Alzheimers disease (AD) is the most prevalent form of neurodegenerations associated with aging in the human population. This disease is characterized by the extracellular deposition of beta-amyloid (A ${\beta}$) peptide in cerebral plaques. The A ${\beta}$ peptide is derived from the ${\beta}$-amyloid precursor protein ( ${\beta}$APP). Photolytic processing of ${\beta}$APP by ${\beta}$-secretase(beta-site APP-cleaving enzyme, BASE) and ${\gamma}$-secretase generates the A ${\beta}$ peptide. Several lines of evidence support that A ${\beta}$-induced neuronal cell death is major mechanisms of development of AD. Accordingly, the ${\beta}$-and ${\gamma}$-secretase have been implicated to be excellent targets for the treatment of AD. We previously found that sesaminol glucosides have improving effect on memory functions through anti-oxidative mechanism. In this study, to elucidate possible other mechanism (inhibition of ${\beta}$-and ${\gamma}$-secretase) of sesaminol glucosides, we examined the improving effect of sesaminol glucosides in the scopolamine (1 mg/kg/mouse)-induced memory dysfunction using water maze test in the mice. Sesaminol glucosides (3.75, 7.5 mg/kg/6ml/day p.o., for 3 weeks) reversed the latency time, distance and velocity by scopolamine in dose dependent manner. Next, ${\beta}$-and ${\gamma}$-secretase activities were determined in different regions of brain. Sesaminol glucosides dose-dependently attenuated scopolamine-induced ${\beta}$-secretase activities in cortex and hippocampous and ${\gamma}$-secretase in cortex. This study therefore suggests that sesaminol glucosides may be a useful agent for prevention of the development or progression of AD, and its inhibitory effect on secretase may play a role in the improving action of sesaminol glucosides on memory function.
This study has investigated the effect of a potent bioflavonoid, troxerutin, on diabetes-induced changes in pro-inflammatory mediators and expression of microRNA-146a and nuclear factor-kappa-B (NF-κB) signaling pathway in aortic tissue of type-I diabetic rats. Male Wistar rats were randomly divided into four groups (n = 6/each): healthy, healthy-troxerutin, diabetic, and diabetic-troxerutin. Diabetes was induced by streptozotocin injection (60 mg/kg; intraperitoneally) and lasted 10 weeks. Troxerutin (150 mg/kg/day) was administered orally for last month of experiment. Inflammatory cytokines IL-1β, IL-6, and TNF-α, as well as intercellular adhesion molecule-1 (ICAM-1), vascular cell adhesion molecule (VCAM), cyclooxygenase-II (COX-II), and inducible-nitric oxide synthase (iNOS) were measured on aortic samples by enzyme-linked immunosorbent assay. Gene expressions for transcription factor NF-κB, interleukin-1 receptor-associated kinase-1 (IRAK-1), TNF receptor-associated factor-6 (TRAF-6), and microRNA-146a were determined using real-time polymerase chain reaction. Ten-week diabetes significantly increased mRNA levels of IRAK-1, TRAF-6, NF-κB, and protein levels of cytokines IL-1β, IL-6, TNF-α, adhesion molecules ICAM-1, VCAM, and iNOS, COX-II, and decreased expression of microRNA-146a as compared with healthy rats (p < 0.05 to p < 0.01). However, one month treatment of diabetic rats with troxerutin restored glucose and insulin levels, significantly decreased expression of inflammatory genes and pro-inflammatory mediators and increased microRNA level in comparison to diabetic group (p < 0.05 to p < 0.01). In healthy rats, troxerutin had significant reducing effect only on NF-κB, TNF-α and COX-II levels (p < 0.05). Beside slight improvement of hyperglycemia, troxerutin prevented the activation of NF-κB-dependent inflammatory signaling in the aorta of diabetic rats, and this response may be regulated by microRNA-146a.
