• Title/Summary/Keyword: peroxynitrite ($ONOO^-$)

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Anti-Diabetic and Anti-Inflammatory Effects of Green and Red Kohlrabi Cultivars (Brassica oleracea var. gongylodes)

  • Jung, Hyun Ah;Karki, Subash;Ehom, Na-Yeon;Yoon, Mi-Hee;Kim, Eon Ji;Choi, Jae Sue
    • Preventive Nutrition and Food Science
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    • v.19 no.4
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    • pp.281-290
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    • 2014
  • The aim of the present study was to evaluate the anti-diabetic, anti-inflammatory, antioxidant potential, and total phenolic content (TPC) of green and red kohlrabi cultivars. Anti-diabetic and anti-inflammatory activities were evaluated via protein tyrosine phosphatase (PTP1B) and rat lens aldose reductase inhibitory assays and cell-based lipopolysaccharide (LPS)-induced nitric oxide (NO) inhibitory assays in RAW 264.7 murine macrophages. In addition, scavenging assays using 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical, 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt (ABTS) radical, and peroxynitrite ($ONOO^-$) were used to evaluate antioxidant potential and TPC was selected to assess phytochemical characteristics. Between the two kohlrabi cultivars, red kohlrabi (RK) had two times more TPC than green kohlrabi (GK) and showed significant antioxidant effects in DPPH, ABTS, and $ONOO^-$ scavenging assays. Likewise, methanol (MeOH) extracts of RK and GK inhibited LPS-induced NO production in a dose dependent manner that was further clarified by suppression of iNOS and COX-2 protein production. The MeOH extracts of RK and GK exhibited potent inhibitory activities against PTP1B with the corresponding $IC_{50}$ values of $207{\pm}3.48$ and $287{\pm}3.22{\mu}g/mL$, respectively. Interestingly, the RK MeOH extract exhibited significantly stronger anti-inflammatory, anti-diabetic, and antioxidant effects than that of GK MeOH extract. As a result, our study establishes that RK extract with a higher TPC might be useful as a potent anti-diabetic, antioxidant, and anti-inflammatory agent.

Cytoprotective action of Rubi Fructus by modulation of Reactive Oxygen Species, peroxynitrite and $Ca^{2+}$ (복분자(覆盆子)의 세포내 ROS, $ONOO^-$ 생성 및 $Ca^{2+}$ 증가 억제에 의한 혈관내피세포 보호작용)

  • Lee, Cheol-Woong;Jeong, Ji-Cheon
    • The Journal of Internal Korean Medicine
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    • v.26 no.3
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    • pp.615-625
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    • 2005
  • Objectives : Poststroke depression is a frequent and specific entity that impaires the rehabilliation and functional recovery of patients with hemiplegia. The author evaluated the effect of Banhahubak-tang(Banxiahoupotang) in patients with poststroke depression. Methods : 38 patients suffering from poststroke depression(determined by Diagnostic and Statistical Manual of Mental Disorders, revised. 3rd edition. and Beck Depression Inventory[BDI] cutoff $point{\geqq}10$) in Kyunghee Oriental hospital were randomized into two groups; treatment group(n=19) and control group(n=19). The treatment group was prescribed with Banhahubak-tang(Banxiahoupotang) three times a day fur a week. Control troop was prescribed with other herbal medicines used for stroke Patients three times a day for a week. Patients were evaluated by use of BDI scale, Modified Barthel Index, Depression of Ki score, Yin syndrome score, and Yang syndrome score. Among 38 patients, 24 patients got BDI scores above 21, which is the cut-off score for depression in Korean. The same procedures and assessments described above were applied. Results : Treatment group did not significantly improve compared with control group. Results yielded only slight significance (P=0.086). Especially. patients with poststroke depression as yin syndrome improved more significantly on BDI than those classified as yang syndrome. When BDI cutoff point for depression was defined as being ${\geq}\;21$, treatment group did not significantly improve compared with control group(P=0.114). However, patients with poststroke depression classified as yin syndrome were also significantly improved on BDI than those classified as yang syndrome. Conclusions : This study suggests that Banhahubak-tang(Banxiahoupotang) is significantly effective in patients with poststroke depression classified as yin syndrome.

