• Journal of Plant Biotechnology
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• v.43 no.1
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• pp.37-48
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• 2016

The transcriptional profiles of 'Tamnara' grapevine (Vitis labruscana L.) to Rhizobium vitis were determined using 12,000 gene oligonucleotide microarray chips constructed with 6,776 unigenes based on the EST sequencing. Among them, 95 clones were up-regulated more than three times and 90 were down-regulated more than 5-times in the R. vitis-inoculated grapevines relative to the control vines. Treatment of salicylic acid showed that 337 clones were upregulated and 52 clones were down regulated in grapevines. Microarray analysis, reverse transcription-polymer chain reaction, and slot blot hybridization analysis revealed that 5, 14, and 64 clones were up-regulated and 10, 12, and 61 clones were down-regulated in wounded, salicylic acid-treated, and R. vitis-inoculated 'Tamnara' grapevine leaves, respectively. The expression patterns of ${\beta}$-1,3-glucanase, proline-rich protein, and lipoxygenase genes of 'Tamnara' moderately resistant to R. vitis were similar to those of resistant 'Concord' and 'Delaware' grapevines. However, chalcone synthase genes in 'Tamnara' grapevines showed similar expression patterns to susceptible grapevines 'Neomuscat' and 'Rizamat'. Further expression studies with various clones for each gene should be conducted to elucidate their roles in resistant responses against pathogens or other stimuli in grapevines. These results could provide better resources for understanding the mechanism of defense responses against crown gall disease and clues for identifying new genes that may play a role in defense against R. vitis in grapevines.

• Journal of Yeungnam Medical Science
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• v.34 no.1
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• pp.43-54
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• 2017

Background: Extracellular sulfatases (Sulfs), sulfatase 1 (Sulf1) and sulfatase 2 (Sulf2), play a pivotal role in cell signaling by remodeling the 6-O-sulfation of heparan sulfate proteoglycans on the cell surface. The present study examined the effects of Sulfs on angiotensin II (Ang II)-induced hypertensive mediator expression and vascular smooth muscle cells (VSMCs) proliferation in spontaneously hypertensive rats (SHR). Methods: Ang II receptors, 12-lipoxygenase (12-LO), and endothelin-1 (ET-1) messenger RNA (mRNA) expressions in SHR VSMCs were analyzed by real-time polymerase chain reaction and Western blotting. VSMCs proliferation was determined by [$^3H$]-thymidine incorporation. Results: Basal Sulfs mRNAs expression and enzyme activity were elevated in SHR VSMCs. However, Sulfs had no effect on the basal or Ang II-induced 12-LO and ET-1 mRNA expression in SHR VSMCs. The inhibition of Ang II-induced 12-LO and ET-1 expression by blockade of the Ang II type 2 receptor ($AT_2\;R$) pathway was not observed in Sulf1 siRNA-transfected SHR VSMCs. However, Sulf2 did not affect the action of $AT_2\;R$ inhibitor on Ang II-induced 12-LO and ET-1 expression in SHR VSMCs. The down-regulation of Sulf1 induced a reduction of $AT_2\;R$ mRNA expression in SHR VSMCs. In addition, the inhibition of Ang II-induced VSMCs proliferation by blockade of the $AT_2\;R$ pathway was mediated by Sulf1 in SHR VSMCs. Conclusion: These findings suggest that extracellular sulfatase Sulf1 plays a modulatory role in the $AT_2\;R$ pathway that leads to an Ang II-induced hypertensive effects in SHR VSMCs.