Jo, Hee Geun;Kim, Min Ji;Moon, Bo Yeong;Cheong, Sun Hee
Korean Journal of Exercise Nutrition
/
v.23
no.4
/
pp.6-13
/
2019
[Purpose] In this study, we examined the in vitro antioxidant activities and laxative effects of taurine-xylose (T-X), a synthetic taurine-carbohydrate derivative, in a rat model of constipation induced by loperamide. [Methods] The animals were divided into four treatment groups: normal untreated rats (NOR group), loperamide-treated control rats (CON group), loperamide and taurine-xylose (15 mg/kg)-treated rats (T-X group), and loperamide and commercial Dulcolax S (5.5 mg/kg)-treated rats (DS group). [Results] In the present study, T-X exhibited potent reducing power and free radical scavenging activities for DPPH (1,1-diphenyl-2-picrylhydrazyl) and ABTS+ (2,2'-azino-bis (3-ethylbenzothiazoline 6-sulfonic acid ammonium salt) radicals. The laxative effects of T-X were dependent on food, body weight, fecal properties, gastrointestinal transit (GIT) ratio, and serum metabolic parameters. In the T-X group, the number, wet weight, and water content of fecal pellets were noticeably increased compared to those in the loperamide-induced group. T-X treatment significantly increased the activities of hepatic antioxidant enzymes, including those of glutathione peroxidase (GSH-Px), superoxide dismutase (SOD), and catalase (CAT), relative to those in loperamide-induced constipated rats. Furthermore, the GIT ratio and loperamide-induced metabolic parameters in serum, including gastrin (GAS), motilin (MTL), and somatostatin (SS) levels, were significantly improved by T-X treatment. [Conclusion] These results suggest that taurine-xylose exerts antioxidant activities and laxative effects on loperamide-induced constipation by promoting gastrointestinal motility.
This study investigated the antioxidant and antidiabetic activities of Stachys sieboldii Miq. extracts by solvents (water, ethanol, butanol, chloroform, and hexane). The contents of total polyphenols (7.18-37.25 mg/g) and flavonoids (0.21-5.21 mg/g) in extracts from Stachys sieboldii Miq. showed a significant difference dependent on the extraction solvents, butanol > ethanol > water > chloroform > hexane. Antioxidant activities by DPPH and ABTS radical scavenging were increased in a dose-dependent manner. These activity trends associated with the extraction solvent were different at each concentration, but resembled phenolic compound contents trend, generally. FRAP value increased in a dose-dependent manner, but there was a difference in radical scavenging activities when comparing between extraction solvents by butanol > ethanol > hexane > chloroform > water on all concentrations. The trend of ${\alpha}$-amylase inhibition of extracts from $1,000{\mu}g/mL$ to $2,000{\mu}g/mL$ was not affected as enzyme activity is promoted and not inhibited. The inhibition of ${\alpha}$-glucosidase was increased in a dose-dependent manner without water extracts, the activity on hexane extracts was higher than others per the extraction solvent. ${\alpha}$-Glucosidase inhibition of hexane extracts showed 57.76% at $250{\mu}g/mL$, which is 2.8 times higher than the second highest chloroform extract (20.65%). From these results, we presume that the active ingredients of Stachys sieboldii Miq. is different according to the extraction solvent and also the activity is different by these major functional groups.
Nitric oxide(NO) has recently been shown to play an important role on central nervous system(CNS) function in mammals. It is synthesized from L-arginine by the enzyme NO synthase. In this study, we examined this enzyme's existence in CNS of rainbow trout(Oncorhynchus mykiss) and described the forms of microglia. astrocytes. and oligodendrocytes. Two forms of microglia are distributed in CNS. one resembling their mammalian counterpart(large microglia : LM). and the other comprising small microglia(SM) with very little cytoplasm. CNS contained astrocytes of a distinct type which form reticular network, but lack connections to capillaries. The oligodendrocyte was generally a much denser cell than the astrocyte. We have detected NOS($1.04{\pm}0.12\;pg/min/mg$) from rainbow trout CNS. It could be inhibited reversibly or irreversibly by $N^{G}MMA$ and EGTA. These result suggest that the formation of NO from L-arginine in CNS is calcium-dependent and a pathway of early evolutionary orgin.
