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1.
Cerebellar granule neurons (CGN) cultured in a medium containing 25 mM KCl and treated with staurosporine (ST) or transferred to a medium with 5 mM KCl (K5) die apoptotically. CGN death is mediated by an increase in reactive oxygen species (ROS) production. When CGN are treated with antioxidants all apoptotic parameters and cell death are markedly diminished, showing a central role for ROS in this process. Recently, it has been suggested that a possible ROS source involved in cell death is a NADPH oxidase. In that regard, we found expression in CGN of the components of NADPH proteins, p40phox, p47phox and p67phox, and p22phox, as well as three homologues of the catalytic subunit of this complex, NOX1, 2, and 4. The inhibition of NADPH oxidase with diphenylene iodonium or 4-(2-aminoethyl)benzenesulfonyl fluoride significantly reduced ROS production, NADPH oxidase activity, all the apoptotic events, and cell death induced by both K5 and ST. We conclude that ROS could be an early signal of apoptotic neuronal death and that NADPH oxidase, including NOX1, 2, and/or 4, could have a central role in apoptotic death induced by different conditions in these neurons. 相似文献
2.
Dan Gao Shanwei Nong Xiuqing Huang Yonggang Lu Hongye Zhao Yajun Lin Yong Man Shu Wang Jiefu Yang Jian Li 《The Journal of biological chemistry》2010,285(39):29965-29973
Elevated plasma free fatty acid (FFA) levels in obesity may play a pathogenic role in the development of insulin resistance. However, molecular mechanisms linking FFA to insulin resistance remain poorly understood. Oxidative stress acts as a link between FFA and hepatic insulin resistance. NADPH oxidase 3 (NOX3)-derived reactive oxygen species (ROS) may mediate the effect of TNF-α on hepatocytes, in particular the drop in cellular glycogen content. In the present study, we define the critical role of NOX3-derived ROS in insulin resistance in db/db mice and HepG2 cells treated with palmitate. The db/db mice displayed increased serum FFA levels, excess generation of ROS, and up-regulation of NOX3 expression, accompanied by increased lipid accumulation and impaired glycogen content in the liver. Similar results were obtained from palmitate-treated HepG2 cells. The exposure of palmitate elevated ROS production and NOX3 expression and, in turn, increased gluconeogenesis and reduced glycogen content in HepG2 cells. We found that palmitate induced hepatic insulin resistance through JNK and p38MAPK pathways, which are rescued by siRNA-mediated NOX3 reduction. In conclusion, our data demonstrate a critical role of NOX3-derived ROS in palmitate-induced insulin resistance in hepatocytes, indicating that NOX3 is the predominant source of palmitate-induced ROS generation and that NOX3-derived ROS may drive palmitate-induced hepatic insulin resistance through JNK and p38MAPK pathways. 相似文献
3.
Moretto MB Rossato JI Nogueira CW Zeni G Rocha JB 《Journal of biochemical and molecular toxicology》2003,17(3):154-160
By mediating the Ca(2+) influx, Ca(2+) channels play a central role in neurotransmission. Chemical agents that potentially interfere with Ca(2+) homeostasis are potential toxic agents. In the present investigation, changes in Ca(2+) influx into synaptosomes by organic forms of selenium and tellurium were examined under nondepolarizing and depolarizing conditions induced by high KCl concentration (135 mM) or by 4-aminopyridine (4-AP). Under nondepolarizing conditions, ebselen (400 micro M) increased Ca(2+) influx; diphenyl ditelluride (40-400 micro M) decreased Ca(2+) in all concentrations tested; and diphenyl diselenide decreased Ca(2+) influx at 40 and 100 micro M, but had no effect at 400 micro M. In the presence of KCl as depolarizing agent, ebselen and diphenyl ditelluride decreased Ca(2+) influx in a linear fashion. In contrast, diphenyl diselenide did not modify Ca(2+) influx into isolated nerve terminals. In the presence of 4-AP (3 mM) as depolarizing agent, ebselen (400 micro M) caused a significant increase, whereas diphenyl diselenide and diphenyl ditelluride inhibited Ca(2+) influx into synaptosomes. The results can be explained by the fact that the mechanism through which 4-AP and high K(+) induced elevation of intracellular Ca(2+) is not exactly coincident. The mechanism by which diphenyl ditelluride and ebselen interact with Ca(2+) channel is unknown, but may be related to reactivity with critical sulfhydryl groups in the protein complex. The results of the present study indicate that the effects of organochalcogenides were rather complex depending on the condition and the depolarizing agent used. 相似文献
4.
