Background: We previously demonstrated remarkable differences in the expression of IL-8/CXCL8 in aortic tissues and vascular smooth muscle cells (VSMC) from spontaneously hypertensive rats (SHR) compared to VSMC from normotensive Wistar-Kyoto rats (WKY). In the present study, we investigated the direct effect of IL-8/CXCL8 on expression of 12-lipoxygenase (LO), a hypertensive modulator, in SHR VSMC. Methods: Cultured aortic VSMC from SHR and WKY were used. Expression of 12-LO mRNA was determined by real-time polymerase chain reaction. Phosphorlyation of ERK1/2 and production of 12-LO and angiotensin II subtype 1 ($AT_1$) receptor were assessed by Western blots. IL-8/CXCL8-stimulated DNA synthesis was determined by measuring incorporation of [$^3H$]-thymidine. And effect of IL-8/CXCL8 on vascular tone was determined by phenylephrine-induced contraction of thoracic aortic rings. Results: Treatment with IL-8/CXCL8 greatly increased 12-LO mRNA expression and protein production compared to treatment with angiotensin II. IL-8/CXCL8 also increased the expression of the $AT_1$ receptor. The increase in 12-LO induced by IL-8/CXCL8 was inhibited by treatment with an $AT_1$ receptor antagonist. The induction of 12-LO mRNA production and the proliferation of SHR VSMC by IL-8/CXCL8 was mediated by the ERK pathway. The proliferation of SHR VSMC and the vascular contraction in the thoracic aortic ring, both of which were induced by IL-8/CXCL8, were inhibited by baicalein, a 12-LO inhibitor. Conclusion: These results suggest that the potential role of IL-8/CXCL8 in hypertensive processes is likely mediated through the 12-LO pathway.
Yoo, Sungjae;Han, Shanshu;Park, Young Shin;Lee, Jang-Hern;Oh, Uhtaek;Hwang, Sun Wook
Molecules and Cells
/
v.27
no.4
/
pp.417-422
/
2009
Lipoxygenase (LO) metabolites are generated in inflamed tissues. However, it is unclear whether the inhibition of the LO activity regulates the expression of c-Fos protein, a pain marker in the spinal cord. Here we used a carrageenan-induced inflammation model to examine the role of LO in the development of c-Fos expression. Intradermally injected carrageenan caused elevated number of cells exhibiting Fos-like immunoreactivity (Fos-LI) in the spinal dorsal horn, and decreased the thermal and mechanical threshold in Hargreaves and von Frey tests. Pretreatment with an inhibitor of phospholipase A2, that generates the LO substrate, prior to the carrageenan injection significantly reduced the number of Fos-(+) cells. A general LO inhibitor NDGA, a 5-LO inhibitor AA-861 and a 12-LO inhibitor baicalein also exhibited the similar effects. Moreover, the LO inhibitors suppressed carrageenan-induced thermal and mechanical hyperalgesic behaviors, which inidcates that the changes in Fos expression correlates with those in the nociceptive behaviors in the inflamed rats. LO products are endogenous TRPV1 activators and pretreatment with BCTC, a TRPV1 antagonist inhibited the thermal but not the mechanical hypersensitivity. Overall, our results from the Fos-LI and behavior tests suggest that LO products released from inflamed tissues contribute to nociception during carrageenan-induced inflammation, indicating that the LO pathway is a possible target for modulating inflammatory pain.
Son Min-Jung;Moon Tae-Chul;Lee Eun-Kyung;Son Kun-Ho;Kim Hyun-Pyo;Kang Sam-Sik;Son Jong-Keun;Lee Seung-Ho;Chang Hyeun-Wook
Archives of Pharmacal Research
/
v.29
no.4
/
pp.282-286
/
2006
Ochnaflavone is a medicinal herbal product isolated from Lonicera japonica that inhibits cyclooxygenase-2 (COX-2) dependent phases of prostaglandin $D_2(PGD_2)$ generation in bone marrow-derived mast cells (BMMC) in a concentration-dependent manner with $IC_{50}$ values of $0.6{\mu}M$. Western blotting probed with specific anti-COX-2 antibodies showed that the decrease in quantity of the $PGD_2$ product was accompanied by a decrease in the COX-2 protein level. In addition, this compound consistently inhibited the production of leukotriene $C_4 (LTC_4)$ in a dose dependent manner, with an $IC_{50}$ value of $6.56 {\mu}M$. These results demonstrate that ochnaflavone has a dual cyclooxygenase-2/5-lipoxygenase inhibitory activity. Furthermore, this compound strongly inhibited degranulation reaction in a dose dependent manner, with an $IC_{50}$ value of $3.01{\mu}M$. Therefore, this compound might provide a basis for novel anti-inflammatory drugs.
