• Korean Journal of Food Science and Technology
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• v.54 no.2
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• pp.147-154
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• 2022

The Mongolian lingonberry and blueberry are two essential food sources found in Mongolia. This study investigated the antioxidant and anti-inflammatory effects of methanol extracts from Mongolian lingonberry (LBE) and blueberry (BBE). Compared to the LBE, the BBE showed higher total phenolic, flavonoid, and anthocyanin contents, as well as antioxidant capacities. The LBE and BBE inhibited the mRNA expression of pro-inflammatory genes, including tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β), and cyclooxygenase (COX-2) in lipopolysaccharide-stimulated RAW 264.7 macrophage cells. In addition, the LBE and BBE inhibited NADPH oxidase-2 (Nox2) mRNA expression, indicating that they have cellular antioxidant capacities. Anthocyanin derivatives of the LBE and BBE were analyzed using LC-QTOF/MS. Six anthocyanins were identified in the BBE, while one was detected in the LBE. Our findings demonstrate that the anthocyanin-rich LBE and BBE could be used as functional food sources in Mongolia.

• The Korean Journal of Physiology and Pharmacology
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• v.26 no.4
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• pp.263-275
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• 2022

There is a paucity of detailed data related to the effect of senescence on the mitochondrial antioxidant capacity and redox state of senescent human cells. Activities of TCA cycle enzymes, respiratory chain complexes, hydrogen peroxide (H₂O₂), superoxide anions (SA), lipid peroxides (LPO), protein carbonyl content (PCC), thioredoxin reductase 2 (TrxR2), superoxide dismutase 2 (SOD2), glutathione peroxidase 1 (GPx1), glutathione reductase (GR), reduced glutathione (GSH), and oxidized glutathione (GSSG), along with levels of nicotinamide cofactors and ATP content were measured in young and senescent human foreskin fibroblasts. Primary and senescent cultures were biochemically identified by monitoring the augmented cellular activities of key glycolytic enzymes including phosphofructokinase, lactate dehydrogenase, and glycogen phosphorylase, and accumulation of H₂O₂, SA, LPO, PCC, and GSSG. Citrate synthase, aconitase, α-ketoglutarate dehydrogenase, succinate dehydrogenase, malate dehydrogenase, isocitrate dehydrogenase, and complex I-III, II-III, and IV activities were significantly diminished in P25 and P35 cells compared to P5 cells. This was accompanied by significant accumulation of mitochondrial H₂O₂, SA, LPO, and PCC, along with increased transcriptional and enzymatic activities of TrxR2, SOD2, GPx1, and GR. Notably, the GSH/GSSG ratio was significantly reduced whereas NAD⁺/NADH and NADP⁺/NADPH ratios were significantly elevated. Metabolic exhaustion was also evident in senescent cells underscored by the severely diminished ATP/ADP ratio. Profound oxidative stress may contribute, at least in part, to senescence pointing at a potential protective role of antioxidants in aging-associated disease.

• Journal of Ginseng Research
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• v.46 no.4
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• pp.515-525
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• 2022

Background: The incidence of ischemic cerebrovascular disease is increasing in recent years and has been one of the leading causes of neurological dysfunction and death. Ginsenoside Rg1 has been found to protect against neuronal damage in many neurodegenerative diseases. However, the effect and mechanism by which Rg1 protects against cerebral ischemia-reperfusion injury (CIRI) are not fully understood. Here, we report the neuroprotective effects of Rg1 treatment on CIRI and its possible mechanisms in mice. Methods: A bilateral common carotid artery ligation was used to establish a chronic CIRI model in mice. HT22 cells were treated with Rg1 after OGD/R to study its effect on [Ca²⁺]_i. The open-field test and poleclimbing experiment were used to detect behavioral injury. The laser speckle blood flowmeter was used to measure brain blood flow. The Nissl and H&E staining were used to examine the neuronal damage. The Western blotting was used to examine MAP2, PSD95, Tau, p-Tau, NOX2, PLC, p-PLC, CN, NFAT1, and NLRP1 expression. Calcium imaging was used to test the level of [Ca²⁺]_i. Results: Rg1 treatment significantly improved cerebral blood flow, locomotion, and limb coordination, reduced ROS production, increased MAP2 and PSD95 expression, and decreased p-Tau, NOX2, p-PLC, CN, NFAT1, and NLRP1 expression. Calcium imaging results showed that Rg1 could inhibit calcium overload and resist the imbalance of calcium homeostasis after OGD/R in HT22 cells. Conclusion: Rg1 plays a neuroprotective role in attenuating CIRI by inhibiting oxidative stress, calcium overload, and neuroinflammation.