《Free radical research》2013,47(5):600-610
Abstractγ-Glutamyltransferase (GGT) plays a significant role in antioxidant defence and participates in the metabolism of glutathione (GSH). The enzyme is up-regulated after acute oxidative stress and during pro-oxidant periods, but the underlying regulatory mechanisms are not well known. The present investigation studied whether the endogenous reactive oxygen species (ROS) level was a determinant for GGT expression. A substantial amount of ROS is produced through the NADPH oxidase (NOX) system and knockdown of p22phox, a sub-unit of NOX1-4, resulted not only in reduced ROS levels but also in reduced GGT expression in human endometrial carcinoma cells. Phorbol-12-myristate-13-acetate (PMA) is an activator of NOX and it was found that PMA treatment of human colon carcinoma cells both increased cellular ROS levels and subsequently up-regulated GGT expression. On the other hand, the NOX inhibitor apocynin reduced ROS levels as well as GGT expression. The GGT mRNA sub-type A was increased after PMA-induced NOX activation. These results demonstrate that ROS generated from NOX enzymes are a significant determinant for GGT expression and activity. 相似文献
5.
Reactive oxygen species (ROS) generated upon collagen stimulation act as second messengers to propagate various platelet-activating events. Among the ROS-generating enzymes, NADPH oxidase (NOX) plays a prominent role in platelet activation. Thus, NOX has been suggested as a novel target for anti-platelet drug development. Although kaempferol has been identified as a NOX inhibitor, the influence of kaempferol on the activation of platelets and the underlying mechanism have never been investigated. Here, we studied the effects of kaempferol on NOX activation, ROS-dependent signaling pathways, and functional responses in collagen-stimulated platelets. Superoxide anion generation stimulated by collagen was significantly inhibited by kaempferol in a concentration-dependent manner. More importantly, kaempferol directly bound p47phox, a major regulatory subunit of NOX, and significantly inhibited collagen-induced phosphorylation of p47phox and NOX activation. In accordance with the inhibition of NOX, ROS-dependent inactivation of SH2 domain-containing protein tyrosine phosphatase-2 (SHP-2) was potently protected by kaempferol. Subsequently, the specific tyrosine phosphorylation of key components (Syk, Vav1, Btk, and PLCγ2) of collagen receptor signaling pathways was suppressed by kaempferol. Kaempferol also attenuated downstream responses, including cytosolic calcium elevation, P-selectin surface exposure, and integrin-αIIbβ3 activation. Ultimately, kaempferol inhibited platelet aggregation and adhesion in response to collagen in vitro and prolonged in vivo thrombotic response in carotid arteries of mice. This study shows that kaempferol impairs collagen-induced platelet activation through inhibition of NOX-derived ROS production and subsequent oxidative inactivation of SHP-2. This effect suggests that kaempferol has therapeutic potential for the prevention and treatment of thrombovascular diseases. 相似文献
6.
Suchismita Mazumdar Thankamani Marar Shine Devarajan Jyoti Patki 《Biochemistry and Biophysics Reports》2021
Clinical evidence suggests that type 2 diabetes therapy can greatly benefit from the suppression of reactive oxygen species generation and the activation or restoration of cellular antioxidant mechanisms. In human, NADPH oxidase (NOX) is the main producer of reactive oxygen species (ROS) that supress the activity of endogenous antioxidant enzymes. In the present study, the antioxidant potential of Gedunin was studied. In silico findings reveal its strong binding affinity with NOX5 C terminal HSP90 binding site that disrupts NOX5 stability and its ability to generate ROS, leading to restoration antioxidant enzymes activities. It was found that Gedunin suppressed hyperglycaemia induced oxidative stress in an in vitro RBC model and markedly reversed glucose induced changes including haemoglobin glycosylation and lipid peroxidation. A significant restoration of activities of cellular antioxidant enzymes; superoxide dismutase, catalase and glutathione peroxidase in the presence of Gedunin revealed its ability to reduce oxidative stress. These results substantiated Gedunin as a bona fide inhibitor of human NOX5 and a ROS scavenging antioxidant with promising therapeutic attributes including its natural origin and inhibition of multiple diabetic targets. 相似文献
7.