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Anti-inflammatory Activity of Medicinal Plant Extracts (약용식물자원 추출물의 항염증활성)

  • Lee, Seung-Eun;Lee, Jeong-Hoon;Kim, Jin-Kyung;Kim, Geum-Sook;Kim, Young-Ok;Soe, Jin-Sook;Choi, Je-Hun;Lee, Eun-Suk;Noh, Hyung-Jun;Kim, Seung-Yu
    • Korean Journal of Medicinal Crop Science
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    • v.19 no.4
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    • pp.217-226
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    • 2011
  • The study was conducted to investigate candidate materials as anti-inflammation agent from plant resources. Activities of 33 plant parts extracts with the final concentration of 5${\mu}g/ml$ were evaluated on the several inflammation-related markers such as the release of proinflammatoty cytokine [tumor necrosis factor-alpha (TNF-${\alpha}$) & interleukin-6 (IL-6)], nitric oxide (NO), the expression of inducible nitric oxide synthase (iNOS), cyclooxygenase-2 (COX-2) and inhibitor of nuclear factor kappa-B alpha ($I{\kappa}-B{\alpha}$) in lipopolysaccharide (LPS)-treated RAW 264.7 cells. The extracts in the final concentration of 10 ${\mu}g/ml$ were also screened on peroxynitrite (ONOO$^-$) scavenging activity. Eleven extracts selected from the screening assay were verified on the inhibition activity on peroxynitrite and total reactive species oxygen (ROS) in the several concentrations. As results, Alpinia officinarum Hance (rhizome), Inula britannica var. chinensis Regel (flower), Ulmus arvifolia Jacq (trunk peel) and Aster scaber Thunb. (aerial part) showed comparatively potent anti-inflammatory activities in vitro cells or chemical level systems, and then these four plant parts should be studied on the antiinflammatory mechanism by further studies.

Formation of Reactive Species Enhanced by H2O2 Addition in the Photodecomposition of N-Nitrosodimethylamine (NDMA)

  • Kwon, Bum Gun;Kim, Jong-Oh;Kwon, Joong-Keun
    • Environmental Engineering Research
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    • v.18 no.1
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    • pp.29-35
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    • 2013
  • This study noted that the actual mechanism of N-nitrosodimethylamine (NDMA) photodecomposition in the presence of $H_2O_2$ is missing from the previous works. This study investigated a key unknown reactive species (URS) enhanced by the addition of $H_2O_2$ during the photolysis of NDMA with $H_2O_2$, not hydroxyl radicals. In order to provide experimental evidences in support of URS formation, we have mainly used p-nitrosodimethylaniline, methanol, and benzoic acid as well-known probes of ${\cdot}OH$ in this study. Both loss of PNDA and formation of hydroxybenzoic acids were dependent on NDMA concentrations during the photolysis in a constant concentration of $H_2O_2$. In particular, competition kinetics showed that the relative reactivity of an URS was at least identical with ${\cdot}OH$-like reactivity. In addition, the decay of NDMA was estimated to be about 65% by the direct UV light and about 35% by the reactive species or URS generated through the photolysis of NDMA and $H_2O_2$. Therefore, our data suggest that a highly oxidizing URS is formed in the photolysis of NDMA with $H_2O_2$, which could be peroxynitrite ($ONOO^-$) as a potent oxidant by itself as well as a source of ${\cdot}OH$.

