• Title/Summary/Keyword: superoxide generation

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Protective effects of Carthamus tinctorius L. seed on C6 glial cells treated with ethanol (홍화씨 추출물의 in vitro 항산화 및 ethanol로 손상을 유도한 C6 신경교세포 보호 효과)

  • Choi, Seung Hak;Park, Chan Hum;Cho, Eun Ju;Kim, Ji Hyun;Seo, Weon Taek
    • Journal of Applied Biological Chemistry
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    • v.64 no.1
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    • pp.69-74
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    • 2021
  • Chronic alcohol is responsible for oxidative stress and neurodegenerative diseases such as dementia. In the present study, we investigated the antioxidant activity and protective effects of seed of Carthamus tinctorius L. on ethanol-induced C6 glial cells. Antioxidant effect of seed of C. tinctorius L. was measured by scavenging activity of 1,1-diphenyl-2-prcrylhydrazy (DPPH), hydroxyl radical (·OH), superoxide radical, and nitric oxide. The seed of C. tinctorius L. extract showed significant radical scavenging activities in a concentration-dependent manner. In particular, it revealed strong DPPH and ·OH scavenging activity, displaying more than 80% at 500 and 100 ㎍/mL, respectively. Treatment of 500 mM ethanol to C6 glial cell led to decline of cell viability and elevation of reactive oxygen species (ROS) generation. However, seed of C. tinctorius L.-treated groups significantly increased cell viability and decreased ROS levels, compared to ethanol-induced control group. These results suggest that seed of C. tinctorius L. would have protective effect against neuronal oxidative stress induced by alcohol.

Long-term recombinant interferon-γ treatment in 2 cases of osteopetrosis (장기간 인터페론 감마로 치료한 골화석증 2례)

  • Kim, Dong-Yun;Han, Dong-Kyun;Baek, Hee-Jo;Jung, Sung-Taek;Kook, Hoon;Hwang, Tai-Ju
    • Clinical and Experimental Pediatrics
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    • v.50 no.11
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    • pp.1129-1133
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    • 2007
  • Osteopetrosis, a rare osteosclerotic bone disease characterized by a defect in osteoclast function and the reduced generation of superoxide by leukocytes, can be classified into several types based on their mode of inheritance, age of onset, severity, and associated clinical symptoms. Stem cell transplantation is the only curative therapy for the infantile malignant type, although alternative treatments, such as corticosteroids, calcitriol, and interferon (IFN)-${\gamma}$ have been attempted in patients with milder clinical types. In addition, IFN-${\gamma}$ therapy has been reported to increase bone resorption and hematopoiesis and to improve leukocyte function. Here, we present the cases of two patients with osteopetrosis who benefited from either 3 or 6 years of INF-${\gamma}$ therapy that resulted in improved blood counts and no further pathological fractures.

Experimental Studies on the Effect of Gamibaegi-eum

  • Kim Won-Ill
    • The Journal of Korean Medicine
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    • v.25 no.4
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    • pp.61-78
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    • 2004
  • Objective : This study was undertaken to determine whether Gamibaegi-eum (BGU) in vitro and in vivo exerts a beneficial effect against cell injury induced by reactive oxygen species (ROS) in the human intestine. Methods : Effects of BGU in vitro on cell injury were examined using Caco-2 cells, cultured human intestinal cell line. Exposure of cells to H₂O₂ induced increases in the loss of cell viability in a time and dose-dependent fashion. Results : BGU prevented H₂O₂-induced cell death and its effect was dose-dependent over a concentration range of 0.05­1%. H₂O₂-induced cell death was prevented by catalase, the hydrogen peroxide scavenger enzyme, and deferoxamine, the iron chelator. However, the potent antioxidant DPPD did not affect H₂O₂-induced cell death. H₂O₂ increased lipid peroxidation, which was inhibited by BGU and DPPD. H₂O₂ caused DNA damage in a dose-dependent manner, which was prevented by BGU, catalase, and deferoxamine, but not DPPD. BGU restored ATP depletion induced by H₂O₂. BGU inhibited generation of superoxide and H₂O₂ and scavenged directly H₂O₂. Oral administration of mepirizole in vivo at a dose of 200mg/kg resulted in ulcer lesions in the stomach and the proximal duodenum. Pretreatment of BGU(0.1%/kg, orally) and catalase (800Units/kg, i.v.) significantly decreased the size of ulcers. Mepirizole increased lipid peroxidation in the mucosa of the duodenum, suggesting an involvement of ROS. Pretreatment of BGU and catalase significantly inhibited lipid peroxidation induced by mepirizole. Morphological studies showed that mepirizole treatment causes duodenal injury and its effect is prevented by BGU. Conclusion : These results indicate that BGU exerts a protective effect against cell injury in vitro and in vivo through antioxidant action. The present study suggests that BGU may playa therapeutic role in the treatment of human gastrointestinal diseases mediated by ROS.