《Free radical research》2013,47(9):1033-1039
AbstractThis study aimed to examine the roles of reactive oxygen species (ROS) in cisplatin treatment of human prostate cancer cells; hormone-sensitive LNCaP and hormone-refractory PC3 and DU145 cells. Intracellular levels of ROS and H2O2 were measured and visualized using specific fluorescent probes. NADPH oxidase (NOX) activity was detected by lucigenin chemiluminescence assay. Expression levels of NOX isoforms were determined by semi-quantitative RT-PCR. Cisplatin treatment increased the intracellular levels of ROS and H2O2 in three prostate cancer cell lines. The increase was transient and robust in hormone-sensitive LNCaP cells compared with hormone-refractory PC3 and DU145 cells. Consistent with these findings, the NOX activity induced by cisplatin was higher in LNCaP cells than in PC3 and DU145 cells. Expression pattern of NOX isoforms varied among three cell lines and the NOX activity was independent of NOX expression. Taken together, we have shown that cisplatin induces production of ROS and H2O2 via NOX activation in human prostate cancer cell lines, which is most prominent in hormone-sensitive LNCaP cells. 相似文献
8.
γ-Glutamyltransferase (GGT) plays a significant role in antioxidant defence and participates in the metabolism of glutathione (GSH). The enzyme is up-regulated after acute oxidative stress and during pro-oxidant periods, but the underlying regulatory mechanisms are not well known. The present investigation studied whether the endogenous reactive oxygen species (ROS) level was a determinant for GGT expression. A substantial amount of ROS is produced through the NADPH oxidase (NOX) system and knockdown of p22phox, a sub-unit of NOX1-4, resulted not only in reduced ROS levels but also in reduced GGT expression in human endometrial carcinoma cells. Phorbol-12-myristate-13-acetate (PMA) is an activator of NOX and it was found that PMA treatment of human colon carcinoma cells both increased cellular ROS levels and subsequently up-regulated GGT expression. On the other hand, the NOX inhibitor apocynin reduced ROS levels as well as GGT expression. The GGT mRNA sub-type A was increased after PMA-induced NOX activation. These results demonstrate that ROS generated from NOX enzymes are a significant determinant for GGT expression and activity. 相似文献
9.
Evren Yildiztugay Ceyda Ozfidan-Konakci Mustafa Kucukoduk 《Journal of plant research》2013,126(5):729-742
Sphaerophysa kotschyana is a Turkish endemic and endangered plant that grows near Salt Lake, in Konya, Turkey. However, little is known about the ability of this plant to generate/remove reactive oxygen species (ROS) or its adaptive biochemical responses to saline environments. After exposure of S. kotschyana to 0, 150, and 300 mM NaCl for 7 and 14 days, we investigated (1) the activities and isozyme compositions of the antioxidant enzymes superoxide dismutase (SOD), catalase (CAT), peroxidase (POX), ascorbate peroxidase (APX), and glutathione reductase (GR); (2) the oxidative stress parameters NADPH oxidase (NOX) activity, lipid peroxidation (MDA), total ascorbate (tAsA) content, and total glutathione content (tGlut); and (3) ROS levels for superoxide anion radical (O 2 ·? ), hydrogen peroxide (H2O2), hydroxyl radicals (OH·), and histochemical staining of O 2 ·? and H2O2. H2O2 content increased after 14 days of salt stress, which was consistent with the results from histochemical staining and NOX activity measurements. In contrast, oxidative stress induced by 150 mM NaCl was more efficiently prevented, as indicated by low malondialdehyde (MDA) levels and especially at 7 days, by increased levels of SOD, POX, APX, and GR. However, at 300 mM NaCl, decreased levels of protective enzymes such as SOD, CAT, POX, and GR, particularly with long-term stress (14 days), resulted in limited ROS scavenging activity and increased MDA levels. Moreover, at 300 mM NaCl, the high H2O2 content caused oxidative damage rather than inducing protective responses against H2O2. These results suggest that S. kotschyana is potentially tolerant to salt-induced damage only at low salt concentrations. 相似文献
10.
