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Oxymatrine inhibits the pyroptosis in rat insulinoma cells by affecting nuclear factor kappa B and nuclear factor (erythroid-derived 2)-like 2 protein/heme oxygenase-1 pathways

  • Gao, Jingying;Xia, Lixia;Wei, Yuanyuan
    • The Korean Journal of Physiology and Pharmacology
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    • v.26 no.3
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    • pp.165-174
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    • 2022
  • As the mechanism underlying glucose metabolism regulation by oxymatrine is unclear, this study investigated the effects of oxymatrine on pyroptosis in INS-1 cells. Flow cytometry was employed to examine cell pyroptosis and reactive oxygen species (ROS) production. Cell pyroptosis was also investigated via transmission electron microscopy and lactate dehydrogenase (LDH) release. Protein levels were detected using western blotting and interleukin (IL)-1β and IL-18 secretion by enzyme-linked immunosorbent assay. The caspase-1 activity and DNA-binding activity of nuclear factor kappa B (NF-κB) and nuclear factor (erythroid-derived 2)-like 2 protein (Nrf2) were also assessed. In the high glucose and high fat-treated INS-1 cells (HG + PA), the caspase-1 activity and LDH content, as well as Nod-like receptor family pyrin domain containing 3, Gsdmd-N, caspase-1, apoptosis-associated speck-like protein containing a CARD, IL-1β, and IL-18 levels were increased. Moreover, P65 protein levels increased in the nucleus but decreased in the cytoplasm. Oxymatrine attenuated these effects and suppressed high glucose and high fat-induced ROS production. The increased levels of nuclear Nrf2 and heme oxygenase-1 (HO-1) in the HG + PA cells were further elevated after oxymatrine treatment, whereas cytoplasmic Nrf2 and Keleh-like ECH-associated protein levels decreased. Additionally, the elevated transcriptional activity of p65 in HG + PA cells was reduced by oxymatrine, whereas that of Nrf2 increased. The results indicate that the inhibition of pyroptosis in INS-1 cells by oxymatrine, a key factor in its glucose metabolism regulation, involves the suppression of the NF-κB pathway and activation of the Nrf2/HO-1 pathway.

Comparison of Physiological Activities of Radish Bud (Raphanus sativus L.) according to Extraction Solvent and Sprouting Period (추출용매 및 발아시기에 따른 무순 추출물의 생리활성 비교)

  • Han, Jin-Hee;Moon, Hye-Kyung;Chung, Shin-Kyo;Kang, Woo-Won
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.44 no.4
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    • pp.549-556
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    • 2015
  • This study extracted radish bud (Raphanus sativus L.) and investigated its nitrite scavenging activity, superoxide dismutases (SOD)-like activity, tyrosinase inhibition activity, xanthine oxidase inhibition activity, and angiotensin-converting-enzyme (ACE) inhibition activity according to extraction solvent and sprouting period. For nitrite scavenging activity, each extract recorded its highest level of 81.44~89.71% at pH 1.2. Radish bud extracts on sprouting days 4 and 8 showed greater scavenging activities than those on sprouting day 12 at pH 1.2 and pH 4.0. There were differences in scavenging activity according to extraction solvent based on water extract exhibiting improved scavenging activity. Ethanol extract recorded scavenging activity of 16.12% at pH 6.0, which was similar to those of ethanol and methanol radish bud extracts on sprouting day 12. SOD-like activity of radish bud extracts was in the range of 4.57~27.05%. For comparison purposes, SOD-like activity of L-ascorbic acid was 52.15%, which was higher than that of radish bud extracts. Acetone and methanol extracts showed high SOD-like activities on sprouting day 8. SOD-like activity of radish bud extracts on sprouting day 12 significantly decreased to 4.57~15.59%. Radish bud extracts recorded good tyrosinase inhibitory activities on sprouting 8 and 12, whereas methanol extracts recorded the greatest tyrosinase inhibitory activity at 62.65~84.89%. Radish bud extracts recorded xanthine oxidase inhibition activity of 21.26~29.52% on sprouting day 4, and acetone extracts showed the highest level of xanthine oxidase inhibition activity. Xanthine oxidase inhibitory activity tended to decrease with sprouting period compared early on. ACE inhibitory activity was in the range of 12.48~51.78% according to sprouting period and extraction solvent. Ethanol extracts on sprouting day 8 showed the highest ACE inhibitory activity of 51.78%. These results will hopefully contribute to research into the identification of materials and development of products for natural functional foods.