Kim, Dong-Yung;Kim, Jun-Hyung;Lee, Jae-Chul;Won, Moo-Ho;Yang, Se-Ran;Kim, Hyoung-Chun;Wie, Myung-Bok
Toxicological Research
/
v.35
no.1
/
pp.83-91
/
2019
Nanoparticles (NPs) have been recognized as both useful tools and potentially toxic materials in various industrial and medicinal fields. Previously, we found that zinc oxide (ZnO) NPs that are neurotoxic to human dopaminergic neuroblastoma SH-SY5Y cells are mediated by lipoxygenase (LOX), not cyclooxygenase-2 (COX-2). Here, we examined whether human bone marrow-derived mesenchymal stem cells (MSCs), which are different from neuroblastoma cells, might exhibit COX-2- and/or LOX-dependent cytotoxicity of ZnO NPs. Additionally, changes in annexin V expression, caspase-3/7 activity, and mitochondrial membrane potential (MMP) induced by ZnO NPs and ZnO were compared at 12 hr and 24 hr after exposure using flow cytometry. Cytotoxicity was measured based on lactate dehydrogenase activity and confirmed by trypan blue staining. Rescue studies were executed using zinc or iron chelators. ZnO NPs and ZnO showed similar dose-dependent and significant cytotoxic effects at concentrations ${\geq}15{\mu}g/mL$, in accordance with annexin V expression, caspase-3/7 activity, and MMP results. Human MSCs exhibited both COX-2 and LOX-mediated cytotoxicity after exposure to ZnO NPs, which was different from human neuroblastoma cells. Zinc and iron chelators significantly attenuated ZnO NPs-induced toxicity. Conclusively, these results suggest that ZnO NPs exhibit both COX-2- and LOX-mediated apoptosis by the participation of mitochondrial dysfunction in human MSC cultures.
Leukotriene B4 (LTB4) is a lipid mediator of inflammation that is generated from arachidonic acid via the 5-lipoxygenase pathway. Previous studies have reported that the receptors of LTB4, BLT1, and BLT2 play mediatory roles in the allergic airway inflammation induced by ovalbumin (OVA). However, considering that house dust mites (HDMs) are the most prevalent allergen and well-known risk factor for asthmatic allergies, we are interested in elucidating the contributory roles of BLT1/2 in HDM-induced allergic airway inflammation. Our aim in this study was to investigate whether BLT1/2 play any roles in HDM-induced allergic airway inflammation. In this study, we observed that the levels of ligands for BLT1/2 [LTB4 and 12(S)-HETE (12(S)-hydroxyeicosatetraenoic acid)] were significantly increased in bronchoalveolar lavage fluid (BALF) after HDM challenge. Blockade of BLT1 or BLT2 as well as of 5-lipoxygenase (5-LO) or 12-lipoxygenase (12-LO) markedly suppressed the production of TH2 cytokines (IL-4, IL-5, and IL-13) and alleviated lung inflammation and mucus secretion in an HDM-induced eosinophilic airway-inflammation mouse model. Together, these results indicate that the 5-/12-LO-BLT1/2 cascade plays a role in HDM-induced airway inflammation by mediating the production of TH2 cytokines. Our findings suggest that BLT1/2 may be a potential therapeutic target for patients with HDM-induced allergic asthma.
