• Molecules and Cells
• /
• v.40 no.6
• /
• pp.393-400
• /
• 2017

Type 2 diabetes mellitus (T2DM) is a chronic metabolic disease characterized by lack of insulin and high glucose levels. T2DM can cause bone loss and fracture, thus leading to diabetic osteoporosis. Promoting osteogenic differentiation of osteoblasts may effectively treat diabetic osteoporosis. We previously reported that Sirtuin 1 (Sirt1), a $NAD^+$-dependent deacetylase, promotes osteogenic differentiation through downregulation of peroxisome proliferator-activated receptor (PPAR) ${\gamma}$. We also found that miR-132 regulates osteogenic differentiation by downregulating Sirt1 in a $PPAR{\beta}/{\delta}$-dependent manner. The ligand-activated transcription factor, $PPAR{\alpha}$, is another isotype of the peroxisome proliferator-activated receptor family that helps maintain bone homeostasis and promot bone formation. Whether the regulatory role of $PPAR{\alpha}$ in osteogenic differentiation is mediated via Sirt1 remains unclear. In the present study, we aimed to determine this role and the underlying mechanism by using high glucose (HG) and free fatty acids (FFA) to mimic T2DM in MC3T3-E1 cells. The results showed that HG-FFA significantly inhibited expression of $PPAR{\alpha}$, Sirt1 and osteogenic differentiation, but these effects were markedly reversed by $PPAR{\alpha}$ overexpression. Moreover, siSirt1 attenuated the positive effects of $PPAR{\alpha}$ on osteogenic differentiation, suggesting that $PPAR{\alpha}$ promotes osteogenic differentiation in a Sirt1-dependent manner. Luciferase activity assay confirmed interactions between $PPAR{\alpha}$ and Sirt1. These findings indicate that $PPAR{\alpha}$ promotes osteogenic differentiation via the Sirt1-dependent signaling pathway.

• Nutrition Research and Practice
• /
• v.13 no.3
• /
• pp.205-213
• /
• 2019

BACKGROUND/OBJECTIVES: Myocardial infarction (MI) is caused by extensive myocardial damage attributed to the occlusion of coronary arteries. Our previous study in a rat model of ischemia/reperfusion (I/R) demonstrated that administration of arabinoxylan (AX), comprising arabinose and xylose, protects against myocardial injury. In this study, we undertook to investigate whether psyllium seed husk (PSH), a safe dietary fiber containing a high level of AX (> 50%), also imparts protection against myocardial injury in the same rat model. MATERIALS/METHODS: Rats were fed diets supplemented with PSH (1, 10, or 100 mg/kg/d) for 3 d. The rats were then subjected to 30 min ischemia through ligation of the left anterior descending coronary artery, followed by 3 h reperfusion through release of the ligation. The hearts were harvested and cut into four slices. To assess infarct size (IS), an index representing heart damage, the slices were stained with 2,3,5-triphenyltetrazolium chloride (TTC). To elucidate underlying mechanisms, Western blotting was performed for the slices. RESULTS: Supplementation with 10 or 100 mg/kg/d of PSH significantly reduces the IS. PSH supplementation (100 mg/kg/d) tends to reduce caspase-3 generation and increase BCL-2/BAX ratio. PSH supplementation also upregulates the expression of nuclear factor erythroid 2-related factor 2 (NRF2), and its target genes including antioxidant enzymes such as glutathione S-transferase mu 2 (GSTM2) and superoxide dismutase 2 (SOD2). PSH supplementation upregulates some sirtuins ($NAD^+$-dependent deacetylases) including SIRT5 (a mitochondrial sirtuin) and SIRT6 and SIRT7 (nuclear sirtuins). Finally, PSH supplementation upregulates the expression of protein kinase A (PKA), and increases phosphorylated cAMP response element-binding protein (CREB) (pCREB), a target protein of PKA. CONCLUSIONS: The results from this study indicate that PSH consumption reduces myocardial I/R injury in rats by inhibiting the apoptotic cascades through modulation of gene expression of several genes located upstream of apoptosis. Therefore, we believe that PSH can be developed as a functional food that would be beneficial in the prevention of MI.