• The Korea Journal of Herbology
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• v.35 no.1
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• pp.57-68
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• 2020

Objective : The aim of present study was to clarify the effect of Areca Semen and Toosendan Fructus Mixture (AT-mix) on chronic reflux esophagitis (CRE) in rats. Methods : The antioxidant activity of AT-mix was measured through DPPH and ABTS radical scavenging activities in vitro. CRE was induced in SD rats (5 weeks, male) by ligating the border forestomach and granular portion with 2-0 silk and the duodenum near the pyloric portion was covered with 2-mm wide piece of 18-Fr Nélaton catheter. And then rats were treated AT-mix 200 mg/kg one daily for 14 days. The anti-oxidant and inflammatory protein levels were evaluated using western blotting. Results : Gross lesion of esophageal mucosa after AT-mix treatment showed a superior enhancement compared with that of CRE control rats. AT-mix treatment strongly reduced both DPPH and ABTS radical scavenging activities (DPPH, IC₅₀ 8.15±0.14 ㎍/mL; ABTS, IC₅₀ 24.69±0.03 ㎍/mL, repspectively). Levels of the NADPH oxidase subunit including NOX4 and p22^phox increased in CRE control rats. Otherwise, AT-mix treatment significantly reduced. The activation of Nuclear factor-erythroid 2-related factor 2 (Nrf2) led to significantly the up-regulation of HO-1. The inhibition of IκBα phosphorylation led to NF-κB inactivation. Subsequently, NF-κB inactivation significantly induced the decrease of COX-2, iNOS, TNF-α, and IL-6 protein expressions. Conclusion : Taken together, these results suggest that AT-mix treatment can attenuate the esophageal mucosal ulcer though inhibiting NF-κB pathway and enhancing Nrf2/HO-1 pathway. Thus, the additional mechanism study about AT-mix would need for the development as a safe herbal therapy for CRE.

• The Korea Journal of Herbology
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• v.37 no.5
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• pp.63-74
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• 2022

Objective : Gastritis refers to an inflammatory disease of the gastric mucosa. Alcohol is one of the main aggression factors, causing bleeding and inflammation in the gastric mucosa and it is known to not only increase lipid peroxide levels, but also deplete key antioxidant factors. The purpose of this study was to determine the effect of Uncariae Ramulus et Uncus water extract (URW) in alcohol-induced gastritis. Methods : The total polyphenol and flavonoid contents of URW were confirmed through an in vitro experiment. Also, 2,2-diphenyl-1-picrylhydrazyl (DPPH) and 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid (ABTS) radical scavenging activity and ferric reducing antioxidant power (FRAP) activity were confirmed. For in vivo experiments, mice were divided into 4 groups (n=8). Also, 1 hr after oral administration of each drug, 50% ethanol was orally administered to induce gastritis. Results : As a result of in vitro experiments, URW showed excellent antioxidant activity. In alcohol-induced gastritis, URW alleviated the damage to the gastric mucosa caused by alcohol. Also, URW decreased reactive oxygen species (ROS) and malondialdehyde (MDA) levels in serum and gastric tissues, and significantly decreased the expression of NADPH oxidases in gastric tissues. In addition, it significantly modulated the nuclear factor erythroid-derived 2-related factor 2 (Nrf2) and nuclear factor-𝜅B p65 (NF-𝜅B) pathways as well as significantly increased the expression of anti-inflammatory proteins. Conclusions : These results suggest that URW not only reduces oxidative stress through excellent antioxidant activity but also relieves gastric mucosal inflammation as a regulator of Nrf2 and NF-𝜅B pathways.

• Asian-Australasian Journal of Animal Sciences
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• v.14 no.5
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• pp.615-623
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• 2001