11.
Maarten A. Ligtenberg ?zcan ??nar Rikard Holmdahl Dimitrios Mougiakakos Rolf Kiessling 《PloS one》2015,10(6)
Reactive oxygen species (ROS) produced by the inducible NADPH oxidase type 2 (NOX2) complex are essential for clearing certain infectious organisms but may also have a role in regulating inflammation and immune response. For example, ROS is involved in myeloid derived suppressor cell (MDSC)- and regulatory T cell (Treg) mediated T- and NK-cell suppression. However, abundant ROS produced within the tumor microenvironment, or by the tumor itself may also yield oxidative stress, which can blunt anti-tumor immune responses as well as eventually leading to tumor toxicity. In this study we aimed to decipher the role of NOX2-derived ROS in a chemically (by methylcholanthrene (MCA)) induced sarcoma model. Superoxide production by NOX2 requires the p47phox (NCF1) subunit to organize the formation of the NOX2 complex on the cell membrane. Homozygous mutant mice (NCF1*/*) have a functional loss of their super oxide burst while heterozygous mice (NCF1*/+) retain this key function. Mice harboring either a homo- or a heterozygous mutation were injected intramuscularly with MCA to induce sarcoma formation. We found that NOX2 functionality does not determine tumor incidence in the tested MCA model. Comprehensive immune monitoring in tumor bearing mice showed that infiltrating immune cells experienced an increase in their oxidative state regardless of the NOX2 functionality. While MCA-induced sarcomas where characterized by a Treg and MDSC accumulation, no significant differences could be found between NCF1*/* and NCF1*/+ mice. Furthermore, infiltrating T cells showed an increase in effector-memory cell phenotype markers in both NCF1*/* and NCF1*/+ mice. Tumors established from both NCF1*/* and NCF1*/+ mice were tested for their in vitro proliferative capacity as well as their resistance to cisplatin and radiation therapy, with no differences being recorded. Overall our findings indicate that NOX2 activity does not play a key role in tumor development or immune cell infiltration in the chemically induced MCA sarcoma model. 相似文献
12.
Background
Cycling and chronic tumor hypoxia are involved in tumor development and growth. However, the impact of cycling hypoxia and its molecular mechanism on glioblastoma multiforme (GBM) progression remain unclear.Methodology
Glioblastoma cell lines, GBM8401 and U87, and their xenografts were exposed to cycling hypoxic stress in vitro and in vivo. Reactive oxygen species (ROS) production in glioblastoma cells and xenografts was assayed by in vitro ROS analysis and in vivo molecular imaging studies. NADPH oxidase subunit 4 (Nox4) RNAi-knockdown technology was utilized to study the role of Nox4 in cycling hypoxia-mediated ROS production and tumor progression. Furthermore, glioblastoma cells were stably transfected with a retroviral vector bearing a dual reporter gene cassette that allowed for dynamic monitoring of HIF-1 signal transduction and tumor cell growth in vitro and in vivo, using optical and nuclear imaging. Tempol, an antioxidant compound, was used to investigate the impact of ROS on cycling hypoxia-mediated HIF-1 activation and tumor progression.Principal Findings
Glioblastoma cells and xenografts were compared under cycling hypoxic and normoxic conditions; upregulation of NOX4 expression and ROS levels were observed under cycling hypoxia in glioblastoma cells and xenografts, concomitant with increased tumor cell growth in vitro and in vivo. However, knockdown of Nox4 inhibited these effects. Moreover, in vivo molecular imaging studies demonstrated that Tempol is a good antioxidant compound for inhibiting cycling hypoxia-mediated ROS production, HIF-1 activation, and tumor growth. Immunofluorescence imaging and flow cytometric analysis for NOX4, HIF-1 activation, and Hoechst 3342 in glioblastoma also revealed high localized NOX4 expression predominantly in potentially cycling hypoxic areas with HIF-1 activation and blood perfusion within the endogenous solid tumor microenvironment.Conclusions
Cycling hypoxia-induced ROS via Nox4 is a critical aspect of cancer biology to consider for therapeutic targeting of cycling hypoxia-promoted HIF-1 activation and tumor progression in GBM. 相似文献13.