Kim, So-Yun;Lee, Hyun-Jung;Park, Jae-Hee;Kim, Rae-Young;Cheong, Hyeonsook;Park, Eunju
Journal of the Korean Society of Food Science and Nutrition
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v.42
no.3
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pp.325-334
/
2013
The objective of this study was to compare the content and metabolic activities between fresh pine needle juice (PNJ) and fermented pine needle juice (FPNJ). A variety of factors were measured, including total phenolic content (TPC), antioxidant activity [DPPH radical scavenging activity (RSA), total radical-trapping antioxidant potential (TRAP), oxygen radical absorbance capacity (ORAC), cellular antioxidant capacity (CAC)], anti-genotoxic activity, ${\alpha}$-glucosidase inhibitory activity, and angiotensin converting enzyme (ACE) inhibitory activity. The TPC was $17.3{\pm}0.2$ and $4.6{\pm}0.0$ mg GAE/g in PNJ and FPNJ, respectively. The DPPH RSA, TRAP, and ORAC values increased in a dose-dependent manner for both PNJ and FPNJ, with significantly higher activities in PNJ than FPNJ. The CAC against AAPH-induced oxidative stress in HepG2 cells was protected by both PNJ and FPNJ. Pretreatment with PNJ and FPNJ in human leukocytes produced significant reductions in $H_2O_2$-induced DNA damage at a concentration of $50{\mu}g/mL$. ${\alpha}$-Glucosidase inhibitory activity was significantly higher in FPNJ than PNJ. The ACE inhibitory activity was about 87.1% and 60.0% in 1:1 diluted PNJ and FPNJ, respectively. This study suggests that the fermentation of PNJ could enhance the regulation of blood glucose metabolism and both PNJ and FPNJ might be a new potential source of natural antioxidant, anti-diabetic, and anti-hypertensive agents applicable to food.
We investigated the effects of immobilization chemistry on the yield of immobilization and the bioactivity of the immobilized enzymes. Trypsin as a model protein and macroporous polymer beads(Toyopearl AF 650M, Tosho Co., Japan) was used as a model matrix. Four methods were used to immobilize trypsin; covalent conjugation by reductive amination(at pH 10.0 and pH 4.0) and affinity interaction via streptavidin-biotin, and double-affinity interaction via biotin-streptavidin-biotin system. The covalent conjugation immobilized $3{\sim}4$ mg/ml-gel, ca. 3-fold higher than the affinity method. However, the specific activity of the covalently(pH 10.0) and affinity-immobilized trypsin(via streptavidin-biotin) are ca. 37% and 50%, respectively, of that of the soluble enzyme(on the low-molecular-weight BAPNA substrate). When the molecular size of a substrate increased, the affinity-immobilized trypsin showed higher clavage activity on insulin and BSA. This result seemed to indicate the streptavidin-biotin system allowed more steric flexibility of the immobilized trypsin in its interaction with a substrate molecule. To confirm this, we studied the molecular flexibility of immobilized trypsin using quartz crystal microbalance-dissipation. Self-assembled monolayers were formed on the Q-sensor surface by aminoalkanethiols, and gultaraldehyde was attached to the SAMs. Trypsin was immobilized in two ways: reductive amination(at pH 10.0) and the streptavidin-biotin system. The dissipation shift of the affinity-immobilized trypsin was $0.8{\times}10^{-6}$, whereas that of the covalently attached enzyme was almost zero. This result confirmed that the streptavidin-biotin system allowed higher molecular flexibility. These results suggested that the bioactivity of the immobilized enzyme be strongly dependent on its molecular flexibility.
Cadmium (Cd), a heavy metal found in the aquatic environment, accumulates in organisms through the food chain. In the study, we investigated the survival rates, measurement of body Cd levels, and expression analysis of the stress response genes (Heat shock protein 70: HSP70 and Heat shock protein 60: HSP60) and antioxidant enzyme Glutathione S-Transferases (GST) on benthic oligochaete worm Tubifex tubifex exposed three concentrations of Cd, to analyze the bioaccumulation and changes of stress gene expressions to exposure toxicity of the Cd-spiked sediment. Survival rates of T. tubifex exposed to the Cdspiked sediment were 93% at 0.4 mg kg-1 Cd, 96% at 1.87 mg kg-1 Cd, and 93% at 6.09 mg kg-1 Cd for 10 days. Cd concentration in the body of T. tubifex was higher than that in the sediment. After Cd exposures for 10 days, the body Cd levels were 18.4 mg kg-1, 13.06 mg kg-1, and 79.11 mg kg-1 at exposed three concentrations of Cd, respectively. Upregulation of HSP70 gene expression was observed at all concentrations of exposed Cd as a time-dependent manner, whereas transcriptional expression of the HSP60 gene increased as a timedependent manner in T. tubifex exposed to the relative high concentration (6.09 mg kg-1) of Cd. However, GST gene expression increased on day 1 at all concentrations after Cd exposures, and then downregulated until 10 days. These results indicate to ecotoxicological and molecular effects in benthic oligochaete worm T. tubifex to Cd-spiked sediment and provide the basic information for the utilization of environmental toxicity assessment using the T. tubifex as a aquatic pollution indicator species.
