Reactive oxygen species (ROS) and nitrogen species (RNS) are both important signaling molecules involved in pain transmission in the dorsal horn of the spinal cord. Xanthine oxidase (XO) is a well-known enzyme for the generation of superoxide anions ($O_2^{\bullet-}$), while S-nitroso-N-acetyl-DL-penicillamine (SNAP) is a representative nitric oxide (NO) donor. In this study, we used patch clamp recording in spinal slices of rats to investigate the effects of $O_2^{\bullet-}$ and NO on the excitability of substantia gelatinosa (SG) neurons. We also used confocal scanning laser microscopy to measure XO- and SNAP-induced ROS and RNS production in live slices. We observed that the ROS level increased during the perfusion of xanthine and xanthine oxidase (X/XO) compound and SNAP after the loading of 2',7'-dichlorofluorescin diacetate ($H_2DCF-DA$), which is an indicator of intracellular ROS and RNS. Application of ROS donors such as X/XO, ${\beta}-nicotinamide$ adenine dinucleotide phosphate (NADPH), and 3-morpholinosydnomimine (SIN-1) induced a membrane depolarization and inward currents. SNAP, an RNS donor, also induced membrane depolarization and inward currents. X/XO-induced inward currents were significantly decreased by pretreatment with phenyl N-tert-butylnitrone (PBN; nonspecific ROS and RNS scavenger) and manganese(III) tetrakis(4-benzoic acid) porphyrin (MnTBAP; superoxide dismutase mimetics). Nitro-L-arginine methyl ester (NAME; NO scavenger) also slightly decreased X/XO-induced inward currents, suggesting that X/XO-induced responses can be involved in the generation of peroxynitrite ($ONOO^-$). Our data suggest that elevated ROS, especially $O_2^{\bullet-}$, NO and $ONOO^-$, in the spinal cord can increase the excitability of the SG neurons related to pain transmission.
This study examined the effects of black ginseng (9 times-steamed ginseng) on hypoglycemic action in streptozotocininduced diabetic rats as well as changes in ginsenoside composition by the steaming process. As the number of steaming cycles increased, the amounts of crude saponin and most ginsenoside contents decreased, while the amount of ginsenoside- Rg3 and the ratio of PD/PT (=[$Rb_1+Rb_2+Rc+Rd+Rg_3]/[Re+Rb_1+Rh_1]$) increased. This ginsenoside composition is a unique characteristic compared to other types of ginseng products. In order to investigate the hypoglycemic effect of the black ginseng extract, in vivo studies were performed in rats with streptozotocin-induced diabetes. The studies showed that the administration of the black ginseng extract decreased high blood glucose levels (more than 300 mg/dL) to a normal level (102 mg/dL). These results suggest that this black ginseng extract has a significant hypoglycemic effect and can be used as an anti-diabetic substance for dietary supplements or new drugs.
N-acetylglucosamine kinase (GlcNAc kinase or NAGK; EC 2.7.1.59) is highly expressed and plays a critical role in the development of dendrites in brain neurons. In this study, the authors conducted structure-function analysis to verify the previously proposed 3D model structure of GlcNAc/ATP-bound NAGK. Three point NAGK mutants with different substrate binding capacities and reaction velocities were produced. Wild-type (WT) NAGK showed strong substrate preference for GlcNAc. Conversion of Cys143, which does not make direct hydrogen bonds with GlcNAc, to Ser (i.e., C143S) had the least affect on the enzymatic activity of NAGK. Conversion of Asn36, which plays a role in domain closure by making a hydrogen bond with GlcNAc, to Ala (i.e., N36A) mildly reduced NAGK enzyme activity. Conversion of Asp107, which makes hydrogen bonds with GlcNAc and would act as a proton acceptor during nucleophilic attack on the ${\gamma}$-phosphate of ATP, to Ala (i.e., D107A), caused a total loss in enzyme activity. The overexpression of EGFP-tagged WT or any of the mutant NAGKs in rat hippocampal neurons (DIV 5-9) increased dendritic architectural complexity. Finally, the overexpression of the small, but not of the large, domain of NAGK resulted in dendrite degeneration. Our data show the effect of structure on the functional aspects of NAGK, and in particular, that the small domain of NAGK, and not its NAGK kinase activity, plays a critical role in the upregulation of dendritogenesis.