• Journal of Yeungnam Medical Science
• /
• v.9 no.1
• /
• pp.90-101
• /
• 1992

In rat kidney, the changes in concentrations of nucleotides and their derivatives during ischemia induced by renal artery ligation was measured quantitatively with high performance liquid chromatography(HPLC). After the ligation of renal artery for 60minutes, the concentrations of the nucleotides and derivatives were measured. In ischemic tissue, IDP was significantly decreased from $217.4{\pm}12.68{\mu}g$ in control to $80.7{\pm}18.39{\mu}g$ (p<0.01) ; ATP, $307.2{\pm}56.63{\mu}g$ to $47.6{\pm}5.95{\mu}g$ (p<0.01) ; ADP+AMP, $227.1{\pm}7.98{\mu}g$ to $61.4{\pm}3.92{\mu}g$(P<0.01); $NAD^+$, $217.9{\pm}4.49{\mu}g$ to $126.6{\pm}10.44{\mu}g$(P<0.01) ; GTP, $202.5{\pm}23.76{\mu}g$ to $117.7{\pm}14.24{\mu}g$ (P<0.05) ; GMP, $54.5{\pm}9.03{\mu}g$ to $23.7{\pm}0.46{\mu}g$(p<0.05), and inosine, $16.6{\pm}3.45{\mu}g$ to $7.8{\pm}0.87{\mu}g$ (P<0.05). But hypoxanthine and xanthine were significantly increased from $113.0{\pm}15.58{\mu}g$ to $159.7{\pm}12.07{\mu}g$ (P<0.05) and from $87.7{\pm}6.77{\mu}g$ to $173.1{\pm}12.52{\mu}g$ (P<0.01). In ischemic kidney, concentration of ATP was decreased to 39.9% of control at 10 minutes, 19.8% at 30 minutes, and 15.5% at 60 minutes, and ADP+AMP were decreased to 70.3% of control at 10 minutes, 67.3% at 30 minutes, and to 27.0% at 60 minutes, but hypoxanthine and xanthine were increased to 121.5% and 127.1% at 10 minutes, 126.0% and 174.4% at 30 minutes, and 141.4% and 197.3% at 60 minutes Total adenosine nucleotides were decreased to 20.3% of control during 60 minutes of ischemia, but hypoxanthine and xathine were increased to 157.5 % of control. These results suggest that the changes in the concentration of nucleotides and their metabolic derivatives are useful indices of the extents of tissue ischemia in rat kidney.

• Journal of Microbiology and Biotechnology
• /
• v.24 no.12
• /
• pp.1636-1643
• /
• 2014

3-Hydroxybutyryl-CoA dehydrogenase is an enzyme that catalyzes the second step in the biosynthesis of n-butanol from acetyl-CoA, in which acetoacetyl-CoA is reduced to 3-hydroxybutyryl-CoA. To understand the molecular mechanisms of n-butanol biosynthesis, we determined the crystal structure of 3-hydroxybutyryl-CoA dehydrogenase from Clostridium butyricum (CbHBD). The monomer structure of CbHBD exhibits a two-domain topology, with N- and C-terminal domains, and the dimerization of the enzyme was mostly constituted at the C-terminal domain. The mode of cofactor binding to CbHBD was elucidated by determining the crystal structure of the enzyme in complex with $NAD^+$. We also determined the enzyme's structure in complex with its acetoacetyl-CoA substrate, revealing that the adenosine diphosphate moiety was not highly stabilized compared with the remainder of the acetoacetyl-CoA molecule. Using this structural information, we performed a series of site-directed mutagenesis experiments on the enzyme, such as changing residues located near the substrate-binding site, and finally developed a highly efficient CbHBD K50A/K54A/L232Y triple mutant enzyme that exhibited approximately 5-fold higher enzyme activity than did the wild type. The increased enzyme activity of the mutant was confirmed by enzyme kinetic measurements. The highly efficient mutant enzyme should be useful for increasing the production rate of n-butanol.

• Toxicological Research
• /
• v.3 no.2
• /
• pp.97-110
• /
• 1987

The effects of 5 novel hepatotrophic agents, dithiol malonate derivatives (DMDs; DMD1-DMD5), on the liver microsomal lipid peroxidation induced by carbon tetrachloride $(CCl_4)$ and the correlations with the changes of microsomal electron transport system were investigated. All DMDs were found to inhibit the lipid peroxidation induced by $CCl_4$ in mice and rats as well in vitro liver microsomal system. Therefore, each DMD seemed to have direct mode of action on liver microsomes to inhibit the lipid peroxidation. As an ex vivo study, the induced lipid peroxidation by $CCl_4$ and the changes in electron transport system were determined with liver microsomes obtained from rats chronically treated with DMDs for 7 days. The induced lipid peroxide contents in liver microsomal system were lower in DMD1, DMD2 and DMD3 treated group, but higher in DMD4 and DMD5 group when compared to the control group. Cyt. p.450 contents in the microsomes were decreased by the treatment with DMD1, DMD2 and DMD3, but increased significantly by DMD4 with great extent and by DMD5 with less extent. The cyt. p-450 isozymes induced by treatment of DMD4 and DMD5 were identified as 3-methylcholanthrene (MC) type. The NADPH cyt. -C reductase activities of the microsomes treated with DMD1, DMD2, DMD4 and DMD5 were increased in the range of around 20% to 50%, but decreased with DMD3, All DMDs increased dyt. $-b_5$ content and did not alter NAdH-cyt, $-b_5$ reductase activities in the microsomes. In summary, the 5 novel hepatotrophic agents (DMDs) markedly protected against lipid peroxidation induced by $CCl_4$ in vivo and in vitro possibly through the mechanism of direct action on the liver microsomes. The degree of inhibition produced by DMDs on lipid peroxidation induced by $CCl_4$ seemed to coincide rather with cyt. p-450 contents than with other components of liver microsomal electron transport system including NADPH-cyt, -C reductase.