• Korean Journal of Food Science and Technology
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• v.39 no.3
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• pp.348-352
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• 2007

Inflammation is a complex process resulting from a variety of mechanisms. Combined inhibition of the activities of enzymes involved in the process may therefore be considered more important in anti-inflammatory property of plant extracts than any single contribution. In this study, the inhibitory effects of the ethanol extracts of thirty plant foods on the activities of secretory phospholipase $A_{2}$ ($sPLA_{2}$), cyclooxygenase-1 (COX-1), cyclooxygenase-2 (COX-2), and 12-lipoxygenase (12-LOX) were examined. Several legumes, mungbean sprout and some leaf vegetables inhibited the activity of $sPLA_2$, upstream enzyme of inflammation pathway. Only soybean sprout and mungbean sprout significantly inhibited 12-LOX activity. Although most of extracts inhibited the activities of both COX-1 and COX-2, water dropwort and amaranth showed selectivity for the inhibition of COX-2 over COX-1. Especially, mungbean showed anti-inflammatory property at both upstream and downstream of inflammation pathway with relatively low $IC_{50}$ values for $sPLA_{2}$ and COX-2 enzymes. Mungbean sprout exhibited inhibitory effects on all enzymes related to early and late inflammation and soybean sprout suppressed 12-LOX and COX-2 simultaneously, although the activities of these plants were showed at relatively high concentration. Therefore, mungbean, mungbean sprout, and soybean sprout appear to exhibit anti-inflammatory effects by combined inhibition of inflammatory enzymes.

• Journal of Nutrition and Health
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• v.56 no.6
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• pp.589-601
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• 2023

Purpose: Baicalein, a natural flavone found in herbs, exhibits diverse biological activities. Nonalcoholic steatohepatitis (NASH) is an irreversible condition often associated with a poor prognosis. This study aimed to evaluate the effects of baicalein on the development of NASH in mice. Methods: Male C57BL/6J mice were randomly divided into four groups. Three groups were fed a methionine-choline-deficient (MCD) diet to induce NASH and were simultaneously treated with baicalein (at doses of 50 and 100 mg/kg) or vehicle only (sodium carboxymethylcellulose) through oral gavage for 4 weeks. The control group was fed a methionine-choline-sufficient (MCS) diet without the administration of baicalein. Results: The baicalein treatment significantly reduced serum levels of alanine aminotransferase and aspartate aminotransferase, suggestive of reduced liver damage. Histological analysis revealed a marked decrease in nonalcoholic fatty liver activity scores induced by the MCD diet in the mice. Similarly, baicalein treatment at both doses significantly attenuated the degree of hepatic fibrosis, as examined by Sirius red staining, and hepatocellular death, as examined by the terminal deoxynucleotidyl transferase dUTP nick end labeling assay. Baicalein treatment attenuated MCD-diet-induced lipid peroxidation, as evidenced by lower levels of hepatic malondialdehyde and 4-hydroxynonenal, demonstrating a reduction in oxidative stress resulting from lipid peroxidation. Moreover, baicalein treatment suppressed hepatic protein levels of 12-lipoxygenase (12-Lox) induced by the MCD diet. In contrast, baicalein enhanced the activities of antioxidant enzymes such as superoxide dismutase, catalase, and glutathione peroxidase. Additionally, baicalein treatment significantly reduced hepatic non-heme iron concentrations and hepatic ferritin protein levels in mice fed an MCD diet. Conclusion: To summarize, baicalein treatment suppresses hepatic lipid peroxidation, 12-Lox expression, and iron accumulation, all of which are associated with the attenuation of NASH progression.

• The Korean Journal of Physiology and Pharmacology
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• v.2 no.6
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• pp.779-786
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• 1998

A reversed-phase high-performance liquid chromatogrphy (RP-HPLC) has been developed to analyze the metabolites of arachidonic acid based on the specificities of ultraviolet absorption of these various metabolites and is sensitive to the nanogram level. This procedure makes it possible to extract complex mixtures of eicosanoids efficiently with a single step and to analyze them simultaneously by RP-HPLC from biological samples using octadesylsilyl silica extraction column and $PGB_2$ as an internal standard. The cyclooxygenase products {prostaglandin $(PG)D_2,\;PGE_1,\;PGE_2,\;PGF_{1{\alpha}},\;PGF{2{\alpha}},\;6-keto-PGF_{1{\alpha}},$ and thromboxane $B_2(TXB_2)}$ and lipid peroxidation product, isoprostanes, of arachidonic acid were monitored by one isocratic HPLC system at 195 nm wavelength. The lipoxygenase products ${leukotriene(LT)B_4,\;LTC_4,\;LTD_4,$ and 5-hydroxyeicosatetraenoic acid (5-HETE), 12-HETE, 15-HETE} were measured by another isocratic HPLC system at 280 nm for LTs and 235 nm for HETEs. This method provides a simple and reliable way to extract and assess quantitatively the final arachidonic acid metabolites.