This study determined the effects of fucoxanthin on gene expressions related to lipid metabolism in rats with a high-fat diet. Rats were fed with normal fat diet (NF, 7% fat) group, high fat diet group (HF, 20% fat), and high fat with 0.2% fucoxanthin diet group (HF+Fxn) for 4 weeks. Body weight changes and lipid profiles in plasma, liver, and feces were determined. The mRNA expressions of transcriptional factors such as sterol regulatory element binding protein (SREBP)-1c, Carnitine palmitoyltransferase-1 (CPT1), Cholesterol $7{\alpha}$-hydroxylase1 (CYP7A1) as well as mRNA expression of several lipogenic enzymes were determined. Fucoxanthin supplements significantly increased plasma high density lipoprotein (HDL) concentration (P < 0.05). The hepatic total lipids, total cholesterols, and triglycerides were significantly decreased while the fecal excretions of total lipids, cholesterol, and triglycerides were significantly increased in HF+Fxn group (P < 0.05). The mRNA expression of hepatic Acetyl-CoA carboxylase (ACC), Fatty acid synthase (FAS), and Glucose-6-phosphate dehydrogenase (G6PDH) as well as SREBP-1C were significantly lower in HF+Fxn group compared to the HF group (P < 0.05). The hepatic mRNA expression of Hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) and Acyl-CoA cholesterol acyltransferase (ACAT) were significantly low while lecithin-cholesterol acyltransferase (LCAT) was significantly high in the HF+Fxn group (P < 0.05). There was significant increase in mRNA expression of CPT1 and CYP7A1 in the HF+Fxn group, compared to the HF group (P < 0.05). In conclusion, consumption of fucoxanthin is thought to be effective in improving lipid and cholesterol metabolism in rats with a high fat diet.
Nicotinamide adenine dinucleotide phosphate oxidases (NOXs) are the major enzymatic source of reactive oxygen species (ROS). NOX2 and NOX4 are expressed in the heart but its role in hypoxia-induced atrial natriuretic peptide (ANP) secretion is unclear. This study investigated the effect of NOX on ANP secretion induced by hypoxia in isolated beating rat atria. The results showed that hypoxia significantly upregulated NOX4 but not NOX2 expression, which was completely abolished by endothelin-1 (ET-1) type A and B receptor antagonists BQ123 (0.3 μM) and BQ788 (0.3 μM). ET-1-upregulated NOX4 expression was also blocked by antagonists of secreted phospholipase A2 (sPLA2; varespladib, 5.0 μM) and cytosolic PLA2 (cPLA2; CAY10650, 120.0 nM), and ET-1-induced cPLA2 expression was inhibited by varespladib under normoxia. Moreover, hypoxia-increased ANP secretion was evidently attenuated by the NOX4 antagonist GLX351322 (35.0 μM) and inhibitor of ROS N-Acetyl-D-cysteine (NAC, 15.0 mM), and hypoxia-increased production of ROS was blocked by GLX351322. In addition, hypoxia markedly upregulated Src expression, which was blocked by ET receptors, NOX4, and ROS antagonists. ET-1-increased Src expression was also inhibited by NAC under normoxia. Furthermore, hypoxia-activated extracellular signal-regulated kinase 1/2 (ERK1/2) and protein kinase B (Akt) were completely abolished by Src inhibitor 1 (1.0 μM), and hypoxia-increased GATA4 was inhibited by the ERK1/2 and Akt antagonists PD98059 (10.0 μM) and LY294002 (10.0 μM), respectively. However, hypoxia-induced ANP secretion was substantially inhibited by Src inhibitor. These results indicate that NOX4/Src modulated by ET-1 regulates ANP secretion by activating ERK1/2 and Akt/GATA4 signaling in isolated beating rat hypoxic atria.
Objective: This study was conducted to investigate the potential effects of prolonged photoperiod on the serum lipids, carcass traits, and meat quality of Jinjiang cattle during winter. Methods: Thirty-four Jinjiang bulls aged between 14 and 16 months were randomly assigned to two groups that were alternatively subjected to either natural daylight +4 h supplemental light (long photoperiod, LP) or natural daylight (natural photoperiod, NP) for 96 days. The potential effects on the levels of serum lipids, carcass traits, meat quality, and genes regulating lipid metabolism in the intramuscular fat (IMF) of the cattle were evaluated. Results: Jinjiang cattle kept under LP showed significant increase in both dry matter intake and backfat thickness. the serum glucose and the plasma leptin levels were significantly reduced, while that of melatonin and insulin were observed to be increased. The crude fat contents of biceps femoris muscle and longissimus dorsi muscle were higher in LP than in NP group. In longissimus dorsi muscle, the proportions of C17:0 and C18:0 were significantly higher but that of the C16:1 was found to be significantly lower in LP group. The relative mRNA expressions in IMF of longissimus dorsi muscle, the lipid synthesis genes (proliferator-activated receptor gamma, fatty acid-binding protein) and the fatty acid synthesis genes (acetyl-coa carboxylase, fatty acid synthetase, 1-acylglycerol-3-phosphate acyltransferase) were significantly up-regulated in LP group (p<0.05); whereas the hormone-sensitive lipase and stearoyl-CoA desaturase 1 were significantly down-regulated in LP than in NP group. Conclusion: Prolonged photoperiod significantly altered the growth performance, hormonal levels, gene expression and fat deposition in Jinjiang cattle. It suggested that the LP improved the fat deposition by regulating the levels of different hormones and genes related to lipid metabolism, thereby improving the fattening of Jinjiang cattle during winter.
