The present study performed the isolation of cytosolic ascorbate peroxidase (APX) isozymes and analyzed the pattern of their activity development and also investigated the change in some other enzyme activities related to the ascorbate-glutathione pathway from the senescing wheat leaves. The aim of this work is to examine the possibility that in the cytoplasm of wheat leaves the ascorbate-glutathione pathway p!ays a significant role in relation to leaf senescence involving an $H_2O_2$ accumulation and then to show the effect of benzyladenine (BA) on that pathway. During the leaf senescence characterized by increases in ChI breakdown and H202 accumulation under the 4-day dark incubation of matured leaf segments; i) no significant increase of total cytosolic APX was observed, ii) a dehydroascorbate reductase (DHAR) activity was decreased rapidly, iii) a slight increase of glutathione reductase (GR) activity occurred. In the BA-treated leaves; however, i) the total activity of APX increased conspicuously, ii) the decrease of DHAR activity was relatively inhibited, iii) the GR activity increase was more enhanced, and iv) the decrease of ascorbate content and the increase of H202 content were retarded as compared with those of control leaves. Three isozymes of cytosolic APX were found by using a native-electrophoretic gel in senescing wheat leaves and two of them occurred with major activity. In the developmental patterns of cytosolic APX isozymes, only two isozyme bands ("a" and "b") appeared with almost constant activity through 4 days of incubation in the control leaves, while one additional weak isozyme band ("c") and a little increase of "b" isozyme activity were detected in the BA-treated leaves. EspeciaUy, the development of "a" isozyme activity increased remarkably compared with that of control leaves. The increased capacity for peroxide scavenging due to the enhanced activity of all 3 enzymes (APX, DHAR, GR) participating in the ascorbate-glutathione pathway in BA-treated leaves suggested that this pathway might playa significant role in the processes related to the wheat leaf senescence.scence.
Namkoong, Seung;Jang, Seon-A;Sohn, Eun-Hwa;Bak, Jong Phil;Sohn, Eunsoo;Koo, Hyun Jung;Yoon, Weon-Jong;Kwon, Jung-Eun;Jeong, Yong Joon;Meng, Xue;Han, Hyo-Sang;Kang, Se Chan
Korean Journal of Plant Resources
/
v.28
no.2
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pp.145-152
/
2015
The present study aimed to investigate comparative anti-inflammatory effects of Litsea japonica fruit and leaf extract considering the balance of safety and efficacy. Dose response studies were performed to determine the inhibitory effects of 70% EtOH extract of leaf (L70%) on the pro-inflammatory enzymes expression, COX-2/PGE2 and NO/iNOS, and pro-inflammatory cytokines production, IL-$1{\beta}$, IL-6, and TNF-$\alpha$ in lipopolysaccharide (LPS)-stimulated RAW 264.7 cells. We also examined comparative effects of 30 and 70% EtOH extract of fruits (F30% and F70%) at low concentration ($10{\mu}g/ml$ ) in the same conditions. L70% at 50 and $100{\mu}g/ml$ showed inhibitory effects on almost all the inflammatory mediators we examined except for COX-2 regulation, but there were no effects at $10{\mu}g/ml$. Since $100{\mu}g/ml$ of L70% have 18.2% cytotoxicity, we compared the effects of fruit extract, F30% and F70% at $10{\mu}g/ml$ on the regulation of NO/iNOS, PGE2, IL-$1{\beta}$, IL-6, and TNF-$\alpha$ and obtained that fruit extacts are more efficacious and safe than leaf. This study suggests that the 30% EtOH fraction of L. japonica fruit could be a good candidate for development as a functional food supplement in the prevention of inflammatory disorders.
Lee, Young-Kyung;Kim, Chul Hwan;Jeong, Dae Won;Lee, Ki Won;Oh, Young Taek;Kim, Jeong Il;Jeong, Jin-Woo
Korean Journal of Plant Resources
/
v.35
no.5
/
pp.565-573
/
2022
Gingival inflammation is one of the main causes that can be related to various periodontal diseases. Human gingival fibroblast (HGF) is the major constituent in periodontal connective tissue and secretes various inflammatory mediators, such as nitric oxide (NO) and prostaglandin E2 (PGE2), upon lipopolysaccharide stimulation. This study is aimed at investigating the anti-inflammatory and antioxidative activities of Lotus Root extract (LRE) in Porphyromonas gingivalis derived lipopolysaccharide (LPS-PG)-stimulated HGF-1 cells. The concentration of NO and PGE2, as well as their responsible enzymes, inducible NO synthase (iNOS), and cyclooxygenase-2 (COX-2), was analyzed by Griess reaction, ELISA, and western blot analysis. LPS-PG sharply elevated the production and protein expression of inflammatory mediators, which were significantly attenuated by LRE treatment in a dose-dependent manner. LRE treatment also suppressed activation of Toll-like receptor 4 (TLR4)/myeloid differentiation primary response gene 88 (MyD88) and nuclear factor-κB (NF-κB) in LPS-PG-stimulated HGF-1 cells. In addition, one of phase II enzyme, NAD(P)H quinone dehydrogenase (NQO)-1, and its transcription factor, Nuclear factor erythroid 2-related factor 2 (Nrf2), were significantly induced by LRE treatment. Consequently, these results suggest that LRE ameliorates LPS-PG-induced inflammatory responses by attenuating TLR4/MyD88-mediated NF-κB, and activating NQO-1/Nrf2 antioxidant response element signaling pathways in HGF-1 cells.