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Anti-aging Effect on Skin with the needles of red pine, Pinus densiflora (적송엽(赤松葉)이 피부(皮膚)의 노화(老化)에 미치는 영향(影響))

  • Park, Seong-Kyu;Lee, Jong-Chan;Ahn, Soo-Mi;Lee, Jin-Young;Kim, Youn-Joon;Hwang, Jae-Sung;Lee, Byeong-Gon;Chang, Ih-Seoup
    • The Korea Journal of Herbology
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    • v.20 no.4
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    • pp.1-10
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    • 2005
  • Objectives : We investigated the anti-aging effect on skin with the extract of the needles of red pine, Pinus densiflora. Methods : We measured various effects related to skin such as the anti-oxidant effect, the protection against ultraviolet (UV) irradiation, the inhibition of reactive oxygen species (ROS) generation, the induction of heat shock protein 70 (HSP70), the reduction of matrix metalloproteinase-2 (MMP-2) synthesis and senescent cell. Results : The results were as follows : The extract of the needles of red pine (RP) had the potent anti-oxidant effect and the ROS scavenging effect. Also RP preserved the systemic anti-oxidant enzyme system (superoxide dismutase and catalase) from UVB irradiation. RP protected the cell membrane from the damages induced by UVB irradiation. RP induced HSP70, a mediator of resistance to UVB irradiation. RP reduced the synthesis of MMP-2 induced by UVB irradiation. And RP inhibited the amount of senescent-associated (SA) ${\beta}-galactosidase$ staining, as a marker of replicative senescence. Conclusions : The results of our study indicate that the extract of the needles of red pine, Pinus densiflora, has anti-aging effects on skin.

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Role of tetrahydrobiopterin in dopaminergic cell death: Relevance to Parkinson's disease

  • Choi, Hyun-Jin;Hwang, On-You
    • Proceedings of the Korean Society of Applied Pharmacology
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    • 2005.04a
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    • pp.53-60
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    • 2005
  • Parkinson's disease (PD) is a progressive neurodegenerative disorder affecting $1\%$ of the population above the age of 65 and is characterized by a selective loss of dopaminergic neurons in the substantia nigra pars compacta. Although the underlying cause of dopaminergic cell death or the mechanism by which these cells degenerate is still not clearly understood, oxidative stress, mitochondrial dysfunction, and protein misfolding are thought to play important roles in the dopaminergic degeneration in PD. Tetrahydrobiopterin (BH4) is synthesized exclusively in the monoaminergic, including dopaminergic, cells and serves as an endogenous and obligatory cofactor for syntheses of the potential oxidative stressors dopamine and nitric oxide. In addition to its contribution toward the syntheses of these two potentially toxic molecules, BH4 itself can directly generate oxidative stress. BH4 undergoes oxidation during the hydroxylation reaction as well as nonenzymatic autooxidation to produce hydrogen peroxide and superoxide radical. We have previously suggested BH4 as an endogenous molecule responsible for the dopaminergic neurodegeneration. BH4 exerts selective toxicity to dopamine-producing cells via generation of oxidative stress, mitochondrial dysfunction, and apoptosis. BH4 also induces morphological, biochemical, and behavioral characteristics associated with PD in vivo. BH4 as well as enzyme activity and gene expression of GTP cyclohydrolase I, the rate-limiting enzyme in BH4 synthesis pathway, are readily upregulated by cellular changes such as calcium influx and by various stimuli including stress situations. This points to the possibility that cellular availability of BH4 might be increased in aberrant conditions, leading to increased extracellular BH4 subsequent degeneration. The fact that BH4 is specifically and endogenously synthesized in dopaminergic cells, Is readily upregulated, and generates oxidative stress-related cell death provides physical relevance of this molecule as an attractive candidate with which to explain the mechanism of pathogenesis of PD.