Hangover after drinking is different from person to person symptoms and degree, but usually thirst, fatigue, headache, general boredom, gastrointestinal disorder, vomiting, diarrhea, deficiency of vitamin appears. This hanging phenomenon is caused by the action of precursors such as ethyl acetate and acetaldehyde, which are the by products of fermentation contained in alcohol and alcohol accumulated in hepatocytes and body. In order to solve the hangover phenomenon, the same Origin as polysaccharide Polydeoxyribonucleotide, which is a nucleic acid-sugar-phosphate complex, which is a semen or testicular extract in salmon extract, and a water soluble salmon extract powder having the same structure and lower price than Polydeoxyribonucleotide And D-Glucuronic acid and N-Acetyl glucosamine. It has excellent biocompatibility, viscoelasticity and moisturizing power. It has effect on reduction of body water loss and skin moisture content in hangover phenomenon. It is antioxidant and skin moisturizing effect Hyaluronic acid was irradiated with gamma rays, and the composition was prepared by using the salmon extract powder and the main raw material. The ethanol degradation, the acetaldehyde reduction amount, the blood acetaldehyde concentration and the acetic acid concentration were measured to evaluate the alcoholysis effect, Skin moisture evaporation rate To examine the evaluation unit water content of the skin was improved determine whether the antioxidant and provide skin moisturizing effect. The addition of ethanol extracts of salmon extracts showed a decrease of 5 to 7 times compared with no addition, and a decrease of 3 to 5 times of acetaldehyde. In addition, the change of acetaldehyde concentration and acetic acid concentration in blood showed a rapid decrease compared to the no - added control group. In addition, when the raw material of hyaluronic acid was used, skin moisture content was high and skin moisture evaporation amount was decreased. Therefore, hyaluronic acid, which is a polysaccharide polymer, has excellent viscoelasticity and moisturizing ability, It is considered to provide antioxidant and skin moisturizing effect. Therefore, it can be said that the composition containing salmon extract powder and hyaluronic acid as a main ingredient is effective for the hangover phenomenon which occurs after drinking.
Present experiment was performed to establish the optimal reaction conditions for measurement of urinary gamma-glutamyltranspeptidase(${\gamma}$-GTP), N-acetyl-${\beta}$-D-glucosaminidase (AGS) and alanine aminopeptidase(AAP) activities in bovine and to investigate in vitro stability of the enzymes, within-run imprecision of the methods, and normal ranges. 1. The optimal wavelength for measurement of ${\gamma}$-GTP activity was 545nm. 2. The optimal pH of Tris-HCI buffer containing glycylglycine for measurement of urinary ${\gamma}$-GTP activity was 7.6~7.8(37$^{\circ}C$). 3. Coefficient of variance for within-run imprecision of urinary ${\gamma}$-GTP activity ranged from 4.8 to 7.2% and there was no significant difference among replications, 4. The optimal wavelength for measurement of urinary AGS activity was 405nm. 5. The optimal pH of citrate buffer for measurement urinary of AGS activity was 4.0(37$^{\circ}C$). 6. Coefficient of variance for within-run imprecision of urinary AGS activity ranged from 3.9 to 6.1% and there was no significant difference among replications. 7. The optimal wavelength for measurement of urinary AAP activity was 400nm. 8. The optimal pH of phosphate buffer for measurement of urinary AAP was 7.8. 9. Coefficient of variance for within-run imprecision of urinary AAP activity ranged from 2.5 to 4.8% and there was no significant difference among replications. 10. ${\gamma}$-GTP and AGS activities were increased significantly by gel-filtration. 11. Turbidity interfered with measurement of urinary AAP activity in bovine unless the specimen was gel-filterated. 12. Preservation of the specimen at 5$^{\circ}C$ or -20$^{\circ}C$ did not affect the AGS activity at least for 7 days after collection. 13. Preservation of the specimen at 5$^{\circ}C$ or 20$^{\circ}C$ did not affect the ${\gamma}$-GTP and AAP activities statistically, but some individual specimens revealed fluctuation during preservation. 14. ${\gamma}$-GTP, AGS and AAP activities revealed fluctuation by the tine of the day when the specimen was collected. 15. The normal ranges of urinary ${\gamma}$ -GTP, AGS and AAP activities were 6.60${\pm}$3.26(2.36-14.50), 1.31 ${\pm}$ 0.81(0.33-3.78), and 1.73 ${\pm}$ 0.55(0.77-3.03)U/l. respectively.