The Anticarcinogenic and Antioxidative Activity of Hemicentrotus pulacherrimus Fractions in Various Cancer Cells. (말똥성게 분획물에 의한 항 발암 및 항산화 효과)

  • Shin, Mi-Ok;Bae, Song-Ja
    • Journal of Life Science
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    • v.19 no.5
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    • pp.607-614
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    • 2009
  • This study was carried out to investigate the anticarcinogenic and antioxidative activities of Hemicentrotus pulacherrimus (HP). HP was extracted with methanol (HPM), which was then further fractionated into four sub-fractions by using the solvent partition method, affording methanol (HPMM), hexane (HPMH), butanol (HPMB) and aqueous (HPMA) soluble fractions. We determined the anticarcinogenic activities of these four fractions in four kinds of cancer cell lines, such as HepG2, HT29, MCF-7 and B16-F10, by MTT assay. Among various fractions from HPM, the HPMH showed the strongest growth inhibition effect. We also determined the inductive effect on quinone reductase (QR) of HP fractions. HPMB fraction exhibited strong inductive effects in HepG2 cells at a level of 90 ${\mu}g/ml$, showing inductive indexes of 2.26 compared to the control value of 1.0. The antioxidant activities of fractions from HP were also investigated by measuring the scavenging activities of HP against reactive oxygen speicies (ROS), peroxynitrite (ONOO-) and NO. Among the various solvent fractions, HPMH fractions displayed marked antioxidative activities.

Effects of Prunellae Herba on the H2O2-Treated LLC-PK Cell's Redox Status and NF-κB Signaling (H2O2 처리된 LLC-PK₁세포에서 Redox Status 및 NF-κB Signaling에 대한 하고초(夏枯草)의 효과)

  • Son, Jong Seok;Jeong, Ji Cheon
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.30 no.4
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    • pp.242-249
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    • 2016
  • This study was to investigate the anti-inflammatory effects of Prunellae Herba(PH). The generation of superoxide anion radical (․O2-), nitric oxide (NO), peroxynitrite (ONOO-) and Prostaglandin E₂(PGE2) were measured in the H2O2-Treated renal epithelial cells(LLC-PK1 cell) of mouse. And the effects of Prunellae Spica on the expression of NF-κB (p50, p65), IKK-α, phospho-IκB-α and inflammation-related proteins, COX-2, iNOS, IL-1β and VCAM-1, were examined by western blot. The fluorescent probes, 2',7'-dichlorodihydrofluorescein diacetate (DCFDA), 4,5-diaminofluorescein (DAF-2) and dihyldrorhodamine 123 (DHR 123) were used to estimate the scavenging effect of Prunellae Spica on ․O2-, NO, ONOO-. Western blot was conducted to assess the protein expression levels of NF-κB (p50, p65), IKK-α, phospho-IκB-α, inflammation-related proteins, COX-2, iNOS, IL-1β, VCAM-1. PH inhibited H2O2-treated cell death dose-dependently. It reduced the generation of ·O2-, NO, ONOO- and PGE₂ in the H2O2-treated renal epitheial cells(LLC-PK1 cell) of mouse in vitro. PH reduced the expression of NF-κB, IKK-α, phospho-IκB-α, COX-2, iNOS, IL-1β and VCAM-1 genes through means of decreasing activation of NF-κB signaling as well. According to these results, PH has an antioxidative activity and anti-inflammatory effect by regulating the NF-κB pathway. This suggest that PH is expected to be used to regulating inflammatory process and treating inflammation-related disease.

The Effect of Patriniae Radix on the Oxidative Stress and the NF-${\kappa}B$ Signaling in Mouse LLC-$PK_1$ Cell (Mouse의 신장상피세포에서 패장(敗醬)추출물이 산화 스트레스 및 NF-${\kappa}B$ signaling에 미치는 영향)