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Anti-oxidant and Anti-inflammation Activities of Prunus persica Flos (도화(桃花, Prunus persica Flos)의 항산화 및 항염증 활성)

  • Lee, Jin-Young;An, Bong-Jeun
    • Journal of Applied Biological Chemistry
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    • v.53 no.3
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    • pp.162-169
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    • 2010
  • The solvent extracts of Prunus persica Flos were investigated for the activities of anti-oxidant and anti-inflammation to apply as a functional ingredient for cosmetic products. The electron donating ability of both ethanol (PPE) or acetone (PPA) extracts of P. persica Flos was above 90.0% at the concentration of 500ppm. The superoxide dismutase (SOD)-like activity of P. persica Flos extracts (PPE, PPA) were approximately 40.0% at 1,000 ppm. The xanthine oxidase inhibitory effect of P. persica Flos extracts (PPE, PPA) was approximately 30.0% at 1,000 ppm and equivalent to that of ascorbic acid. Hyaluronidase inhibition activity related to the anti-inflammation effect was 35.0% with the treatment of P. persica Flos extracts (PPW, PPE, PPA) at 1,000 ppm, respectively. In the experiment of anti-inflammation effect, P. persica Flos extracts (PPW, PPE, PPA) inhibited the generation of nitric oxide. In the antimicrobial activity test against the human skin-resident microflora such as Staphylococcus epidermidis and Propionibacterium acnes, a clear zone was identified from 4mg/disc in P. persica Flos (PPE) extract.

Antioxidant activities of Erythrina stricta Roxb.using various in vitro and ex vivo models

  • AsokKumar, K;UmaMaheswari, M;Sivashanmugam, AT;SubhadraDevi, V;Subhashini, N;Ravi, TK
    • Advances in Traditional Medicine
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    • v.8 no.3
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    • pp.266-278
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    • 2008
  • Erythrina stricta, a deciduous tree widely used traditionally in indigenous system of medicine for various ailments such as rheumatism, fever, leprosy, epilepsy etc. The leaves of Erythrina stricta was extracted with ethanol (70%) and used for the evaluation of various in vitro antioxidant assays which includes H - donor activity, nitric oxide scavenging, superoxide anion scavenging, reducing ability, hydroxyl radical, hydrogen peroxide scavenging, total phenolic content, total flavonoid content, total antioxidant activity by thiocyanate and phosphomolybdenum method, metal chelating, $\beta$-carotene bleaching, total peroxy radical assays. The pro-oxidant activity was measured using bleomycin-dependent DNA damage. Ex vivo models like lipid peroxidation and erythrocyte haemolysis were also used to study the antioxidant property of the extract. The various antioxidant activities were compared with suitable standard antioxidants such as ascorbic acid, butylated hydroxyl toluene, $\alpha$-tocopherol, curcumin, quercetin and Trolox. The generation of free radicals viz. $O_2^{{\cdot}-}$, $OH^{\cdot}$, $H_2O_2$, $NO^{\cdot}$ and peroxyl radicals were effectively scavenged by the ethanolic extract of Erythrina stricta. In all the methods, the extract offered strong antioxidant activity in a concentration dependent manner. The total phenolic content, flavonoid content and total antioxidant activity in Erythrina stricta were determined as microgram (g) pyrocatechol, quercetin and $\alpha$-tocopherol equivalent/mg respectively. The extract did not exhibit any prooxidant activity when compared with ascorbic acid. The results obtained in the present study clearly indicates that Erythrina stricta scavenges free radicals and reduces lipid peroxidation, ameliorating the damage imposed by oxidative stress in different disease conditions and serve as a potential source of natural antioxidant.

Effects of Ojung-hwan on Blood Glucose, Hyperlipidemia, Polyol Pathway and Antioxidative Mechanism in Ob/ob Mouse (Ob/ob mouse에서 오정환(五精丸)이 혈당, 고지혈증, Polyol Pathway 및 항산화작용에 미치는 영향)

  • Gong, Tae-Hyun;Jeong, Ji-Cheon
    • The Journal of Korean Medicine
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    • v.28 no.3 s.71
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    • pp.57-69
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    • 2007
  • Objectives : Diabetes is a disease in which the body does not produce or properly use insulin. Etiological studies of diabetes and its complications have shown that oxidative stress might play a major role. Therefore, many methods have been tried to regulate free oxygen radicals for treating diabetes and its complications. Ojung-hwan, composed of five crude herbs, has been considered effective for treating symptoms of aging. In male ob/ob mouse of severe obesity, hyperinsulinemia and hyperlipidemia, which are features of NIDDM, the hyperglycemic activities and mechanisms of Ojung-hwan were examined. Methods : Mice were grouped and treated for 5 weeks as follows. Both the lean (C57/BL6J black mice) and diabetic (ob/ob mice) control groups received standard chow. The experimental groups were fed a diet of chow supplemented with 30 and 90 mg Ojung-hwan per 1 kg of body weight for 14 days. The effects of Ojung-hwan extract on the ob/ob mice were observed by measuring the serum levels of glucose, insulin, lipid components, and the kidney levels of superoxide anion radical (${\cdot}\;O{_2}{^-}$), MDA+HAE, GSH/GSSG ratio, and also the enzyme activities involved in polyol pathway. Results : Ojung-hwan lowered the levels of serum glucose and insulin in a dose-dependent manner. Total cholesterol, triglyceride and free fatty acid levels decreased, while the HDL-cholesterol level increased, in Ojung-hwan treated groups. Renal aldose reductase and sorbitol dehydrogenase activities increased in the ob/ob mice, whereas they were inhibited in the Ojung-hwan treated groups. Ojung-hwan inhibited the generation of ${\cdot}\;O{_2}{^-}$ in the kidney. Finally, MDA+HAE levels increased and GSH/GSSG ratio decreased in the ob/ob mice, whereas they improved in the Ojung-hwan treated groups. Conclusions : Ojung-hwan showed antidiabetic and antihyperlipidemic activities by regulating theactivities of polyol pathway enzymes, scavenging reactive oxygen species and reducing the MDA+HAE levels in the ob/ob mice.