Antioxidative Activity of the Durian (Durio zibethinus) Extract (두리안 추출물의 항산화 활성)

  • Ji, Yun Sun;Chang, Jun Pok
    • Korean Journal of Medicinal Crop Science
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    • v.21 no.4
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    • pp.255-261
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    • 2013
  • The purpose of this study was to examined the antioxidant activities by water and 70% ethanol extract from durian (Durio zibethinus.) seed, sarcocarp and peel. Durian extract were studied for reducing sugar content, polyphenol content, superoxide dismutase (SOD) like activity, electron donating ability, nitrite scavenging ability, flavonoid content, hydroxy radical scavenging activity. Reducing sugar content were increased peel > sarcocarp > seed. Total polyphenol, flavonoid content, DPPH radical scavenging ability and SOD like activity were increased seed > peel > sarcocarp. Total polyphenol content was relatively high as $21.90{\pm}0.50mg/g$ in the ethanol extract of the seed. DPPH radical scavenging ability was relatively high as $62.08{\pm}2.63%$ in the water extract of the seed. Nitrite scavenging ability was no significant difference. Hydroxy radical scavenging activity was increased seed > peel > sarcocarp, was relatively high as $58.27{\pm}1.13%$ in the water extract of the seed.

Antioxidant activity of ethanol extracts from the root and bark of Ulmus davidiana

  • Sim, Ki Hyeon;Han, Young Sil
    • Korean Journal of Food Science and Technology
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    • v.51 no.2
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    • pp.133-140
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    • 2019
  • Antioxidant activities of Ulmus davidiana root and bark extracts were evaluated by various antioxidant tests, including DPPH radical-scavenging, nitric oxide-scavenging, superoxide anion radical-scavenging, and ABTS radical-scavenging assays, and superoxide dismutase (SOD)-like activity and reducing power analysis, along with the determination of total phenolic and flavonoid contents. Both the extracts showed strong antioxidant activities by these testing methods. Ulmus davidiana root extract possessed strong reducing power and nitric oxide-scavenging activity, and high scavenging activities against free radicals including the superoxide anion, and the ABTS and DPPH radicals, but a weaker scavenging activity of SOD. In contrast, the Ulmus davidiana bark extract exhibited a strong SOD-like activity, but all the other activities were weak. It was observed that the total phenolic and flavonoid contents of the Ulmus davidiana root extract were higher than those of the Ulmus davidiana bark extract.

Effects of Alkaline Phosphatase Activity on the Extract of Carthami Semen and Eucommiae Cortex in Human Osteoblast-like MG-63 Cell Line (홍화자와 두충 혼합 추출물이 MG-63 조골세포의 Alkaline Phosphatase 활성에 미치는 영향)

  • Sim, Jae-Geun;Lee, Jae-Hyeok;Yeo, Myeong Gu;Park, Jeong-Suk
    • Journal of the East Asian Society of Dietary Life
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    • v.23 no.1
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    • pp.39-43
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    • 2013
  • Carthamus tinctorius L. and Eucommia umoides Oliver are often used in traditional herbal medicines for reducing damage to the liver, kidney, bone and muscle. In the present study, we investigated cell viability and alkaline phosphatase activity in the human osteoblast-like MG-63 cell line with methanol extracts of Carthami Semen (CS) and Eucommiae Cortex (EC) alone or in a mixture (CS+EC). Osteoblast cell viability was evaluated using the MTS assay and alkaline phosphatase activity assays. The cell viability and alkaline phosphatase activity significantly increased in MG-63 osteoblast cells treated with the CS+EC mixture. These findings suggest the CS+EC mixture may have beneficial effects on bone health through the proliferation of osteoblast cells.