Sarath Ly;Hyeon Su Oh;Se Yeong Kim;Jeong Hwan Lee;Jong Il Chung
KOREAN JOURNAL OF CROP SCIENCE
/
v.68
no.3
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pp.114-120
/
2023
Soybean is the one of the most important crops for providing quality vegetable protein to umans and livestock. Soybean cultivars with a brown seed coat have a wide range of antioxidant benefits because of the flavonoid components. However, they also contain lectin, 7S α′ subunit, lipoxygenase, and Kunitz trypsin inhibitor (KTI) proteins that can be allergenic and digestive inhibitors and reduce processing aptitude. Genetic removal of these four proteins is necessary in soybean breeding. Therefore, this study was conducted to select a new line with brown seed coat, green cotyledon, and tetra-null genotype (lecgy1lox1lox2lox3ti) for lectin, 7S α′ subunit, lipoxygenase, and KTI proteins in the mature seed. Five germplasms were used to create breeding population. From a total of 58 F2 plants, F2 plants with lele genotype were selected using a DNA marker, and F3 seeds with a brown seed coat, green cotyledon, and the absence of 7S α′ subunit protein were selected. Three lines (S1, S2, and S3) were developed. Genetic absence of lectin, 7S α′ subunit, lipoxygenase, and KTI proteins was confirmed in F6 seeds of the three lines, which had a brown seed coat, green cotyledons, and a white hilum. The 100 seed weights of the three lines were 26.4-30.9 g, which were lower than 36 g of the check cultivar - 'Chungja#3'. The new S2 line with 30.9 g hundred seed weight can be used as a parent to improve colored soybean cultivars without antinutritional factors such as lectin, 7S α′ subunit, lipoxygenase, and KTI proteins.
An investigation was conducted to ascertain the basic information on characteristics of embryonic tissue development among Korean Ginseng, American ginseng, $F_{1}$ hybrids and $BC_{1}F_{1}(F_{1}$ pollen back cross to $BC_{1}=Korean\;Ginseng)seeds$. The specific activities of lipoxygenase (LOX) and the protein assays were made during the above embryonic growth period at 5 or 25 degree C. The results are summarized as follows: The fresh weights(mg) of germinating seeds were significantly different among species. Ginseng seeds were stratified at $5^{\circ}C\;for\;60\;days$ the ratios of embryo/endosperms in $P.g.,\;P.q\;or\;F_{1}\;were\;about\;90{\%}$. The ratio was in $BC_{1}F_{1},\;79.2{\%}$ during the same period. The ratios of embryo/endosperms of seeds of P.g. or P.q. germinated at $5^{\circ}C$ showed rather higher values as compared to that at $25^{\circ}C$ LOX specific activity the highest in the $F_{1}$ and decreased in the order of P.g., P.q., and $BC_{1}F_{1}$. The highest LOX specific activity was shown at 80 days after sowing, followed by 70 days sowing, the least LOX activity was shown at 60 days.
Journal of the Society of Cosmetic Scientists of Korea
/
v.30
no.1
/
pp.23-28
/
2004
The aim of this study is to assess the anti-irritation activities of mung bean (Phaseolus aureus) extract against various irritants used in cosmetics. For its antidotal activity, mung bean has been used as a medicinal or cosmetic material since ancient times. However, there have been few reports describing the biological activities of these beans and no comprehensive surveys of the constituents. We obtained an ethanolic extract of mung bean and isolated the major constituents, such as vitexin and isovitexin. And we previously reported that the mung bean extract containing vitexin and isovitexin had excellent antioxidant and anti-inflammatory activities. To investigate the mechanisms of anti-inflammatory activity of mung bean extract, we examined the inhibitory effects on histamine release from rat peritoneal mast cells and lipoxygenase activity. Mung bean extract inhibited histamine release in a concentration dependent manner but showed no inhibitory activity in the 5-lipoxygenase assay. And, clinical studies were conducted to evaluate the anti-irritation effects of mung bean extract against various irritants used in cosmetics such as lactic acid, retinol, and preservatives. When 2.0% of mung bean extract was applied to cosmetic formulae containing each of irritants, it revealed considerable anti-irritation efficacy. Our results of the human patch test with 20 volunteers showed that this extract reduced skin irritations caused by 5.0% lactic acid, 4000 IU retinol, and 1.0% preservative mixture by about 60%, 30%, and 50% respectively. The stinging potential test for assessing subjective irritation also showed that the extract reduced the unpleasant sensations by about 50∼30%. Finally, we performed a double-blind usage test with 30 subjects to compare formulae containing mung bean extract with placebo. From the results of questionnaires for 4 weeks of use, we confirmed the excellent anti-irritation effect of mung bean extract. Conclusively, we could discover new material that had anti-irritation effects and apply this mung bean extract to the final cosmetic products successfully.
