Exogenous nitric oxide protect cucumber roots against oxidative stress induced by salt stress.

Mitochondria are subcellular organelles with an essentially oxidative type of metabolism. The production of reactive oxygen species (ROS) in it increases under stress conditions and causes oxidative damage. In the present study, effects of exogenous sodium nitroprusside (SNP), a nitric oxide (NO) donor, on both the ROS metabolism in mitochondria and functions of plasma membrane (PM) and tonoplast were studied in cucumber seedlings treated with 100mM NaCl. NaCl treatment induced significant accumulation of H(2)O(2) and led to serious lipid peroxidation in cucumber mitochondria, and the application of 50muM SNP stimulated ROS-scavenging enzymes and reduced accumulation of H(2)O(2) in mitochondria of cucumber roots induced by NaCl. As a result, lipid peroxidation of mitochondria decreased. Further investigation showed that application of SNP alleviated the inhibition of H(+)-ATPase and H(+)-PPase in PM and/or tonoplast by NaCl. While application of sodium ferrocyanide (an analog of SNP that does not release NO) did not show the effect of SNP, furthermore, the effects of SNP were reverted by addition of hemoglobin (a NO scavenger).     

Differential protection against oxidative stress and nitric oxide overproduction in cardiovascular and pulmonary systems by propofol during endotoxemia

Background  

Both overproduction of nitric oxide (NO) and oxidative injury of cardiovascular and pulmonary systems contribute to fatal cardiovascular depression during endotoxemia. We investigated in the present study the relative contribution of oxidative stress and NO to cardiovascular depression during different stages of endotoxemia, and delineated their roles in cardiovascular protective effects of a commonly used anesthetic propofol during endotoxemia.     

Inducible nitric oxide synthase and cyclooxgenase-2 mediate protection of hydrogen peroxide preconditioning against apoptosis induced by oxidative stress in PC12 cells

The induction of inducible nitric oxide synthase (iNOS) in response to different stress is associated with simultaneous induction of cyclooxygenase-2 (COX-2) in various cell types. Both iNOS and COX-2 have been reported to mediate the late phase of cardioprotection induced by different preconditioning. However, whether both iNOS and COX-2 are mediators in the neuroprotection induced by preconditioning with hydrogen peroxide (H(2)O(2)) at low concentration is unknown. In this study, using the neurosecretory cell line-PC12 cells to set up the model of neuroprotection of preconditioning with H(2)O(2) against apoptosis, we first investigate what changes in expression of iNOS and COX-2 appear during H(2)O(2) preconditioning, then determine if both iNOS inhibitor and COX-2 inhibitor interfere with the neuroprotection elicited by preconditioning with H(2)O(2). We found that preconditioning with H(2)O(2) at 10 microM significantly protected PC12 cells against apoptosis induced by lethal H(2)O(2) (50 microM) and increased the expression of iNOS and COX-2 and that selective iNOS inhibitor, aminoguanidine (AG) and COX-2 inhibitor, NS-398 obviously blocked the protective effects induced by preconditioning with 10 microM H(2)O(2). The results of this study suggest that both iNOS and COX-2 are mediators of the neuroprotection induced by preconditioning with oxidative stress (H(2)O(2) at low concentration) in PC12 cells.     

Application of the nitric oxide donor SNAP to cardiomyocytes in culture provides protection against oxidative stress.