Protective Effect of Omega-3 of Polyunsaturated Fatty Acid Docosahexaenoic Acid on the Organic Mercury-Induced Cytotoxicity in Cultured NIH3T3 Fibroblasts

  • Ha, Dae-Ho;Lee, Jai-Kyoo
    • Biomedical Science Letters
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    • v.14 no.3
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    • pp.187-192
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    • 2008
  • To clerify the protective effect of omega-3 of polyunsaturated fatty acid docosahexaenoic acid (DHA) on the cytotoxicity induced by organic mercury in cultured NIH3T3 fibroblasts. The measurement of cell viability on ogranic mercury wad done by XTT assay after NIH3T3 fibroblasts were cultured with various concentrations of methyl mercuric chloride (MMC). And also, the effect of DHA on the MMC-mediated cytotoxicity was examined by cell viability, and antioxidant effect of DHA was also assessed by superoxide dismutase (SOD)-like activity and the lipid peroxidation activity in cultured NIH3T3 fibroblasts. In this study, MMC decreased cell viability and $XTT_{50}$ value was determined at $50{\mu}M$ of MMC in these culture. In the effect of DHA against the cytotoxicity induced by MMC, DHA significantly increased the cell viability damaged by MMC in cultured NIH3T3 fibroblasts. And also, DHA showed the antioxidant effect by showing the increase of SOD-like activity and the decrease of lipid peroxidation activity. From these results, it is suggested that organic mercury such as MMC has highly toxic effect on cultured NIH3T3 fibroblasts, and also, omega-3 of polyunsaturated fatty acid, DHA showed the protection on MMC-induced cytotoxicity and antioxidant effect.

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Antioxidant Activity and Anti-inflammatory Effects of Salix Koreensis Andersson Branches Extracts (버드나무(Salix Koreensis Andersson) 가지 추출물의 항산화 및 항염증 효과)

  • Kim, Mi-Hye
    • Journal of the Korean Society of Food Culture
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    • v.33 no.2
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    • pp.104-111
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    • 2018
  • This study aims to compare and analyze a willow tree (Salix Koreensis andersson) extract's antioxidant and anti-inflammatory activity by investigating its: total polyphenol, flavonoid content, SOD-like activity, DPPH vitality. the willow tree was induced with LPS to determine its active anti-inflammatory effects. as a result, the willow methanol extract showed a higher total polyphenol and flavonoid content than those of willow distilled water extract, but the willow distilled water extract showed a higher SOD than that of willow methanol extract. in its DPPH scavenging ability, the willow methanol extract's antioxidant activity was higher than that of the willow distilled water extract. the willow extract's measurements such as the production of NO, inflammatory cytokine ($TNF-{\alpha}$, IL-6 measurement) were significantly reduced as its concentration level went down. according to the research outcomes, when induced, he will extract's macrophage produces mediator-like substances such as NO and inflammatory cytokine that can be used to alleviate the inflammatory response. therefore, the willow tree proved to be a useful raw plant material for the products designed to combat inflammatory activities due to its natural antioxidant and anti-inflammatory response substances such as NO and cytokine.

Developmental Changes in Digestive Enzymes Activity of Black Rockfish Sebastes inermis (볼락, Sebastes inermis 자.치어의 성장에 따른 소화효소 활성 변화)