In this study, we investigated the anti-obese activity of HPJ extract in C57BL/6J mice. The C57BL/6J mice were randomly divided into five groups: normal control group (Con), high fat diet control group (HFD), treatment groups with HPJ at 125 mg/kg (HPJ125), 250 mg/kg (HPJ250), or 500 mg/kg (HPJ500). To induce an obesity, mice were fed by a high fat diet for 6 weeks, and mice were administered with HPJ extract once a day for 8 weeks. At the end of treatment, we examined the effect of HPJ extract on body weight, plasma lipid, and lipogenic enzymes. HPJ extract was found to lower whole body and epididymal adipose tissue weights and lowered plasma levels of glucose, insulin, triglyceride (TG), total cholesterol (TC), non-esterified fatty acid (NEFA) and leptin, compared to those in HFD group. Histological analyses of the liver and fat tissues of mice treated with HPJ extract revealed significantly decreased number of lipid droplets and decreased size of adipocytes compared to the HFD group. In addition, HPJ extract preserved the morphological integrity of pancreatic islets. To elucidate an action mechanism of HPJ extract, Western blot and RT-PCR were performed using epididymal adipose tissues. HPJ extract up-regulated the levels of phosphorylated adenosine monophosphate-activated protein kinase (AMPK) and its substrate, acetyl-CoA carboxylasse (ACC). HPJ extract also attenuated lipogenic gene expressions of sterol regulatory element-binding protein $1{\alpha}$ (SREBP$1{\alpha}$), fatty acid synthase (FAS), sterol-CoA desaturase 1 (SCD1) and glycerol-3-phosphate acyltransferase (GPAT) in dose-dependent manners. In contrast, expressions of lipolytic genes such as peroxisome proliferator-activated receptor-$\alpha$ (PPAR-${\alpha}$) and CD36, and fatty acid $\beta$-oxidation gene, carnitine palmitoyltransferase-1 (CPT-1) were increased. These results suggest that HPJ extract ameliorates obesity through inhibiting synthesis of lipogenic enzymes as well as stimulating fatty acid oxidation resulting from activation of AMPK, and HPJ extract could be developed as a potential therapeutic agent for obese patients.
The objective of this experiment was to determine the effects of n-3 polyunsaturated fatty acids (n-3 PUFAs)-enriched diet supplemented with different levels of $\alpha$-tocopherol on the activities of hepatic lipogenic-related enzymes and the contents of liver and plasma lipid fractions in laying Tsaiya ducks. A total of 180 30-wk-old laying Tsaiya ducks, at the beginning of peak production, were allotted into 6 treatments with 3 replicates each. Ducks were fed one of the 6 experimental diets, containing 4% tallow (control), and 4% fish oil supplemented with graded levels of $\alpha$-tocopheryl acetate ($\alpha$-tocopherol) at 0, 100, 200, 300 and 400 mg/kg, respectively, for 6 wks. Feed and water were supplied ad libitum throughout the experimental period. The results indicated that the n-3 PUFAsenriched diet supplemented with different levels of $\alpha$-tocopherol did not affect (p>0.05) egg weight, feed intake, body weight change or liver and abdominal fat weights. Egg production, egg mass and feed efficiency significantly (p<0.05) improved as dietary $\alpha$-tocopherol levels increased. The activities of hepatic lipogenic-related enzymes including acetyl-CoA carboxylase (EC 6. 2. 1. 3; ACC), glucose-6-phosphate dehydrogenase (EC 1. 1. 1. 49; G-6-PDH), ATP-citrate cleavage enzyme (EC 4. 1. 3. 8; CCE), NADP-malate dehydrogenase (EC 1.1.1.40; NADP-MDH) and fatty acid synthetase (FAS) were higher (p<0.05) in birds fed with the tallow diet than in those fed with fish oil diets and increased with increasing dietary $\alpha$-tocopherol levels. None of the dietary treatments significantly affected the contents of triglyceride and total cholesterol in the liver, or total cholesterol, phospholipid and total lipid in the plasma. However, the contents of phospholipid and total lipid in the liver, and triglyceride in the plasma increased as dietary $\alpha$-tocopherol levels increased. Increasing dietary $\alpha$-tocopherol levels decreased the non-esterified fatty acid (NEFA) content in the plasma and trended to decrease the cholesterol contents in the egg yolk. The lipid metabolism of laying Tsaiya ducks was influenced not only by the dietary fat but also by the supplementation levels of $\alpha$-tocopherol.
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