Poultry are exposed to extremely high levels of oxidative stress as a consequence of the excessive production of reactive oxygen species (ROS) induced by endogenous and exogenous stressors, such as high-stocking densities, thermal stress, environmental and feed contamination, along with factors associated with intensive breeding systems. Oxidative stress promotes lipid peroxidation, DNA damage, and inflammation, which can have detrimental effects on the health of birds. During the course of evolution, birds have developed antioxidant defense mechanisms that contribute to maintaining homeostasis when exposed to endogenous and exogenous stressors. The primary antioxidant defense systems are enzymatic and non-enzymatic in nature and play roles in protecting cells from ROS attack. Recently, plant flavonoids, which have been established to reduce oxidative stress, have been attracting considerable attention as potential feed additives. Flavonoids are a group of polyphenolic compounds that can be stabilized by binding structural compounds with ROS, and can promote the elimination of ROS by inducing the expression of antioxidant enzymes. However, although flavonoids can contribute to reducing lipid peroxidation and thereby enhance the antioxidant capacity of birds, they have low solubility in the gastrointestinal tract, and consequently, it is necessary to develop a delivery technology that can facilitate the effect intestinal absorption of these compounds. Furthermore, it is important to determine the dietary levels of flavonoids by assessing the exact antioxidant effects in the gastrointestinal tract wherein the concentrations of dietary flavonoids are highest. It is also necessary to examine the expression of transcriptional factors and vitagenes associated with the efficient antioxidant effects induced by flavonoids. It is anticipated that the application of flavonoids as natural antioxidants will become a particularly important field in the poultry industry.
Su Jeong Kim;Hwang Bae Sohn;A Hyun Park;Jong Nam Lee;Su Hyoung Park;Jung Hwan Nam;Do Yeon Kim;Dong Chil Chang;Yul Ho Kim
Korean Journal of Plant Resources
/
v.36
no.5
/
pp.435-445
/
2023
The aim of this study was to explore the effects of vinegar extract from seed of common buckwheat (Fagopyrum esculentum Moench) and seed of tartary buckwheat (F. tataricum Gaertner) on acute ethanol-induced hangover in Sprague-Dawley rats. Vinegar extract from buckwheat is rich choline, quercetin and its glycoside, rutin known as flavonoid antioxidants. The test extract containing buckwheat was proven to alleviate hangovers through a significant reduction in the concentration of alcohol and acetaldehyde in the context of an alcohol-induced hangover model. Hepatic alcohol dehydrogenase (ADH) and acetaldehyde dehydrogenase (ALDH) activities were significantly higher in buckwheat vinegar-treated rats than in ethanol-treated rats. Moreover, tartary buckwheat vinegar upregulated antioxidant enzyme such as superoxide dismutase and Catalase activities in liver tissues. These results suggest that buckwheat vinegar extract could alleviate ethanol-induced hangover symptoms by elevating activities related to hepatic ethanol-metabolizing enzymes against ethanol induced metabolites, and in particular, tartary buckwheat should be further developed to be a novel anti-hangover material.