  • Kim, Hyun-Young;Jang, Soo-Young;Choi, Gyu-Ho;Shin, Hyeon-Cheol
    • The Journal of Internal Korean Medicine
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    • v.31 no.1
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    • pp.153-165
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    • 2010
  • Objectives : The aims of this study were to investigate the cytoprotective, antioxidative and inflammation genes inhibitory effects of Patriniae Radix on the mouse LLC-$PK_1$ cells (renal epithelial cells). Methods : The cytoprotective effect of Patriniae Radix was evaluated by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. The antioxidative effect was measured in terms of generation amount of superoxide anion radical (${\cdot}{O_2}^-$) by 2',7'-dichlorodihydrofluorescein diacetate (DCFDA), nitric oxide (NO) by 4,5-diaminofluorescein (DAF-2), peroxynitrite ($ONOO^-$) by dihyldrorhodamine 123 (DHR 123) and prostaglandin $E_2$ ($PGE_2$) by $PGE_2$ immunoassay on $H_2O_2$-treated LLC-$PK_1$ cells. For measuring of inflammation genes inhibitory effects, western blot was performed to detect IKK-$\alpha$, phospho-$I{\kappa}B-\alpha$, NF-${\kappa}B$ (p50, p65), COX-2, iNOS, IL-$1{\beta}$ and VCAM-1 protein level in cytosol fractions from LLC-$PK_1$ cells. Results : Patriniae Radix extract reduced the $H_2O_2$-induced cell death and inhibited the amount of $H_2O_2$-induced ${\cdot}{O_2}^-$, NO, $ONOO^-$, $PGE_2$ generation dose-dependently on the mouse LLC-$PK_1$ cells in vitro. Also Patriniae Radix extract inhibited the expression of IKK-$\alpha$, phospho-$I{\kappa}B-\alpha$, COX-2, iNOS, IL-$1\beta$ and VCAM-1 genes dose-dependently by means of decreasing activation of NF-${\kappa}B$. Conclusions : According to above results, it was identified that Patriniae Radix had the cytoprotective, antioxidative and inflammation genes inhibitory effects. So it was suggested that Patriniae Radix would be effective to the treatment for the inflammatory process and inflammation-related diseases.

Screening of Natural Resources with Inhibitory Activity on Free Radicals and Advanced Glycation end Products (AGEs) Formation (천연자원의 라디칼 소거능과 최종당화산물의 생성저해활성 검색)

  • Kim, Min-Suk;Kim, Dong-Wook;Rhyu, Dong-Young
    • Korean Journal of Pharmacognosy
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    • v.37 no.4 s.147
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    • pp.307-313
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    • 2006
  • Reactive oxygen species (ROS) and formation of advanced glycation end products (AGEs) play an important role in the pathogenesis of diabetic nephropathy by hyperglycemia. To find natural agents improving diabetic nephropathy, 63 natural resources which used to the treatment of diabetes mellitus in a folk remedy were investigated with an in vitro system employing radical scavenging activity and inhibitory activity of AGEs formation. In results, the extracts of Aspalathus linearis, Rubus coreanus, Rosa rugosa, and Epimedium koreanum significantly inhibited the 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical with $IC_{50}$ values less than $10{\mu}g/ml$. The extracts of Zea mays, Cucurbita moschata, Cudrania tricuspidata, and Aspalathus linearis effectively reduced the formation of AGEs compared with the positive control $N-acetyl-_L-cystenine$ (NAC) and aminoguanidine (AG). In addition, the extracts of Aspalathus linearis, Commelina communis, Cornus officinalis, and Lespodeza cuneata showed the all inhibitory activity against DPPH radical and AGEs formation. Also, these resources definitely showed the radical scavenging activity against peroxynitrite $(ONOO^-)$ and hydroxyl radical $({\cdot}OH)$ relating to high glucose-induced ROS production. Thus, these results suggest that some natural resources may regulate the initiation and progression of diabetic nephropathy through inhibition of ROS production and AGEs formation.