  • Gwak Woo-Seok;Park Dae-Won
    • Journal of Aquaculture
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    • v.19 no.2
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    • pp.125-132
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    • 2006
  • Black rockfish larvae and juveniles were reared for 95 days after parturition (DAP) in order to determine four enzyme activities (trypsin, pepsine-like enzyme, lipase, amylase) during ontogeny. Larvae were fed rotifers Brachionus plicatilis from 1 to 25 DAP, Artemia nauplii from 10 to 78 DAP and then gradually changed to pelleted feed from 30 DAP. Temperature was kept between $13.5{\sim}14.9^{\circ}C$. Trypsin and lipase activities were found in 2 DAP larvae ($7.0{\pm}1.5$ unit and $4.5{\pm}1.4$ unit, $mean{\pm}SD$, respectively). The evolution of both enzymes activities showed a profile marked by drastic increases between postflexion and juvenile stage. There is an increment on specific trypsin activity at 10 DAP, corresponding with the beginning of Artemia feeding. Pepsin-like enzyme activity was found at 11 DAP and increased drastically from 56 DAP, cor-responding with the initiation of juvenile stage. Amylase activity was also found at 11 DAP and maintained at a low level up to 38 DAP followed by a drastic increase from 39 DAP to 50 DAP. Considering our results of both trypsin and pepsin-like enzyme activities, it might be concluded that higher somatic growth of Sebastes inermis could be possible with the initiation juvenile of stage and the early juvenile stage is a suit-able period for feeding an artificial diet for fish.

ROLE OF NF${\kappa}B$ IN TOLL-LIKE RECEPTOR 9-MEDIATED MATRIX METALLOPROTEINASE-9 EXPRESSION (Toll-like receptor 9-매개에 의한 matrix metalloproteinase-9 발현에서 NF${\kappa}B$의 역할)

  • Lee, Sang-Hoon;Chin, Byung-Rho;Baek, Suk-Hwan
    • Journal of the Korean Association of Oral and Maxillofacial Surgeons
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    • v.33 no.6
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    • pp.636-642
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    • 2007
  • Background: CpG DNA plays an important role in immune cell function. This study examined whether the temporal control of toll-like receptor (TLR)9 by CpG DNA can regulate the expression of matrix metalloproteinase-9(MMP-9). Methods and materials: Macrophages were cultured in the presence of 10% FBS. For the various MMP genes analysis, RT-PCR and real-time PCR were performed. In addition, zymography assay performed for the MMP activity. The phosphorylation assay did for the ERK1/2 and NF${\kappa}B$ activation, and luciferase promoter assay was for the NF${\kappa}B$ activity. Results: CpG DNA induced the mRNA expression of MMP-2, MMP-9, and MMP-13, but not of MMP-7, MMP-8, and MMP-12, in a time-dependent manner. Especially, the mRNA expression of MMP-9 was strongly induced by CpG DNA using real-time RT-PCR. The TLR9 inhibitor, chloroquine, suppressed CpG DNA-induced MMP-9 expression and its activity. Moreover, CpG DNA induced the phosphorylation of ERK and the inhibition of ERK by U0126 suppressed CpG DNA-induced MMP-9 expression and its activity. CpG DNA stimulated $I{\kappa}B-{\alpha}$ degradation and luciferase activity. In addition, pretreatment of SN-50, the inhibitor of NF${\kappa}B$, strongly blocked the CpG DNA-induced MMP-9 expression and activity. Conclusion: These observations suggest that CpG DNA may play important roles in the activation of macrophages by regulating the production of MMP-9 via the sequential TLR9-ERK-NF${\kappa}B$ signaling pathway.

Amine Oxidase Activity of the Human Lysyl Oxidase-Like 3 (LOXL3) Protein

  • Kim Young-Ho
    • Biomedical Science Letters
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    • v.12 no.3
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    • pp.185-190
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    • 2006
  • Lysyl oxidase (LOX) catalyzes the lysine-derived cross-links of fibrillar collagens and elastin in the extracellular matrix. Recent molecular cloning has revealed existence of a LOX family consisting of LOX and four lysyl oxidase-like proteins (LOXL, LOXL2, LOXL3 and LOXL4). Pathological conditions associated with impaired LOX activity in several heritable and acquired disorders lead to severe structural and functional abnormalities of cardiovascular tissues, such as occlusion of coronary arteries and aneurysms, suggesting an essential role for the LOX family proteins in the maintenance of the cardiovascular system. However, the specific roles of the lysyl oxidase-like proteins in normal and pathological conditions of the cardiovascular tissues have not been established yet. Here, I report that LOXL3, a novel member of the LOX family, is predominantly expressed in the aorta, with an amine oxidase activity toward collagen and elastin, suggesting an essential role of LOXL3 in the development and maintenance of the aorta.

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