A study of mammary function in relation to glucose metabolism of first lactation Bali cows on grass-legume diets was carried out using 12 primiparous cows (initial BW $263.79{\pm}21.66kg$) for 16 weeks starting immediately post calving. The animals were randomly allocated into 4 dietary treatment groups R1, R2, R3 and R4, receiving from the last 2 months of pregnancy onwards, rations based on a mixture of locally available grass and legume feed ad libitum. On a DM basis R1 contained 70% elephant grass (PP, Penicetum purpureum) plus 30% Gliricidia sepia leaves (GS), R2 was 30% PP plus 25% GS supplemented with 55% Hibiscus tilliacius leaves (HT, defaunating effect), R3 and R4 were 22.5% PP+41.25% GS+11.25% HT+25% concentrate, with R4 supplemented with zinc-diacetate. TDN, CP and zinc contents of the diets were 58.2%, 12.05% and 18.3 mg/kg respectively for R1, 65.05%, 16.9% and 25.6 mg/kg respectively for R2, 66.03%, 16.71% and 29.02 mg/kg respectively for R3 and 66.03%, 16.71% and 60.47 mg/kg respectively for R4. Milk production and body weights were monitored, an energy and protein balance trial conducted, overall glucose kinetics parameters assessed, mammary blood flow (MBF) and metabolite arteriovenous differences (${\Delta}AVs$) measured to get uptake data and mammary performance relationships. Parameters of glucose kinetics at peak lactation or during dry condition were not affected by ration quality. Glucose pool size, space of distribution and flux increased by 61.77, 62.26 and 82.08%, respectively, during lactation compared to the dry period. Mean glucose flux of lactating Bali cows was $5.52mg/min.kgBW^{0.807}$ which resembles the range of values of temperate dairy cows. Calculation showed that glucose requirements for maintenance, milk lactose and fat-glycerol synthesis, and the formation of NADPH reached 461.69 g for a yield of 1 kg/d or equal to 320.62 mg/min, which was less than the average glucose flux of lactating Bali cows of 481.35 mg/min. Mammary blood flow (MBF) values ranged from 56 to 83 l/h for the different treatments and the ratio MBF per kg milk produced improved from av. 1540 l/kg for R1 to av. 967 l/kg for R4 treated cows. Mammary glucose uptake ranged from 6.27 to 12.03 g/h or 120 to 140 g/kg milk. Glucose uptake was mass-wise 2 to 4 times the amount secreted as lactose, which indicated values less than the calculated mammary glucose needs and that little lactose was synthesized. The excess glucose taken-up was used for other metabolic processes. Linear relationships between metabolite ${\Delta}AVs$ and arterial blood plasma concentration [A] showed that in Bali cows triglycerides (TG), phenylalanine (Phe) and tyrosine (Tyr) have high coefficients of determination, i.e. 0.77, 0.81 and 0.69, respectively. For glucose, the relationship is quadratic with an $R^2$ value of 0.49. It was concluded that lactose synthesis was inadequate, which led to a speculation that milk yield could be improved by increased lactose synthesis.

• 한국균학회소식:학술대회논문집
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• 2014.05a
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• pp.27-28
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• 2014