Morad Airaki Marina Leterrier Raquel Valderrama Mounira Chaki Juan C. Begara-Morales Juan B. Barroso Luis A. del Río José M. Palma Francisco J. Corpas 《Annals of botany》2015,116(4):679-693
Background and Aims The development of seedlings involves many morphological, physiological and biochemical processes, which are controlled by many factors. Some reactive oxygen and nitrogen species (ROS and RNS, respectively) are implicated as signal molecules in physiological and phytopathological processes. Pepper (Capsicum annuum) is a very important crop and the goal of this work was to provide a framework of the behaviour of the key elements in the metabolism of ROS and RNS in the main organs of pepper during its development.Methods The main seedling organs (roots, hypocotyls and green cotyledons) of pepper seedlings were analysed 7, 10 and 14 d after germination. Activity and gene expression of the main enzymatic antioxidants (catalase, ascorbate–glutathione cycle enzymes), NADP-generating dehydrogenases and S-nitrosoglutathione reductase were determined. Cellular distribution of nitric oxide (·NO), superoxide radical (O2·–) and peroxynitrite (ONOO–) was investigated using confocal laser scanning microscopy.Key Results The metabolism of ROS and RNS during pepper seedling development was highly regulated and showed significant plasticity, which was co-ordinated among the main seedling organs, resulting in correct development. Catalase showed higher activity in the aerial parts of the seedling (hypocotyls and green cotyledons) whereas roots of 7-d-old seedlings contained higher activity of the enzymatic components of the ascorbate glutathione cycle, NADP-isocitrate dehydrogenase and NADP-malic enzyme.Conclusions There is differential regulation of the metabolism of ROS, nitric oxide and NADP dehydrogenases in the different plant organs during seedling development in pepper in the absence of stress. The metabolism of ROS and RNS seems to contribute significantly to plant development since their components are involved directly or indirectly in many metabolic pathways. Thus, specific molecules such as H2O2 and NO have implications for signalling, and their temporal and spatial regulation contributes to the success of seedling establishment. 相似文献
14.
Ferroptosis, a novel type of programmed cell death, is involved in inflammation and oxidation of various human diseases, including diabetic kidney disease. The present study explored the role of high-mobility group box-1 (HMGB1) on the regulation of ferroptosis in mesangial cells in response to high glucose. Compared with healthy control, levels of serum ferritin, lactate dehydrogenase (LDH), reactive oxygen species (ROS), malonaldehyde (MDA), and HMGB1 were significantly elevated in diabetic nephropathy (DN) patients, accompanied with deregulated ferroptosis-related molecules, including long-chain acyl-CoA synthetase 4 (ACSL4), prostaglandin-endoperoxide synthase 2 (PTGS2), NADPH oxidase 1 (NOX1), and glutathione peroxidase 4 (GPX4). In vitro assay revealed that erastin and high glucose both induced ferroptosis in mesangial cells. Suppression of HMGB1 restored cellular proliferation, prevented ROS and LDH generation, decreased ACSL4, PTGS2, and NOX1, and increased GPX4 levels in mesangial cells. Furthermore, nuclear factor E2-related factor 2 (Nrf2) was decreased in DN patients and high glucose-mediated translocation of HMGB1 in mesangial cells. Knockdown of HMGB1 suppressed high glucose-induced activation of TLR4/NF-κB axis and promoted Nrf2 expression as well as its downstream targets including HO-1, NQO-1, GCLC, and GCLM. Collectively, these findings suggest that HMGB1 regulates glucose-induced ferroptosis via Nrf2 pathway in mesangial cells. 相似文献
15.
