells reached confluency. The best dosage at 0.6 mM of D-Met and HMBi and incubation time at 72 h were used uniformly for all treatments. Each treatment was replicated six times wherein treatments were randomly assigned in a 6-well plate. Cell, medium, and total protein were determined using a bicinchoninic acid protein assay kit. Genes, proteomics and metabolomics analyses were also done to determine the mechanism of the milk protein synthesis pathway. Data were analyzed by two-way analysis of variance (ANOVA) with supplement type and plate as fixed effects. The least significant difference test was used to evaluate the differences among treatments. The HMBi treatment group had the highest beta-casein and S6 kinase beta-1 (S6K1) mRNA gene expression levels. HMBi and D-Met treatments have higher gene expressions compared to the control group. In terms of medium protein content, HMBi had a higher medium protein quantity than the control although not significantly different from the D-Met group. HMBi supplementation stimulated the production of eukaryotic translation initiation factor 3 subunit protein essential for protein translation initiation resulting in higher medium protein synthesis in the HMBi group than in the control group. The protein pathway analysis results showed that the D-Met group stimulated fructose-galactose metabolism, glycolysis pathway, phosphoinositide 3 kinase, and pyruvate metabolism. The HMBi group stimulated the pentose phosphate and glycolysis pathways. Metabolite analysis revealed that the D-Met treatment group increased seven metabolites and decreased uridine monophosphate (UMP) production. HMBi supplementation increased the production of three metabolites and decreased UMP and N-acetyl-L-glutamate production. Taken together, D-Met and HMBi supplementation are effective in stimulating milk protein synthesis in MAC-T cells by genes, proteins, and metabolites stimulation linked to milk protein synthesis.

• Molecules and Cells
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• 제24권3호
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• pp.394-408
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• 2007

Several genes/QTLs governing resistance/tolerance to abiotic and biotic stresses have been reported and mapped in rice. A QTL for submergence tolerance was found to be co-located with a major QTL for broad-spectrum bacterial leaf blight (bs-blb) resistance on the long arm of chromosome 5 in indica cultivars FR13A and IET8585. Using the Nipponbare (japonica) and 93-11 (indica) genome sequences, we identified, in silico, candidate genes in the chromosomal region [Kottapalli et al. (2006)]. Transcriptional profiling of FR13A and IET8585 using a rice 22K oligo array validated the above findings. Based on in silico analysis and arraying we observed that both cultivars respond to the above stresses through a common signaling system involving protein kinases, adenosine mono phosphate kinase, leucine rich repeat, PDZ/DHR/GLGF, and response regulator receiver protein. The combined approaches suggest that transcription factor EREBP on long arm of chromosome 5 regulates both submergence tolerance and blb resistance. Pyruvate decarboxylase and alcohol dehydrogenase, co-located in the same region, are candidate downstream genes for submergence tolerance at the seedling stage, and t-snare for bs-blb resistance. We also detected up-regulation of novel defense/stress-related genes including those encoding fumaryl aceto acetate (FAA) hydrolase, scramblase, and galactose oxidase, in response to the imposed stresses.

• Journal of Ginseng Research
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• 제17권2호
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• pp.101-108
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• 1993

This study was made to understand a hypoglycemic action of the fat soluble fraction of Panax ginseng C.A. Meyer in streptozotocin induced diabetic rats by determining the activities of several enzymes related to carbohydrate and lipid metabolism as well as several blood component levels such as glucose and ketone bodies, and non-esterified fatty acids. Albino rats (Sprague Dawley, 170-200g, 3) were injected once with 70mg streptozotocinhg body weight intraperitoneally and fed with ordinary diet for 7 days, and then the fat soluble fraction (5 mg~20 mg/day/rat) was injected intraperitoneally once a day for three days to rats having high blood glucose level over 340 mg/100ml. After a final injection of the fat soluble fraction, rats u.ere starved for 16 hours followed by the analysis of blood serum and liver enzymes. It was found that increased levels of glucose, ketone bodies and free fatty acids in streptozotocin induced rats were decreased appreciably by administration of the fat soluble fraction. However, the amount of administered fat soluble fraction did not show any significantly different hypoglycemic action. Decreased activities of glucokinase, phosphofructokinase, pyruvate kinase, 6-phosphogluconate dehydrogenase and acetyl CoA carboxylase of the liver of streptozotocin induced diabetic rats were greatly modified suggesting that a hypoglycemic action of the fat soluble fraction was also appreciable as ginseng saponin fraction. We also compared a hypoglycemic action of the fat soluble fraction prepared from American ginseng and Chinese ginseng with that of Korean pain ginseng. 핀o significant difference of the hypoglycemic activity was observed between the above ginseng fat soluble fractions, suggesting that a study of the fat soluble fraction might be one of the most interesting subjects relating to diabetic hyperglycemia in the near future.