• Biomolecules & Therapeutics
• /
• v.24 no.5
• /
• pp.495-500
• /
• 2016

This study aimed to explore the neuroprotection and mechanism of isoflurane on rats with spinal cord ischemic injury. Total 40 adult male Sprague-Dawley rats were divided into the four groups (n=10). Group A was sham-operation group; group B was ischemia group; group C was isoflurane preconditioning group; group D was isoflurane preconditioning followed by ischemia treatment group. Then the expressions of TWIK-related $K^+$ channel 1 (TREK1) in the four groups were detected by immunofluorescent assay, real time-polymerase chain reactions (RT-PCR) and western blot. The primary neurons of rats were isolated and cultured under normal and hypoxic conditions. Besides, the neurons under two conditions were transfected with green fluorescent protein (GFP)-TREK1 and lentivirual to overexpress and silence TREK1. Additionally, the neurons were treated with isoflurane or not. Then caspase-3 activity and cell cycle of neurons under normal and hypoxic conditions were detected. Furthermore, nicotinamide adenine dinucleotide hydrate (NADH) was detected using NAD+/NADH quantification colorimetric kit. Results showed that the mRNA and protein expressions of TREK1 increased significantly in group C and D. In neurons, when TREK1 silenced, isoflurane treatment improved the caspase-3 activity. In hypoxic condition, the caspase-3 activity and sub-G1 cell percentage significantly increased, however, when TREK1 overexpressed the caspase-3 activity and sub-G1 cell percentage decreased significantly. Furthermore, both isoflurane treatment and overexpression of TREK1 significantly decreased NADH. In conclusion, isoflurane-induced neuroprotection in spinal cord ischemic injury may be associated with the up-regulation of TREK1.

• Journal of The Korean Society of Grassland and Forage Science
• /
• v.10 no.2
• /
• pp.102-109
• /
• 1990

This experiment was carried out to study the effect of pure mineral nitrogen fertilizing on dry matter yield of grassland and the advisable mineral nitrogen amounts in long duration under practical conditions at the "Federal Institute for Agriculture in the Alps" in Austria. The application rates were 0, 30, 60,90 and 120 kg N/ha/cut, the cutting regimes 3-, 4-, 5- and 6-cuts/year. In order to explain the nitrogen-profitability were determined that 1 kg pure mineral nitrogen have to produce 8 - 16 kg DM/kg N in dependence on cutting regimes and requiring of nitrogen efficiencies. The results were as follows: 1. With only PK-fertilizaing average dry matter yields from 4.0 to 7.6 tons per ha and year were obtained. 2. Within all applied cutting regimes 60 kg min. Nlhalgrowth have proved to be the most efficient application rate witn 13 - 24 kg DM/kg N in dependent of cutting regimes. Comapred with only PK-treatment the DM yields increased by 3.9 - 4.7 t/ ha nad year. 3. By the sigmaformed process of Input-Output curve the highest marginal yield (the "most efficient" Ndressing rate) per ha and year was calculated: 152 kg N at 3-cut regimes, 204 kg N at 4-cut regimes, 220 kg N at 5-cut regimes and 240 kg N/ha/year at 6-cut regimes. 4. With required efficiencies of 16 and 12 kg DM/kg N 240 - 300 kg N per ha and year respectively would have to be applied at 3-cut regimes; with required efficiencies of 12 and 10 kg DM/kg N at 4-cut regimes the appropriate figures ranged from 320 to 420 kg N/ha and year, at 5- and 6-cut regimes and efficiencies of 10 and 8 kg DM/kg N results of 360 - 460 kg N and 380 - 500 kg N respectively were obtained. 5. At the relatively dry location Piber the highest dressing rates were needed in order to obtain the efficiencies from 8 to 16 kg DM/kg N, about 30 - 60 kg N/ha/year more than at the relatively moist location Admont.ist location Admont.