Beauveria bassiana (Cordycipitaceae, Hypocreales, Ascomycota) is an anamorphic fungus having a potential to be used as a biological control agent because it parasitizes a wide range of arthropod hosts including termites, aphids, beetles and many other insects. A number of bioactive secondary metabolites (SMs) have been isolated from B. bassiana and functionally verified. Among them, beauvericin and bassianolide are cyclic depsipeptides with antibiotic and insecticidal effects belonging to the enniatin family. Non-ribosomal peptide synthetases (NRPSs) play a crucial role in the synthesis of these secondary metabolites. NRPSs are modularly organized multienzyme complexes in which each module is responsible for the elongation of proteinogenic and non-protein amino acids, as well as carboxyl and hydroxyacids. A minimum of three domains are necessary for one NRPS elongation module: an adenylation (A) domain for substrate recognition and activation; a tholation (T) domain that tethers the growing peptide chain and the incoming aminoacyl unit; and a condensation (C) domain to catalyze peptide bond formation. Some of the optional domains include epimerization (E), heterocyclization (Cy) and oxidation (Ox) domains, which may modify the enzyme-bound precursors or intermediates. In the present study, we analyzed genomes of B. bassiana and its allied species in Hypocreales to verify the distribution of NRPS-encoding genes involving biosynthesis of beauvericin and bassianolide, and to unveil the evolutionary processes of the gene clusters. Initially, we retrieved completely or partially assembled genomic sequences of fungal species belonging to Hypocreales from public databases. SM biosynthesizing genes were predicted from the selected genomes using antiSMASH program. Adenylation (A) domains were extracted from the predicted NRPS, NRPS-like and NRPS-PKS hybrid genes, and used them to construct a phylogenetic tree. Based on the preliminary results of SM biosynthetic gene prediction in B. bassiana, we analyzed the conserved gene orders of beauvericin and bassianolide biosynthetic gene clusters among the hypocrealean fungi. Reciprocal best blast hit (RBH) approach was performed to identify the regions orthologous to the biosynthetic gene cluster in the selected fungal genomes. A clear recombination pattern was recognized in the inferred A-domain tree in which A-domains in the 1st and 2nd modules of beauvericin and bassianolide synthetases were grouped in CYCLO and EAS clades, respectively, suggesting that two modules of each synthetase have evolved independently. In addition, inferred topologies were congruent with the species phylogeny of Cordycipitaceae, indicating that the gene fusion event have occurred before the species divergence. Beauvericin and bassianolide synthetases turned out to possess identical domain organization as C-A-T-C-A-NM-T-T-C. We also predicted precursors of beauvericin and bassianolide synthetases based on the extracted signature residues in A-domain core motifs. The result showed that the A-domains in the 1st module of both synthetases select D-2-hydroxyisovalerate (D-Hiv), while A-domains in the 2nd modules specifically activate L-phenylalanine (Phe) in beauvericin synthetase and leucine (Leu) in bassianolide synthetase. antiSMASH ver. 2.0 predicted 15 genes in the beauvericin biosynthetic gene cluster of the B. bassiana genome dispersed across a total length of approximately 50kb. The beauvericin biosynthetic gene cluster contains beauvericin synthetase as well as kivr gene encoding NADPH-dependent ketoisovalerate reductase which is necessary to convert 2-ketoisovalarate to D-Hiv and a gene encoding a putative Gal4-like transcriptional regulator. Our syntenic comparison showed that species in Cordycipitaceae have almost conserved beauvericin biosynthetic gene cluster although the gene order and direction were sometimes variable. It is intriguing that there is no region orthologous to beauvericin synthetase gene in Cordyceps militaris genome. It is likely that beauvericin synthetase was present in common ancestor of Cordycipitaceae but selective gene loss has occurred in several species including C. militaris. Putative bassianolide biosynthetic gene cluster consisted of 16 genes including bassianolide synthetase, cytochrome P450 monooxygenase, and putative Gal4-like transcriptional regulator genes. Our synteny analysis found that only B. bassiana possessed a bassianolide synthetase gene among the studied fungi. This result is consistent with the groupings in A-domain tree in which bassianolide synthetase gene found in B. bassiana was not grouped with NRPS genes predicted in other species. We hypothesized that bassianolide biosynthesizing cluster genes in B. bassiana are possibly acquired by horizontal gene transfer (HGT) from distantly related fungi. The present study showed that B. bassiana is the only species capable of producing both beauvericin and bassianolide. This property led to B. bassiana infect multiple hosts and to be a potential biological control agent against agricultural pests.

• The Korean Journal of Pharmacology
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• v.16 no.2 s.27
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• pp.45-53
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• 1980

Lipid peroxidation in vitro has been identified as a basic deteriorative reaction in cellular mechanism of aging processes, such as air pollution oxidant damage to cell and to the lung, chlorinated hydrocarbon hepatotoxicity. Many experimental evidences were reported by several investigators that lipid peroxidation could be one of the principle causes for the hepatotoxicity produced by $CCl_4$. It is now reasonably established that $CCl_4$ is activated to a free radical in vivo, that lipid peroxidation occurs very quickly in microsomes prepared from damaged livers, that the peroxidation is associated with loss of enzyme activity of microsomes, and that various antioxidants can protect animals against the hepatotoxic effect of $CCl_4$. Recent studies have drawn attention to some other feature of microsomal lipid peroxidation. Incubation of liver microsomes in the presence of NADPH has led to a loss of cytochrome $P_{450}$. However, the presence of an antioxidant prevented lipid peroxidation and preserved cytochrome $P_{450}$. Decrease of cytochrome $P_{450}$ in microsomes under in vitro incubation can be enhanced by $CCl_4 and these changes were parallel to a loss of microsomal polyunsaturated fatty acid and formation of malonaldehyde. The primary purpose of this experiment was to study the effect of riboflavin tetrabutylate on lipid peroxidation, specially, the relationship between lipid peroxidation and drug metabolizing enzyme system which is located in smooth endoplasmic recticulum as well as the effect of ritoflavin tetrabutylate on drug metabolizing enzyme system of animal treated with $CCl_4$. Albino rats were used for experimental animal. In order to induce drug metabolizing enzyme system, phenobarbital was injected intraperitoneally. $CCl_$ and riboflavin tetrabutylate were given intraperitoneally as solution in olive oil. Microsomal fraction was isolated from liver of animals and TBA value as well as the activity of drug metabolizing enzyme were measured in the microsomal fractions. The results are summerized as following. 1) The secobarbital induced sleeping time of $CCl_4$ treated rat was about 2 times longer than that of the control group. However, the pretreatment with riboflavin tetrabutylate inhibited completely the lengthened sleeping time due to $CCl_4$ treatment. Furthermore TBA value was significantly increased in $CCl_4$ treated rat in comparison to control group tut the increase of TBA value was prevented by the pretreatment with riboflavin tetrabutylate. On the other hand, the activity of hepatic drug metabolizing enzyme was decreased in $CCl_4$ group, however, the pretreatment with riboflavin tetrabutylate also prevented the decrease of the enzyme activity caused by $CCl_4$. 2) The effect of riboflavin tetrabutylate on TBA value and the activity of drug metabolizing enzyme in vitro was similar to in vivo results. Incubation of liver microsome from rat in the presence of $CCl_4$, $Fe^{++}$, or ascorbic acid has led to the marked increase of TBA value, however, the addition of riboflavin tetrabutylate in incubation mixture prevented significantly the increase of TBA value, suggesting the inhibition of lipid peroxidation. In accordance with TBA value, the activity of drug metabolizing enzyme was inhibited in the presence of $CCl_4$, $Fe^{++}$, ascorbic acid but the addition of riboflavin tetrabutylate protected the loss of the enzyme activity in microsome under in vitro incubation.