Flavonoids Differentially Modulate Nitric Oxide Production Pathways in Lipopolysaccharide-Activated RAW264.7 Cells

  • Kim Ae Ra;Cho Jae Youl;Zou Yani;Choi Jae Sue;Chung Hae Young
    • Archives of Pharmacal Research
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    • v.28 no.3
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    • pp.297-304
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    • 2005
  • Naturally occurring flavonoids are known to modulate various inflammatory and immune processes. Based on structural property, in this study, molecular mechanism of flavonoids in modulating nitric oxide (NO) production and its signaling pathway were investigated using lipopolysaccharide (LPS)-activated RAW264.7 cells. Although flavonol-typed flavonoids (kaempferol and quercetin) more potently scavenged reactivity of nitric oxide ($\cdot$NO) as well as peroxynitrite (ONOO$\kappa$) than isoflavones (genistein and genistin), kaempferol, quercetin and genistein showed a little difference in inhibition of both inducible NO synthase expression and NO production, with IC$_{50}$ values of 13.9, 20.1 and 26.8 $\mu$M. However, there was a striking pattern related to structural feature in modulation of LPS-mediated signaling pathways. Thus, flavonols only inhibited transcription factor AP-1 activation, whereas isoflavones suppressed the DNA binding activation of NF-$\kappa$B and C/EBP$\beta$. Therefore, these data suggest that structural feature may be linked to decide drugs target molecule in LPS-mediated signaling pathways, rather than its potency.

Antioxidant Activity of Extract from Walnut Uuglans sinensis Dode) and Its Protective Effect on Cell Injury and Lipid Peroxidation in Renal Cortical Slices (호두 추출물의 항산화 활성과 신피질에서 세포 손상과 지질과산화 방지효과)

  • Bae Kae Sun;Hwang Eul Chul;Kwon Chae Hwa;Kim Soon Hee;Choi Chun Whan
    • Journal of Life Science
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    • v.15 no.1 s.68
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    • pp.106-111
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    • 2005
  • To investigate the antioxidant activity of extract from the raw walnut, Juglans sinensis Dode, we prepared five fractions (methanol (MeOH), dichloromethane $(CH_2Cl_2)$, ethyl acetate (EtOAc), n-buthanol (n-BuOH) and dehydrogen monooxide $(H_2O)$ fractions) and examined. The effect of walnut extract on the oxidative stress was investigated in vitro. The DPPH (2,2-Di (4-tert-octylphenyl)-1-picrylhydrazyl) free radical scavenging activity of extract from raw walnut was shown in the following order: $EtOAc\;fraction layer. The result showed that the highest activity $(0.56{\mu}g/ml,\;IC_{50}.)$ was observed in EtOAc fraction, whereas n-BuOH fraction, MeOH fraction, $CH_2O_2$ fraction and $H_2O$ layer of $IC_{50}$ were $2.34{\mu}g//ml,\;3.88{\mu}g/ml,\;8.06{\mu}g/ml,\;and\;8.19{\mu}g/ml$, respectively. The radical scavenging activity assay of each fraction showed that the antioxidative activity was observed in the following order: EtOAc fraction $(74.27\pm1.56\%)>MeOH\;fraction\;(60.76\pm3.4\%)>n-BuOH\;fraction\;(59.32\pm0.88\%)>H_2O\;layer\;(41.69\pm2.06\%)$. These results revealed that all fractions, except for $CH_2Cl_2$ fraction, showed high antioxidative activity. Furthermore, the peroxynitrite $(ONOO^-)$ scavenging activity was assayed in each fraction. The result showed that the $ONOO^-$ scavenging activity of EtOAc fraction, MeOH fraction and n-BuOH fraction from raw walnut was $95.14\pm0.36\%,\; 90.02\pm1.19\%\;and\;89.41\pm0.81\%$, respectively. The tert-butylhydroperoxide (t-BHP) treatment in vitro increased lactate dehydrogenase release and lipid peroxidation in renal cortical slices. Such changes were completely prevented by addition of MeOH fraction, EtOAc fraction and n-BuOH fraction of walnut. These results indicate that the walnut extract exerts the benedicial effect against t-BHP-induced cell injury and its effect may be due to antioxidant action. In addition, it is suggested that walnut extract might be developed as the effective scavenger for the prevention of oxidative stress.