The nitrate ($NO_3{^-}$) accumulation of hydroponically grown leafy vegetables may increase in the condition of a closed-type plant production system with low light intensity due to low activity of enzymes involved in nitrogen assimilation and the use of $NO_3-N$ as major nitrogen source. The objective of this study is to investigate the effects of light intensities, nutrient solution compositions and the time of nutrient solution removal before harvest on nitrate contents of hydroponically-grown lettuces in a closed plant production system. The reduction of nitrate contents in leafy lettuces 'Cheongchima' was higher in the treatments of 'TW' (nutrient solution removal) and '$(NH_4)_2CO_3$' (use of ammonium carbonate as nitrogen source) than those in other treatments, which significantly lowered fresh weight and leaf area of the plants. In the light intensity of $100{\mu}mol{\cdot}m^{-2}{\cdot}s^{-1}$, the nitrate content was effectively reduced without causing any growth retardation, by substitution of the nutrient solution composition that $NO_3-N$ was removed ('$NO_3-N$ removal' treatment) or the half strength of standard nutrient solution was applied ('1/2 S' treatment), for 7days before harvest. The effects of light intensity and the time of nutrient solution removal before harvest on growth and nitrate contents in leafy lettuces were investigated. The nitrate contents in leaves under the light condition of $300{\mu}mol{\cdot}m^{-2}{\cdot}s^{-1}$ before nutrient solution removal were lower than those of 100 or $200{\mu}mol{\cdot}m^{-2}{\cdot}s^{-1}$. The removal of nutrient solution for 7 days before harvest quickly reduced the amount of nitrates in leaves in all the light intensities with a greater degree under the $300{\mu}mol{\cdot}m^{-2}{\cdot}s^{-1}$ of light condition, while the 7 days-removal with both 200 and $300{\mu}mol{\cdot}m^{-2}{\cdot}s^{-1}$ of light conditions caused decrease in 16~31% of leaf area and 20~35% of fresh weight, compared to the 3 days-removal treatment. The nitrate contents were greatly reduced from 3,018 to 1,035 in $200{\mu}mol{\cdot}m^{-2}{\cdot}s^{-1}$, and 2,021 to 480 ppm in the light condition of $300{\mu}mol{\cdot}m^{-2}{\cdot}s^{-1}$, with the nutrient solution removal for 3 days before harvest, without causing any deterioration in growth and product quality. The vitamin C contents in leaves were higher in the treatment of nutrient solution removal for both 3 and 5 days before harvest with the light condition of $300{\mu}mol{\cdot}m^{-2}{\cdot}s^{-1}$ than those in the light condition of 100 or $200{\mu}mol{\cdot}m^{-2}{\cdot}s^{-1}$.
Antioxidative function of uncooked powdered food (Sangsik) was evaluated in rats consuming nutritionally unbalanced diet including 1% cholesterol, high proportion of animal lipids (lard : soybean oil : 8 . 2) , sub-optimal levels of vitamin and mineral mixture along with 0.5% ethanol in drinking water. The uncooked powdered food tested in the present study was a mixture composed of 42 kinds of plant foods (cereals, legumes, seaweeds, vegetables, and fruits) supplemented with vitamins and minerals, and dietary fiber. Control rats were fed the semi-purified diet based on the AIN-93G composition, and nutritionally unbalanced rats were divided into 3 groups, and fed one of the following diets with 0.5% ethanol in drinking water for 5 weeks : unbalanced control diet (UC) ,20% Sangsik powder supplemented diet (S20), and 40% Sangsik powder supplemented diet (S40) . Food efficiency ratio was significantly higher in rats fed S40 compared to the value for rats fed UC (p < 0.05). Hepatic level of thiobarbituric acid reactive substances (TBARS) was significantly lower in rats fed UC compared to that for control rats (p < 0.05) , and was not influenced by dietary supplementation of the Sangsik powder. Hepatic superoxide dismutase (SOD) activity was significantly higher in rats fed UC compared to that for control rats (p < 0.05) , and significantly reduced in rats fed S20 or S40 compared to the value for unbalanced control rats. Feeding unbalanced control diet significantly reduced the ratio of hepatic GSH-Px + catalase/SOD activities compared to the value for control rats, and this decrease in the ratio of antioxidant enzyme activities was reversed by adding the Sangsik powder to the diet at 20% (p <0.05) . Based on the results of antioxidant enzyme activities, feeding uncooked powdered diet appears to provide a favorable environment for body's antioxidative defense mechanism. Serum levels of Fe and Cu were significantly lower in rats fed the Sangsik powder supplemented diets compared to the value for unbalanced control rats (p < 0.05) , and levels of Se, Mn, and Zn were also tended to be decreased by dietary supplementation of the Sangsik powder. These results postulate the possibility that ingredients used in the uncooked powdered food may decrease the bioavailability of trace elements in rats.