Objective : Allergic Inflammation is related with secretion of Cytokine. This study was performed to examine the effects of Woobangja on anti-allergic inflammation. Method : While macrophage 264.7cells was chosen as a normal group a control group was classified into three groups. One was stimulated with LPS. and another was pretreated with Woobangja for 1 hour. The third was pretreated with gydrocortisone for 1 hour. After the pretreatment, macrophage were incubated with lipopolysaccharide(LPS) 100 ng/ml for 12h and media collected and $TNF-{\alpha}$, IL-6, $IL-1{\beta}$, IL-10 concentration in supernatants were measured each by Enzyme linked immuno-sorbent assay. Woobangja were used $50\;{\mu}g/ml$, $100\;{\mu}g/ml$, $250\;{\mu}g/ml$, $500\;{\mu}g/ml$, 1 mg/ml. Hydrocortisones were used respectively $10^{-8}\;M$,$10^{-7}\;M$,$10^{-6}\;M$,$10^{-5}\;M$,$10^{-4}\;M$. Results : Woobangja showed inhibitory effect on $TNF-{\alpha}$ by LPS-stimulated macrophage 264.7. The inhibitory effect was most significant in 1mg/ml(p<0.01), and has increased according to the number of doses. Woobangja also showed inhibitory effect on IL-10 by LPS-stimulated macrophasg 264.7. The inhibitory effect was most significant in $100\;{\mu}g/ml$, and was not in a dose-dependent manner as Hydrocortisone group. Woobangja and Hydrocortison showed contrary effect on $IL-1{\beta}$ in al five concentration(p<0.01), and at the lowest concentration ($50\;{\mu}g/ml$) the level of $IL-1{\beta}$ was the lowest. On the other hand hydrocortison was observed to have inhibitory effect on $IL-1{\beta}$ in all five concentration(p<0.01). IL-6 was inhibited by hydrocortison in a roughly dose-dependent manner, but was not inhibited by Woobangja. On the contrary Woobangja obviously increased the expression of $IL-1{\beta}$ in all five concentration(p<0.01), but it was not related with concentrations. Conclusion : 1. Woobangja does significantly inhibit the expression of $TNF-{\alpha}$ by LPS-stimulated macrophage 264.7. 2. Woobangja does significantly increse the expression of IL-6 by LPS-stimulated macrophage 264.7. 3. Woobangja does significantly increse the expression of $IL-1{\beta}$ by LPS-stimulated macrophage 264.7. 4. Woobangja does significantly inhibit the expression of IL-10 by LPS-stimulated macrophage 264.7. 5. Woobangja is observer to have anti-allergic inflammatory effect through inhibiting inflammatory cytokine.
The present research was undertaken to investigate the activity of ligninolytic enzymes and the decolorization capability of some dyes with Irpex zonatus BN2, isolated from nature and identified. For the assay of enzyme activities, the isolate did not produce lignin peroxidase (LiP) and veratryl alcohol oxidase (VAO), but laccase and manganese dependent peroxidase (MnP). While the activity for MnP was low $(61.6\;nmol/mg{\cdot}protein)$, its laccase activity was very high $(1185.9\;nmol/mg{\cdot}protein)$. Moreover, laccase had appeared earlier than MnP. When the isolate was incubated with each dye for 10 days, the decolorization rates of azo dyes, such as orange II, orange G, tropaeolin O and congo red were 98.0%, 97.4%, 99.0% and 95.3%, respectively. In case of heterocyclic dyes, eosin Y, toludine blue, methyl blue and azur B were 97.4 %, 98.7%, 99.9% and 94.0% respectively. Finally the results of triphenylmethane dye such as basic fuchsin, malachite green and crystal violet were 98.5%, 95.7% and 99.4%, respectively. The results suggest that laccase of Irpex zonatus BN2 should be played an important role in the decolorization of the dyes.
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