• Nutritional Sciences
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• v.9 no.3
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• pp.152-158
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• 2006

Helicobacter pylori (H. pylori) infection rapidly stimulated either COX-2 or 5-LOX and released arachidonic acid metabolites that have been considered as pivotal mediators in H. pylori-induced inflammatory responses. To determine whether red ginseng extract (RGE) can suppress the biosynthesis of 5(S)-hydroxyeicosatetraenoic acids (HETE), a precursor metabolite of leukotrienes B4 (LTB4) in H. pylori-provoked inflammatory responses in gastric epithelial cells, the biosynthesis of monohydroxy fatty acids was measured using radioactive arachidonic acid and validated by RP-HPLC using non-radioactive AA as substrate in AGS cells cocultured with H. pylori (ATCC 43504) with or without pretreatment of RGE. Among three known major HETEs, H. pylori infection specifically induced the biosynthesis of $^{14}C-5(S)-HETE$ rather than the complex of $^{14}C-15S-/^{14}C-12(S)-HETE$ from $^{14}C-AA$, concomitantly obtained by HPLC(p<0.01). RGE, 1 to $100{\mu}g/ml$, selectively suppressed H. pylori-stimulated $^{14}C-5(S)-HETE$ production implying the attenuation of 5-lipoxygenase activity, of which was similar to known LOX inhibitor NDGA $(10{\mu}M)$ (p<0.01). However, the amount of 5(S)-HETE was significantly reduced by higher dose of RGE $(100{\mu}g/ml)$ (p<0.05). These results indicated that LOX pathway might be one of principle pathogenic mechanisms of H. pylori and red ginseng could be a nutraceutical against H. pylori infection through inhibiting action of LOX activity.

• Journal of the Korean Society of Food Science and Nutrition
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• v.26 no.3
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• pp.376-382
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• 1997

Antioxidative characteristics of doenjang(fermented soybeans paste) phenolics on the lipid oxidation systems were studied by the determination of the oxidative related activity including lipoxygenase (LOX) inhibition, metal chelating and free radical scavengning of doenjang phenolics. Manlikong variety containing the highest amounts of phenolic compounds among the soybean variety, was used for doenjang processing. Doenjang was prepared by the series of processes including soaking for overnight, cooking for 1hr at 12Lb, first fermentation (3 days at 30$\pm$2$^{\circ}C$) for the preparation of meju(soybean koji) after inoculation of Asp. oryzae, and further fermentation(60 days at 30$\pm$2$^{\circ}C$) for the ripening after addition of salt 13% to meju. In order to investigate the antioxidative activity of phenolics in doenjang, the doenjang phenolics was extracted with methanol form freeze dried defatted doenjang. Antioxidative effects of methanol extract on linoleic acid oxidation system were observed by the significantly decreased levels of peroxide and conjugated diene formation. In addition, methanol extract resulted in the inhibition of LOX activity. and also, metal(FeCl$_3$) chelation and free radical scavengning activities were increased with increasing concentration of methanol extract.

• Korean Journal of Weed Science
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• v.12 no.2
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• pp.89-101
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• 1992

In this study, various physiological and biochemical experiments were conducted to know whether the selectivity between rice and barnyardgrass treated with bleaching herbicides containing diphenyl ether compounds was also partly based on their basic physiological proterties such as peroxidation ability, membrane stability or antioxidant system. It seemed to be partly based on the differences of their physiological characteristics that barnyardgrass was commonly more susceptible to most of the bleaching herbicides than rice. The scenescence of intact leaf segment was more rapid in barnyardgrass than in rice, indicating that barnyardgrass is weak at early stage. Also pigment metabolic ability, antioxidant enzyme activities(peroxidase, catalase, superoxide dismutase, glutathione reductase) and antioxidant content (tocopherol, ascorbic acid, glutathione, carotenoids) were lower in barnyardgrass on the basic of fresh weight. However, lipoxygenase activity and the content of unsaturated fatty acid which is vulnerable to oxygen radicals were higher in barnyardgrass, suggesting that barnyardgrass seedling bave a properties easy to be peroxidized. The differences of PPIX (protoporphyrin IX) or carotenoid content, which are the primary substances inducing herbicide activity, were not related to the selectivity between rice and barnyardgrass.