Kasidit Rattanawong Narumi Koiso Erika Toda Atsuko Kinoshita Mari Tanaka Hiroyuki Tsuji Takashi Okamoto 《The Plant journal : for cell and molecular biology》2021,108(4):1097-1115
Reactive oxygen species (ROS) play essential roles in plant development and environmental stress responses. In this study, ROS dynamics, the glutathione redox status, the expression and subcellular localization of glutathione peroxidases (GPXs), and the effects of inhibitors of ROS-mediated metabolism were investigated along with fertilization and early zygotic embryogenesis in rice (Oryza sativa). Zygotes and early embryos exhibited developmental arrest upon inhibition of ROS production. Egg cells accumulated high ROS levels, and, after fertilization, intracellular ROS levels progressively declined in zygotes in which de novo expression of GPX1 and 3 was observed through upregulation of the genes. In addition to inhibition of GPX activity, depletion of glutathione impeded early embryonic development and led to failure of the zygote to appropriately decrease H2O2 levels. Moreover, through monitoring of the glutathione redox status, the developing zygotes exhibited a progressive glutathione oxidation, which became extremely delayed under inhibited GPX activity. Our results provide insights into the importance of ROS dynamics, GPX antioxidant activity, and glutathione redox metabolism during zygotic/embryonic development. 相似文献
16.
Alexis Díaz-Vegas Cristian A. Campos Ariel Contreras-Ferrat Mariana Casas Sonja Buvinic Enrique Jaimovich Alejandra Espinosa 《PloS one》2015,10(6)
During exercise, skeletal muscle produces reactive oxygen species (ROS) via NADPH oxidase (NOX2) while inducing cellular adaptations associated with contractile activity. The signals involved in this mechanism are still a matter of study. ATP is released from skeletal muscle during electrical stimulation and can autocrinely signal through purinergic receptors; we searched for an influence of this signal in ROS production. The aim of this work was to characterize ROS production induced by electrical stimulation and extracellular ATP. ROS production was measured using two alternative probes; chloromethyl-2,7- dichlorodihydrofluorescein diacetate or electroporation to express the hydrogen peroxide-sensitive protein Hyper. Electrical stimulation (ES) triggered a transient ROS increase in muscle fibers which was mimicked by extracellular ATP and was prevented by both carbenoxolone and suramin; antagonists of pannexin channel and purinergic receptors respectively. In addition, transient ROS increase was prevented by apyrase, an ecto-nucleotidase. MRS2365, a P2Y1 receptor agonist, induced a large signal while UTPyS (P2Y2 agonist) elicited a much smaller signal, similar to the one seen when using ATP plus MRS2179, an antagonist of P2Y1. Protein kinase C (PKC) inhibitors also blocked ES-induced ROS production. Our results indicate that physiological levels of electrical stimulation induce ROS production in skeletal muscle cells through release of extracellular ATP and activation of P2Y1 receptors. Use of selective NOX2 and PKC inhibitors suggests that ROS production induced by ES or extracellular ATP is mediated by NOX2 activated by PKC. 相似文献
17.
18.
Cerebellar granule neurons (CGN) cultured in a 25 mM KCl medium (K25) die apoptotically when they are transferred to a medium
containing 5 mM KCl (K5). It has been previously shown that apoptotic death of CGN induced by K5 is mediated by an increase
in the levels of reactive oxygen species (ROS). ROS may participate in the apoptotic program either as signaling molecules
or as effectors by causing oxidative damage to lipids, DNA and proteins. In this study we evaluated ROS production in CGN
treated with K5 for different periods of time and evaluated a possible correlation between ROS production and oxidation of
DNA proteins and two lipid peroxidation products, conjugated dienes and malondialdehyde. Under these conditions, we found
two episodes of ROS generation, one at an early time (4 h) and another at a later time point (18–24 h). We also identified
two peaks in the formation of conjugated dienes, the initial and transient by-products of lipoperoxidation. The first one
occurred after 4 h of K5 treatment and the other was observed after 18 h, both of them correlated with the formation of ROS.
In contrast, we found significant levels of the late product of lipidperoxidation, malondialdehyde only after 18 h of treatment.
Besides, we did not find significant levels of DNA and protein oxidation products that could be correlated with the observed
ROS production. These results support the idea that ROS produced early by K5 treatment could act primarily as a signal of
the apoptotic cell death and that ROS produced later could be mainly a product of the cell death that could contribute directly
to this process. 相似文献
19.