• Asian-Australasian Journal of Animal Sciences
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• 제15권8호
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• pp.1215-1221
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• 2002

Normal haematological and biochemical indices along with thyroid hormone status were studied in healthy Kathiawari horses of different age groups (yearling, young stock, adults and old stock) belonging to either sex. Effect of both age and sex was observed on thyroid hormone levels, haematological and biochemical indices. In females, hemoglobin levels was significantly lower in yearlings than adult animals while total leukocyte counts were higher in yearlings than equids of other age groups. Sex had effect only on total erythrocyte counts, mean corpuscular hemoglobin concentration and mean corpuscular hemoglobin in horses of 1-3 years age group (young stock) and on packed cell volume in adult female and male equids. Among biochemical indices, activities of enzymes were observed to be influenced both by age and sex. Creatine kinase, gamma glutamyl transferase, glutamate pyruvate transaminase, glutamic oxaloacetate transaminase and lactate dehydrogenase activities were significantly higher in young and adult equids than animals of other age groups in Kathiawari horses while activity of alkaline phosphatase was significantly higher in yearlings than equids belonging to other age groups in both male and females. However, activity of sorbitol dehydrogenase was unaltered due to both sex and age factor. Albumin, bilirubin direct, bilirubin total, cholesterol, creatinine, protein, triglyceride and uric acid were statistically different in various age and sex groups of horses. Calcium, magnesium and chloride contents were almost same in various age groups of male horses. Significantly higher levels of $T_3$ and $T_4$ were observed in both male and female yearlings as compared to equids of other age groups in both the sexes.

• 대한수의학회지
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• 제45권2호
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• pp.155-161
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• 2005

Proteomics is a useful approach to know protein expression, post-translational modification and protein function. We investigated the protein expression pattern and identity in chickens fed with the tissue culture medium waste after harvest of Korean wild ginseng (TCM-KWG) (Panax ginseng). Two groups (n=60/group) of day old broiler chickens were administered with 0 (control) and 0.8% (treatment) TCM-KWG through drinking water. After 5 weeks, we examined the protein expression pattern of fibularis longus and superficial pectoral muscle by Two-dimensional electrophoresis analysis. Interestingly, TCM-KWG treatment significantly increased five spot's density, and markedly reduced five spot's density in the muscles. We identified 10 proteins (desmin, myosin light chain 1, heat shock 25 kDa protein, collapsin response mediator protein-2A, alpha enolase, vimentin, actin alpha 1, my023 protein, pyruvate kinase and troponin T) by the matrix-assisted laser desorption ionization time of flight (MALDI-TOF).

• International Journal of Oral Biology
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• 제30권3호
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• pp.85-90
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• 2005

Homeostatic pH is very important for various cellular processes, including metabolism, survival, and death. An imbalanced-pH might induce cellular acidosis, which is involved in many abnormal events such as apoptosis and malignancy. One of several factors contributing to the onset of metabolic acidosis is the production of lactate and protons by lactate dehydrogenase (LDH) in anaerobic glycolysis. LDH is an important enzyme that catalyzes the reversible conversion of pyruvate to lactate. This study sought to examine whether decreases in extracellular pH induce apoptosis of CHO cells, and to elucidate the role of mitogen-activated protein kinases (MAPKs) in acidification-induced apoptosis. To test apoptotic signaling by acidification we used CHO dhfr cells that were sensitive to acidification, and CHO/anti-LDH cells that are resistant to acidification-induced apoptosis and have reduced LDH activity by stable LDH antisense mRNA expression. In the present study, cellular lactic acid-induced acidification and the role of MAPKs signaling in acidification-induced apoptosis were investigated. Acidification, which is caused by $HCO{_3}^-$-free conditions, induced apoptosis and MAPKs (ERK, JNK, and p38) activation. However, MAPKs were slightly activated in acidic conditions in the CHO/anti-LDH cells, indicating that lactic acid-induced acidification induces activation of MAPKs. Treatment with a p38 inhibitor, PD169316, increased acidification-induced apoptosis but apoptosis was not affected by inhibitors for ERK (U0126) or JNK (SP600125). Thus, these data support the hypothesis that activation of the p38 MAPK during acidification-induced apoptosis contributes to cell survival.