• The Korean Journal of Pharmacology
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• v.21 no.1
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• pp.62-77
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• 1985

In tile Iron-catalyzed Haber-Weiss reaction to produce OH., the requirement for $O^{-}_{2}{\cdot}$ is only to reduce $Fe^{+++}$. Possibly, the role of $O^{-}_{2}{\cdot}$ can be replaced by other reducing agents. Ascorbate is one of them in biological system. In the present study, the ability of ascorbate to produce $OH{\cdot}$ in the presence of $Fe^{++}$ and $H_2O_2$ was investigated by observing the degradation of hyaluronic acid and ethylene production from methional. Ascorbate stimulated the degradation of hyaluronic by $Fe^{++}$ and $H_2O_2$. That was confirmed by both viscosity change and gel-permeation chromatographic analysis. The observed degradation was almost completely prevented by catalase and $OH{\cdot}$ scavengers. In support of the above results, ascorbate enhanced the prouction of ethylene from methional in the presence of $Fe^{++}$ and $H_2O_2$. Other reducing agents (cysteine, glutathione, NADH and NADPH) showed similar activities to ascorbate in the degradation of hyaluronic acid and ethylene production. But no stimulatory effects were observed with their oxidized forms such as NAD and NADP. Thus, it appears that reduction of the metal ion was needed for $OH{\cdot}$ production. Among the metal ions tested, $Fe^{++}$ showed most potent catalytic action in the production of $OH{\cdot}$ The results obtained support that ascorbate can substitute $O^{-}_{2}{\cdot}$ in the metal-catalyzed reactions, particularly with $Fe^{++}$ by which $OH{\cdot}$ is produced with $H_2O_2$. The significance of the ascorbate-dependent production of $OH{\cdot}$ was considered with respect to possible role of ascorbate in the damage of inflamed joints.

• Food Science and Preservation
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• v.19 no.1
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• pp.123-130
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• 2012

In this study, 80% ethanolic extracts of tartary and common buckwheats were assessed for their total phenol content, total flavonoids content, antioxidant activity (DPPH, ABTS radical scavenging activity and reducing power), and anti-adipogenic effects. Our results show that total phenol contents of 80% ethanolic extract from tartary and common buckwheats were $17.35{\pm}0.41$ and $8.20{\pm}0.28\;{\mu}g$ GAE/g, respectively. Antioxidant activities of 80% ethanolic extract from tartary buckwheat were significantly higher than that of common buckwheat extract (p<0.05). During adipocyte differentiation, 80% ethanolic extracts of tartary and common buckwheat significantly inhibited lipid accumulation compared to control cells. We further evaluated the effect of buckwheat extracts on the changes of key gene expression associated with 3T3-L1 adipogenesis and ROS production. Tartary buckwheat extract was more suppressed the mRNA expressions ($PPAR{\gamma}$ and aP2) than that of common buckwheat extract. Moreover, tartary buckwheat inhibited the mRNA expression of both NOX4 (NADPH oxidase 4) and G6PDH (glucose-6-phosphate dehydrogenase). These results indicate that anti-adipogenesis effect of tartary buckwheat can be attributed to phenolic compound that may potentially inhibit ROS production.