Balancing inflammatory reactive oxygen species (ROS) production is essential for safely eliminating pathogenic microbes. The newly described protein Negative Regulator of ROS (NRROS) dampens ROS production by restricting NOX2 availability, and thus “cools-off” inflammation.Oxygen-based metabolism was essential for proliferation and diversification of life on Earth but imposed the need to cope with reactive oxygen species (ROS) as an unavoidable aspect of aerobic respiration. Multicellular organisms have evolved a host of mechanisms to utilize ROS to their advantage, thus employing them as mediators of cellular signaling and bactericidal agents against invading pathogens. Excessive ROS production, however, results in oxidative tissue injury, and as recently shown, may participate in the etiology of autoimmunity1. Collateral oxidative cell damage is inseparable from the host innate immunologic responses, underscoring the necessity of regulatory mechanisms that modulate ROS production. A recent publication in Nature has identified a crucial mechanism that balances ROS production and thus modulates the inflammatory response1. Noubade et al.1 describe a protein Negative Regulator of ROS (NRROS, a.k.a. Lrrc33) that interacts with NOX2 (gp91phox) and affects its stability. The NRROS-bound NOX2 in the endoplasmic reticulum (ER) was targeted for proteosomal degradation, and thereby NRROS limited NOX2-derived ROS by impeding NOX2 trafficking to the phagosomal membrane. The authors showed that NRROS and p22phox competed for nascent gp91phox in the ER. Binding to p22phox stabilized gp91phox allowing subsequential trafficking and assembly of the functional NOX2 at the plasma membrane whereas binding to NRROS induced gp91phox trafficking to ER-associated protein degradation (ERAD) pathway (schematically summarized in Figure 1).Open in a separate windowFigure 1Nascent NOX2 interactions in the ER. NOX2 is a major source of superoxide radical anion during phagocytic cell activation. Interaction of nascent NOX2 with p22phox stabilizes NOX2 in the ER and allows for the maturation and trafficking of NOX2 to the plasma membrane where it becomes functional. NRROS is a newly identified NOX2-binding partner in the ER. Binding of NOX2 by NRROS accelerates NOX2 degradation, and thereby modulates ROS production by reducing NOX2 level.NOX2 is a major source of superoxide radicals produced by phagocytic cells during the oxidative burst. Thus, an important discovery made is that NRROS is downregulated in response to priming by pro-inflammatory stimuli including lipopolysaccharide and type II interferon. NRROS deficiency resulting from exposure to these stimuli caused the cells to respond to subsequent inflammatory challenges with an appropriately robust ROS burst required for killing bacteria and viruses. The kinetics of ROS production in stimulated NRROS-deficient versus wild-type cells was identical, suggesting that NRROS functions to establish the upper limit in the achievable levels of ROS production following phagocyte activation. These observations provide important insights into the widely recognized but poorly understood amplifying effect of basal inflammation on the subsequent activation of innate immune responses. The implication is that chronic exposure to low levels of pro-inflammatory mediators can exacerbate the availability of functional NOX2 by chronically repressing NRROS. In support of this idea, NRROS-knockout mice were better equipped to fight infection by L. monocytogenes and survived better in contrast to wild-type animals, consistent with the known effect of ROS in promoting pathogen killing. On the flipside, however, elimination of a central repressive mechanism limiting NOX2-derived ROS induced lethal encephalomyelitis (EAE) with rapid deterioration of the central nervous system in immunized NRROS-knockout mice1. A possible clinically relevant interpretation of this observation is that while severe oxidative stress facilitates elimination of pathogenic organisms, it can also overwhelm the host''s ability to clear oxidized biomolecules which may trigger the onset of autoimmunity. The findings raise the intriguing question whether dysregulated ROS production during successive acute inflammatory events is conducive to the pathogenesis of autoimmune disorders. This concept also resonates with some earlier studies highlighting the effects of environment-induced chronic systemic inflammation and the propensity of mammals to develop degenerative neurological disorders2,3. For example, it was observed that exacerbated NOX2 activity underlies microglia-mediated neurotoxicity that can lead to Parkinson''s and Alzheimer''s diseases2. It was previously theorized that chronic low-grade inflammatory states promoted by exposure