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Activated Phenoloxidase Interacts with A Novel Glycine-rich Protein on the Yeast Two-hybrid System

  • Lee, Sun-Woo;Lee, Hyun-Seong;Kim, Eun-Jun;Yoo, Mi-Ae;Lee, Bok-Luel
    • BMB Reports
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    • v.34 no.1
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    • pp.15-20
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    • 2001
  • One of the innate immune reactions in invertebrates is the pro-phenoloxidase (pro-PO) activation system that is involved in the generation of superoxide, melanin synthesis, and the subsequent sequestration of foreign matter entering the hemocoel of the invertebrates. However, the molecular mechanism of this biological reaction is still obscure. To expand our understanding of the biological roles of the pro-PO activation system in invertebrates, we performed a yeast two-hybrid screening by using three regions of pro-PO as bait and a yeast two-hybrid cDNA library from Tenebrio molitor larvae as prey We isolated a novel partial cDNA clone that encodes a glycine-rich protein that interacted with the active phenoloxidase (termed phenoloxidase interacting protein, POIP). POIP consists of two domains: One is an N-terminal unique domain and the other is a C-terminal glycine-rich domain. The C-terminal glycine-rich domain showed sequential homology with those of insect antifungal proteins. Also, the yeast two-hybrid screen in a reverse orientation (using POIP as bait) yielded PO, suggesting that the PO-POIP interaction is specific. By using a 315 bP PCR fragment of the N-terminal unique region of POIP, we cloned the full-length cDNA of POIP from the Tenebruo cDNA library constructed by using E. coli injected larvae. The interaction analysis between PO, and a truncated fragment lacking the N-terminal unique region of POIP, indicated that the N-terminal unique region is necessary for interaction between PO and POIP. The expression level of the POIP mRNA is increased by bacterial injection into T. molitor larvae. This suggests that POIP might be engaged in the humoral defense reaction.

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Pretreatment with Lycopene Attenuates Oxidative Stress-Induced Apoptosis in Human Mesenchymal Stem Cells

  • Kim, Ji Yong;Lee, Jai-Sung;Han, Yong-Seok;Lee, Jun Hee;Bae, Inhyu;Yoon, Yeo Min;Kwon, Sang Mo;Lee, Sang Hun
    • Biomolecules & Therapeutics
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    • v.23 no.6
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    • pp.517-524
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    • 2015
  • Human mesenchymal stem cells (MSCs) have been used in cell-based therapy to promote revascularization after peripheral or myocardial ischemia. High levels of reactive oxygen species (ROS) are involved in the senescence and apoptosis of MSCs, causing defective neovascularization. Here, we examined the effect of the natural antioxidant lycopene on oxidative stress-induced apoptosis in MSCs. Although $H_2O_2$ ($200{\mu}M$) increased intracellular ROS levels in human MSCs, lycopene ($10{\mu}M$) pretreatment suppressed $H_2O_2$-induced ROS generation and increased survival. $H_2O_2$-induced ROS increased the levels of phosphorylated p38 mitogen activated protein kinase (MAPK), Jun-N-terminal kinase (JNK), ataxia telangiectasia mutated (ATM), and p53, which were inhibited by lycopene pretreatment. Furthermore, lycopene pretreatment decreased the expression of cleaved poly (ADP ribose) polymerase-1 (PARP-1) and caspase-3 and increased the expression of B-cell lymphoma 2 (Bcl-2) and Bcl-2-associated X protein (Bax), which were induced by $H_2O_2$ treatment. Moreover, lycopene significantly increased manganese superoxide dismutase (MnSOD) expression and decreased cellular ROS levels via the PI3K-Akt pathway. Our findings show that lycopene pretreatment prevents ischemic injury by suppressing apoptosis-associated signal pathway and enhancing anti-oxidant protein, suggesting that lycopene could be developed as a beneficial broad-spectrum agent for the successful MSC transplantation in ischemic diseases.