• 한국수정란이식학회지
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• 제22권4호
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• pp.213-218
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• 2007

This study was performed to investigate the effect of VEGF on in vitro maturation of porcine oocytes. The base medium for IVM, TCM-199 was supplemented with 0.6 mM cysteine, 0.91 mM pyruvate, 10 ng/ml epidermal growth factor, $75{\mu}g/ml$ kenamycin, $1{\mu}g/ml$ insulin and 10% (V/V) porcine follicular fluid (pFF) as a Group A; Group B was consists of Group A plus 5 ng/ml VEGF; Group C was consists of replacement of pFF by 10% PVA and Group D: was consists of Group C plus 5 ng/ml VEGF. 1. The maturation rate was significantly higher (p<0.05) in control and VEGF+pFF group than other two groups ($76.1{\pm}9.6,\;78.9{\pm}6.0\;vs\;60.4{\pm}14.2\;and\;58.3{\pm}14.3$, respectively). 2. Addition of VEGF without pFF showed a negative effect on oocytes maturation and about 58.26% oocytes were reached to M-II stage. 3. In the parthenogenetic development, the cleavage rate was significantly higher (p<0.05) in control and VEGF+pFF group ($73.2{\pm}1.8\;and\;64.6{\pm}1.1$, respectively) than other groups ($47.9{\pm}1.8\;and\;48.3{\pm}1.7$, respectively). 4. The blastocyst formation rate was significantly higher (p<0.05) in VEGF+pFF group ($32.6{\pm}2.4$) compared to control and other groups. 5. There was no significant difference in cell numbers (inner cell mass or trophectoderm) among these groups.

• Archives of Plastic Surgery
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• 제37권4호
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• pp.317-322
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• 2010

Purpose: There is no clear evidence of the original cause of hypertrophic scar, and the effective method of treatment is not yet established. Recently the steps of searching in gene and molecular level are proceeding. we are trying to recognize the difference between keratinocytes of hypertrophic scar and normal skin. Then we do support the comprehension of the scar formation mechanism and scar management. Methods: Total RNAs were extracted from cultured keratinocytes from 4 hypertrophic scars and normal skins. The cDNA chips were prepared. A total of 3063 cDNAs from human cDNA library were arrayed. And the scanning data were analyzed. Results: On microarray, heat shock protein, pyruvate kinase, tumor rejection antigen were more than 2 fold intensity genes. Among them, heat shock 70 kd protein showed the strongest intensity difference. Conclusion: In this study, it can be concluded that heat shock proteins play an important role in the process of wound healing and scar formation. This study provides basic biologic information for scar research. The new way of the prevention and treatment of scar formation would be introduced with further investigations.

• Mass Spectrometry Letters
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• 제9권1호
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• pp.24-29
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• 2018

DC23, a triazolothione resorcinol analogue, is known to inhibit heat shock protein 90 and pyruvate dehydrogenase kinase which are up-regulated in cancer and diabetes, respectively. This study was performed to elucidate the metabolism of DC23 in human liver microsomes (HLMs). HLMs incubated with DC23 in the presence of uridine 5'-diphosphoglucuronic acid (UDPGA) and/or ${\beta}$-nicotinamide adenine dinucleotide phosphate (NADPH) resulted in the formation of four metabolites, M1-M4. M1 was identified as DC23-N-Oxide, on the basis of LC-MS/MS analysis. DC23 was further metabolized to its glucuronide conjugates (M2, M3, and M4). In vitro metabolic stability studies conducted with DC23 in HLMs revealed significant glucuronide conjugation with a $t_{1/2}$ value of 1.3 min. The inhibitory potency of DC23 on five human cytochrome P450s was also investigated in HLMs. In these experiments, DC23 inhibited CYP2C9-mediated tolbutamide hydroxylase activity with an $IC_{50}$ value of $8.7{\mu}M$, which could have implications for drug interactions.