to environmental toxicants (air pollution, pesticides, etc.) primes microglia (the macrophages of the brain) to produce exaggerated amounts of ROS generated largely by NOX23. As demonstrated by studies testing this hypothesis, microglia from animals exposed to diesel exhaust particles produced robust bursts of ROS when subsequently challenged with lipopolysaccharides2. Levels of ROS produced by primed microglia were demonstrated to be neurotoxic to dopaminergic neurons and induce neurodegeneration in a mouse model4. Other studies showed that feedback signaling by NOX2-derived ROS limits TNFα and interleukin-6 expression by activated macrophages, which alleviated acute inflammatory lung injury5. The production of ROS by specialized systems is currently considered to be limited by cofactor and O2 availability6. The identification of NRROS and its function indicates that on the contrary specific mechanisms exist to dynamically regulate the levels of ROS. Taken together, these studies suggest that budgeting ROS production is indispensable for safe interactions of the host with the environment. They also suggest that exacerbated ROS production leads to production of pseudo-antigens and oxidized biomolecules whose clearance may be the rate-limiting factor, and hence the necessity to limit their production by dampening ROS generation. This provocative hypothesis is consistent with the finding of higher levels of malonaldehyde (MDA)-adduct proteins after immunization in NRROS-deficient mice1. Another bit of evidence supporting this idea is the finding that administration of ROS scavengers after the onset of autoimmune EAE reduced the lethality of immunized NRROS-knockout mice back to wild-type control levels1. Although the levels of oxidized proteins were not assessed after ROS scavenger treatment, the near abrogation of autoimmune EAE by ROS scavengers administered after the onset of EAE indicates that resolution of ongoing oxidative stress (or the clearance of oxidized pseudo-antigens) is sufficient to prevent further neurologic deterioration.The role of ROS as mediators of tissue damage has been established (see reviews7,8). To date, much less is understood about the role of ROS in regulating the initiation, intensity, localization, and resolution of the inflammatory process. As the redox sensitivity of different signaling cascades varies, the concentrations and kinetics of ROS production are likely to shape specific inflammatory programs. Dysregulated ROS generation as shown by this study1 is likely to cause detrimental effects produced by inflammation gone awry. Addressing questions related to the integration of redox signaling in inflammation and temporal control of differential ROS fluxes will further our understanding about the role of ROS in modulating inflammation and will provide exciting opportunities for therapeutic interventions. 相似文献
20.
Paula A. Belinky Nufar Flikshtein Sergey Lechenko Shimon Gepstein Carlos G. Dosoretz 《Applied microbiology》2003,69(11):6500-6506
We studied oxidative stress in lignin peroxidase (LIP)-producing cultures (cultures flushed with pure O2) of Phanerochaete chrysosporium by comparing levels of reactive oxygen species (ROS), cumulative oxidative damage, and antioxidant enzymes with those found in non-LIP-producing cultures (cultures grown with free exchange of atmospheric air [control cultures]). A significant increase in the intracellular peroxide concentration and the degree of oxidative damage to macromolecules, e.g., DNA, lipids, and proteins, was observed when the fungus was exposed to pure O2 gas. The specific activities of manganese superoxide dismutase, catalase, glutathione reductase, and glutathione peroxidase and the consumption of glutathione were all higher in cultures exposed to pure O2 (oxygenated cultures) than in cultures grown with atmospheric air. Significantly higher gene expression of the LIP-H2 isozyme occurred in the oxygenated cultures. A hydroxyl radical scavenger, dimethyl sulfoxide (50 mM), added to the culture every 12 h, completely abolished LIP expression at the mRNA and protein levels. This effect was confirmed by in situ generation of hydroxyl radicals via the Fenton reaction, which significantly enhanced LIP expression. The level of intracellular cyclic AMP (cAMP) was correlated with the starvation conditions regardless of the oxygenation regimen applied, and similar cAMP levels were obtained at high O2 concentrations and in cultures grown with atmospheric air. These results suggest that even though cAMP is a prerequisite for LIP expression, high levels of ROS, preferentially hydroxyl radicals, are required to trigger LIP synthesis. Thus, the induction of LIP expression by O2 is at least partially mediated by the intracellular ROS. 相似文献