In order to understand more clearly the integration between N-assmilation and C-metabolism in relation to N fertilization, a pot experiment with 5 different level of N fertilization(0, 5, 10, 25, 50 mM NO$_3$$_{[-10]}$ ) was conducted in Manchester, U.K. The peas (Pisum sativum L., cv. Early Onward) were sown in vermiculate (5 cm depth) and cultivated for 6 days under temperature controlled dark room conditions ($25^{\circ}C$). The plants received frequent irrigation with a nutrient solution: it was fertilized every 2 days, 3 times a day at 10h, 13h, 16h respectively. Elevated NO$_3$$^{[-10]}$ concentration, the activity levels of NR, NiR, total GS(crude extract), GS$_2$(plastid) in both root and shoot were increased and reached the peak in 5~25 mM, except NiR specific activity which increased its activity continually until 50 mM NO$_3$$^{[-10]}$ treatment. Total activities of GS (crude extract) in both root and shoot became higher than those of GS$_2$(Plastid), and the activity ratios of total GS in the crude extract and GS$_2$ in the plastids were 3.0 to 4.3 in root, but 3.2 to 10.6 in shoot. It was concluded that the reductants and A TP from OPPP itself should be enough to achieve the high rate of NR, NiR, GS$_1$, GS$_2$ in plant root and shoot for reduction or assimilation of nitrogen, but these enzyme activities might be inhibited by an excess of NO$_3$$^{[-10]}$ influx over the reduction capacity.
Kim, Seul-Ki;Hwang, Hyun-Jin;Kim, Jae-Deog;Ko, Eun-Hye;Choi, Jung-Sup;Kim, Jin-Seog
Korean Journal of Weed Science
/
v.32
no.2
/
pp.85-97
/
2012
To investigate the usefulness of freshwater alga Water-net (Hydrodictyon reticulatum, HR) as resources for production of fermentable sugars, the easiness of enzymatic saccharification was evaluated at first. When 6 plant materials (HR, Spirulina, Chlorella, Scenedesmus, Cladophora, Corn stover) were enzymatically hydrolyzed with 2% solid loading at the same condition, HR showed the highest ratio of saccharification based on glucose production. No milled HR was also completely saccharified at the amounts of optimal enzyme mixture. Glucose yield was not changed though the citrate buffer strength for saccharification was decreased from 0.1 M to 0.1 mM. Only about 10% yield reduction was observed compared to that of $120^{\circ}C$ treatment when HR was enzymatically hydrolyzed at room temperature. The saccharification was normally occurred at $37^{\circ}C$ and pH 6.5 which is general growth condition of fermentable microrganisms, suggesting that HR have a biomass characteristics applicable for the simultaneous saccharification and fermentation. The saccharification was occurred by more than 70~80% of one of the best condition although the supplied enzyme amounts was reduced to 1/10 volume. And the glucose yield by enzymatic hydrolysis was not decreased by 10% HR solid loading and began to decrease at more than 15% solid contents. Above these results show that HR is an interesting algal biomass which is relatively easy to be saccharified by hydrolyzing enzymes. In addition, HR is a flilamentous alga and very easy to be collected. Therefore, HR seems to be an useful and valuable resources in the economical production of fermentable sugars for manufacture of bio-chemical products.
In order to characterize the property of $Ca^{2+}$ transport in plant cell, microsomes were prepared from the roots of tomato and microsomal $^{45}Ca^{2+}$ uptake was measured. When 1 mM vanadate, a selective inhibitor of P-type ATPases, 50 mM $NO_3^-$, a specific inhibitor of vacuolar $H^{+}-ATPase$, and both of these inhibitors were treated, the microsomal $^{45}Ca^{2+}$ uptakes were inhibited by 20, 33 and 47%, respectively. The inhibitory effects of these two inhibitors were investigated by using a protonophore, gramicidin. When the chemical gradient of $H^{+}$ was relieved by gramicidin, the uptake was decreased by 30%, implying the presence of $Ca^{2+}/H^+$ antiporter in the microsomal membrane. In the $^{45}Ca^{2+}$ uptake experiment, the effect of gramicidin was independent of vanadate-induced inhibition. However, when the activity of vacuolar $H^{+}-ATPase$ was inhibited by $NO_3^-$, the effect of gramicidin was severely decreased. Meanwhile, thapsigargin, a specific antagonist of ER/SR-type $Ca^{2+}-ATPase$, inhibited the microsomal $^{45}Ca^{2+}$ uptake and the maximum inhibitory effect was obtained at $10\;{\mu}M$. The effect of thapsigargin was blocked by $NO_3^-$ and gramicidin, but not by vanadate. These results imply that vanadate directly inhibits the activity of $Ca^{2+}-ATPase$; however, $NO_3^-$ and thapsigargin block the activity of $Ca^{2+}/H^+$ antiporter by inhibiting the vacuolar $H^{+}-ATPase$. In conclusion, the microsomal $^{45}Ca^{2+}$ uptakes are mediated by two major enzymes, $Ca^{2+}-ATPase$ and $Ca^{2+}/H^+$ antiporter in tomato root tissue.
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