Park, Sung-Kyu;Son, Na-Young;Kim, Mi-Jung;Chung, Ill-Min
Proceedings of the Korean Society of Crop Science Conference
/
2017.06a
/
pp.264-264
/
2017
Legumes are good sources of various nutrients. Among legumes, soybean and its flour are accessible foods to consumers. However, in case of soybean flour, there is a disadvantage of easily going rancid. Thus, this study aimed to investigate how the quality of soybean flour changes during storage according to the packaging materials and storage temperatures. The raw and roasted soybean flours were packed in two types of packaging, polyethylene (PE) and polypropylene (PP) film bags respectively, and stored at three different storage temperatures (4, 20, and $45^{\circ}C$) for 1 year. The acid value, conjugated diene value, peroxide value, p-anisidine value, thiobarbituric acid (TBA) value, lipoxygenase activity, and fatty acid content of raw and roasted soybean flours were measured at the point of starting storing, and after 1, 2, 4, 8, and 12 weeks of storage. The acid value of soybean flour was increased for 4 weeks and thereafter significantly decreased (p < 0.0001). The conjugated diene value was significantly increased after 4 weeks storage at $45^{\circ}C$ with PE and PP film bags (p < 0.0001). The peroxide value had no changes during 4 weeks storage at $45^{\circ}C$ with PE and PP film bags, and then those was dramatically increased after 8 weeks (p < 0.0001). The p-anisidine values in all of storage conditions were decreased after 4 weeks. Lipoxygenase activity was decreased at 12 weeks storage at $20^{\circ}C$ and $45^{\circ}C$ with PP film bags. Acid value had positive correlations with p-anisidine value (r = 0.30 and p < 0.0001) and lipoxygenase activity (r = 0.36 and p < 0.0001), and had negative correlations with conjugated diene value (r = -0.45 and p < 0.0001) and peroxide value (r = -0.25 and p < 0.001). Conjugated diene value had a high positive correlation with peroxide value (r = 0.76 and p < 0.0001), but that had a negative correlation with lipoxygenase activity (r = -0.51 and p < 0.0001). Peroxide value had negative correlations with p-anisidine value (r = -0.20 and p < 0.01) and TBA value (r = -0.15 and p < 0.05). The degree of reduction in fatty acid content of raw soybean flour was higher than the roasted soybean flour during the storage. Total fatty acid content had positive correlations with acid value (r = 0.45 and p < 0.0001) and p-anisidine value (r = 0.58 and p < 0.0001), but had a weak negative correlation with conjugated diene value (r = -0.19 and p < 0.01). This study showed how the rancidity of the raw and roasted soybean flours progressed during storage. Thus, our findings can be used as base data to do a further study of finding and developing more stable storage conditions of the soybean flour.
Proceedings of the Korean Society of Crop Science Conference
/
2017.06a
/
pp.69-69
/
2017
Due to the lack in storage facility, considerable amount of rice is stocked in the open air, which causes increased stale flavor and deteriorates palatability and merchantable quality. Lipoxygenase-3(LOX-3) is involved in the production of volatile constituents in stored rice, and the development of stale flavor is delayed in LOX-3 null rice. LOX activity in the rice grain is localized in bran fraction and the tropical Japonica cultivar 'Dawdam' was reported that the absence of LOX-3 is inherited as a simple recessive trait. Also, it was reported that the peroxidation of unsaturated fatty acids occurs at lower levels in the 'Dawdam' bran fraction during storage than in rice varieties with LOX-3. This study was conducted to develop LOX-3 null rice lines using 'Dawdam' and investigate changes of physicochemical properties of the lines stored at different storage temperatures and periods. So we analyzed texture, toyo glossiness value, germination rate and lipoxygenase activity of 15 LOX-3 null rice lines on the condition of which rough rice had been stored at different temperatures (high temperature condition at $35^{\circ}C$ and low temperature condition at $15^{\circ}C$ for 4months. Hardness and stickiness of the lines tendered to be increased when it was stored at high temperature and adhesiveness, springiness, cohesiveness and chewiness was not considerably different according to storage temperatures and periods. The germination rate of HR29062-B-98-2-1-B among LOX-3 null rice lines was higher than another lines, 99.3, 94.0% after 4months stored at low temperature and high temperature, respectively. The lipoxygenase activity was 3,304, 1,601unit/mg protein after 4months stored at low temperature and high temperatures, respectively. So, it is thought that this line will be useful to breed rice varieties with high storability after tested on agricultural traits.
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