• Animal cells and systems
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• v.1 no.1
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• pp.81-86
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• 1997

We studied the mechanism of arachidonic acid on the secretion of a neurotransmitter in rat pheochromocytoma PC12 cells. Arachidonic acid inhibited the 70 mM $K^+$-induced secretion of norepinephrine. Arachidonic acid also inhibited the 70 mM $K^+$-induced $Ca^{2+}$ mobilization which is due to the opening of the voltage-sensitive $Ca^{2+}$ channels (VSCC). Both the half maximal inhibitory concentration ($IC_{50}$) of the norepinephrine secretion and VSCC coincided at 30 uM. The major oxidized metabolites of arachidonic acid, prostaglandins did not mimic the inhibitory effect of arachidonic acid. Nordihydroguaiaretic acid (NDGA) and indomethacin which are inhibitors of lipoxygenase and cyclooxygenase, respectively, did not block the inhibitory effect of arachidonic acid. The results suggest that arachidonic acid serves as a signal itself, not in the form of metabolites. The pretreatment of various $K^+$ channel blockers such as 4-aminopyridine, tetraethylarnmonium, glipizide, or glibenclamide also did not show any effect on the inhibitory effect of arachidonic acid. Through these results we suggest that arachidonic acid regulates VSCC directly and affects the secretion of neurotransmitters.

• Journal of the Korean Society of Food Science and Nutrition
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• v.42 no.3
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• pp.369-377
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• 2013

In order to increase the utilization of sweet potato leaves and stalks as much as roots, it is necessary to study their beneficial potential. In this study, the antioxidant, antiallergic and anti-inflammatory effects of sweet potato leaves and stalks were evaluated by measuring total polyphenol and flavonoid contents, DPPH radical scavenging effects, the reducing power and inhibition effects on xanthine oxidase (XO), 5-lipoxygenase (LOX), and cyclo-oxygenase (COX)-2 activities. Blanched sweet potato leaves (SL), raw whole purple stalks (ST) and peeled stalks (PST) were freeze-dried and extracted with 95% ethanol. Total polyphenol content was highest in SL (11.03 mg/g), followed by ST (0.87 mg/g), and PST (0.37 mg/g). Total flavonoid content was highest for SL (9.01 mg/g), followed by ST (0.50 mg/g) and PST (0.25 mg/g). The $IC_{50}$ for DPPH radical scavenging effects was highest for SL ($43.6{\mu}g/mL$), followed by ST ($308.4{\mu}g/mL$) and PST ($1,631.3{\mu}g/mL$). The reducing power was highest for SL ($59.72{\mu}g$ ascorbic acid eq./mL), followed by ST ($12.56{\mu}g$ ascorbic acid eq./mL) and PST ($2.18{\mu}g$ ascorbic acid eq./mL) with $1,000{\mu}g/mL$ of ethanol extract. The inhibition rate on XO activity was highest for SL (13.06%), followed by ST (5.05%) and PST (0.0%) at $250{\mu}g/mL$ extract treatment. The inhibition rate on COX-2 activity was highest for SL (55.34%), followed by ST (2.18%) and PST (0.0%) at $250{\mu}g/mL$ extract treatment. The inhibition rate on 5-LOX activity was highest for SL (91.16%), followed by ST (33.38%) and PST (14.93%) at $50{\mu}g/mL$ treatment. Taken together, sweet potato leaves showed high antioxidative, anti-allergic and anti-inflammatory activities, especially with very strong inhibition effects on 5-LOX activity. These beneficial effects of sweet potato leaves might be mainly caused by the high content of polyphenols and flavonoids.