• The Journal of Korean Obstetrics and Gynecology
• /
• v.35 no.3
• /
• pp.1-23
• /
• 2022

Objectives: The purpose of this study is to evaluate anti-breast cancer and anti-inflammatory effects of Chungganhaewool-tang and Shipyeukmiyeugi-eum. Methods: MDA-MB-231 cells were used to measure cytotoxicity, Reactive oxygen species (ROS) production, protein expression amounts of Bcl-2-associated X protein (Bax), B-cell lymphoma 2 (Bcl-2), B-cell lymphoma-extra large (Bcl-xl), Cytochrome C Caspase-3, Caspase-7, Caspase-9, Poly ADP-ribose polymerase (PARP), Nuclear factor erythroid-2-related factor 2 (Nrf2), Heme oxygenase-1 (HO-1) and NAD (P) H Quinone Oxidoreductase 1 (NQO1) to evaluate the anti-breast cancer effects of Chungganhaewool-tang (CHT) and Shipyeukmiyeugi-eum (SYE), and THP-1 cells, differentiated into macrophage and induced inflammation with Lipopolysaccharide (LPS), were used to measure production amounts of ROS, Nitric oxide (NO), and protein expression amounts of Inducible nitric oxide synthase (iNOS), Cyclooxygenase (COX-2), Interleukin-1 beta (IL-1β), Interleukin-6 (IL-6) and Tumor necrosis factor-alpha (TNF-α) to evaluate the anti-inflammatory effects of CHT and SYE. Results: CHT and SYE reduced MDA-MB-231 cell counts, increased protein expression of Bax and Cytochrome C, and decreased protein expression of Bcl-2, Bcl-xl. The protein expression amounts of Caspase-3, 7, and 9 decreased, but amounts of the active form, cleaved Caspase-3, 7, and 9, increased. In addition, PARP protein expression decreased, the amount of PARP protein in the cleaved form increased, and the amount of protein expressions of Nrf2 and HO-1 decreased, but NQO1 showed no significant difference. In THP-1 cells CHT and SYE reduced ROS and NO, and reduced protein expressions of iNOS, COX-2, IL-1, and TNF-α, but only SYE groups reduced IL-6. Conclusions: This study suggests that CHT and SYE have potential to be used as treatments for breast cancer.

• Journal of Microbiology and Biotechnology
• /
• v.15 no.1
• /
• pp.80-85
• /
• 2005

Electrochemical control of the metabolic flux of W. kimchii sk10 on glucose and pyruvate was studied. The growing cell of W. kimchii sk10 produced 87.4 mM lactate, 69.3 mM ethanol, and 4.9mM lactate from 83.1mM glucose under oxidation condition of the anode compartment, but 98.9 mM lactate, 84.3mM ethanol, and 0.2 mM acetate were produced from 90.8 mM glucose under reduction condition of the cathode compartment for 24 h, respectively. The resting cell of W. kimchii sk10 produced 15.9 mM lactate and 15.2 mM acetate from 32.1 mM pyruvate under oxidation condition of the anode compartment, and 71.3 mM lactate and 3.8 mM acetate from 79.8mM pyruvate under reduction condition of the cathode compartment. The redox balance (NADH/$NAD^+$) of metabolites electrochemically produced from pyruvate was 1.05 and 18.76 under oxidation and reduction conditions, respectively. On the basis of these results, we suggest that the neutral red (NR) immobilized in bacterial membrane can function as an electron channel for the electron transfer between electrode and cytoplasm without dissipation of membrane potential, and that the bacterial fermentation of W. kimchii sk10 can be shifted to oxidized or reduced pathways by the electrochemical oxidation or reduction, respectively.

• Asian-Australasian Journal of Animal Sciences
• /
• v.21 no.11
• /
• pp.1673-1679
• /
• 2008

In order to study the biological function of gapdh gene in yak, and prove whether the gapdh gene was a useful intra-reference gene that can be given an important role in molecular biology research of yak, the cDNA sequence encoding glyceraldehyde-3-phosphate dehydrogenase from yak was cloned by the RT-PCR method using gene specific PCR primers. The sequence results indicated that the cloned cDNA fragment (1,008 bp) contained a 1,002 bp open reading frame, encoding 333 amino acids (AAs) with a molecular mass of 35.753 kDa. The deduced amino acids sequence showed a high level of sequence identity to Bos Taurus (99.70%), Xenopus laevis (94.29%), Homo sapiens (97.01%), Mus musculus (97.90%) and Sus scrofa (98.20%). The expression of yak's gapdh gene in heart, spleen, kidney and brain tissues was also detected; the results showed that the gapdh gene was expressed in all these tissues. Further analysis of yak GAPDH amino acid sequence implied that it contained a complete glyceraldehyde-3-phosphate dehydrogenase active site (ASCTTNCL) which ranged from 148 to 155 amino acid residues. It also contained two conserved domains, a NAD binding domain in its N-terminal and a complete catalytic domain of sugar transport in its C-terminal. The phylogenetic analysis showed that yak and Bos taurus were the closest species. The prediction of secondary structures indicated that GAPDH of yak had a similar secondary structure to other isolated GAPDH. The results of this study suggested that the gapdh gene of yak was similar to other species and could be used as the intra-reference to analyze the expression of other genes in yak.