Multiple data indicates that nitric oxide (NO) donors retain immediate protective effects against different disturbances in cardiovascular system. The aim of the present study was to investigate delayed effects of nitric oxide donor S-nitroso-N-acetyl-l,l-penicillamine (SNAP) application in cardiac H9c2 cell line. Cardiomyocytes were treated with SNAP for 2h followed by 24h wash with fresh growth medium. The concentration curve was constructed in range from 0.5 to 2mM, toxicity was observed at 2mM concentration of SNAP. For the study of SNAP-induced protection against t-butyl hydroperoxide-induced oxidative injury 1mM concentration was used. Cell viability was assessed by MTT reductase activity assay; mitochondrial transmembrane potential (mdeltapsi) was measured by flow cytometry with fluorescent dye DiOC(6). Synthesis of heat-shock proteins (hsps) was analyzed by Western blot. Analysis of the cell viability and mdeltapsi reflected delayed protective effect of 1mM SNAP application against oxidative injury. SNAP in 1mM concentration caused 70% induction of hsp75 synthesis in cardiomyocytes. However, the other analyzed hsps (hsp70, hsp27, hsp60, hsp10, and CyP A) did not display any significant induction after incubation with SNAP. Present work demonstrates that the NO donor SNAP causes delayed protection against oxidative stress in H9c2 cardiomyocyte cell line, reflected in cell viability increase and preservation of the mdeltapsi. We suppose the major pathway for the development of SNAP-induced protection is through mitochondria. Induction of hsp75 expression following SNAP pretreatment is one possible way to explanation the mechanisms of this protection.     

Taurine protection of PC12 cells against endoplasmic reticulum stress induced by oxidative stress

Background

Taurine is a free amino acid present in high concentrations in a variety of organs of mammalians. As an antioxidant, taurine has been found to protect cells against oxidative stress, but the underlying mechanism is still unclear.

Methods

In this report, we present evidence to support the conclusion that taurine exerts a protective function against endoplasmic reticulum (ER) stress induced by H₂O₂ in PC 12 cells. Oxidative stress was introduced by exposure of PC 12 cells to 250 uM H₂O₂ for 4 hours.

Results

It was found that the cell viability of PC 12 cells decreased with an increase of H₂O₂ concentration ranging from approximately 76% cell viability at 100 uM H₂O₂ down to 18% at 500 uM H₂O₂. At 250 uM H₂O₂, cell viability was restored to 80% by taurine at 25 mM. Furthermore, H₂O₂ treatment also caused a marked reduction in the expression of Bcl-2 while no significant change of Bax was observed. Treatment with taurine restored the reduced expression of Bcl-2 close to the control level without any obvious effect on Bax. Furthermore, taurine was also found to suppress up-regulation of GRP78, GADD153/CHOP and Bim induced by H₂O₂, suggesting that taurine may also exert a protective function against oxidative stress by reducing the ER stress.

Conclusion

In summary, taurine was shown to protect PC12 cells against oxidative stress induced by H₂O₂. ER stress was induced by oxidative stress and can be suppressed by taurine.

     

Cellular protection of morin against the oxidative stress induced by hydrogen peroxide

Flavonoids are a class of secondary metabolites abundantly found in fruits and vegetables. In addition, flavonoids have been reported as potent antioxidants with beneficial effects against oxidative stress-related diseases such as cancer, aging, and diabetes. The present study was carried out to investigate the cytoprotective effects of morin (2′,3,4′,5,7-pentahydroxyflavone), a member of the flavonoid group, against hydrogen peroxide (H₂O₂)-induced DNA and lipid damage. Morin was found to prevent the cellular DNA damage induced by H₂O₂ treatment, which is shown by the inhibition of 8-hydroxy-2′-deoxyguanosine (8-OHdG) formation (a modified form of DNA base), inhibition of comet tail (a form of DNA strand breakage), and decrease of nuclear phospho histone H2A.X expression (a marker for DNA strand breakage). In addition, morin inhibited membrane lipid peroxidation, which is detected by inhibition of thiobarbituric acid reactive substance (TBARS) formation. Morin was found to scavenge the intracellular reactive oxygen species (ROS) generated by H₂O₂ treatment in cells, which is detected by a spectrofluorometer, flow cytometry, and confocal microscopy after staining of 2′,7′-dichlorodihydrofluorescein diacetate (DCF-DA). Morin also induces an increase in the activity of catalase and protein expression. The results of this study suggest that morin protects cells from H₂O₂-induced damage by inhibiting ROS generation and by inducing catalase activation.     

Hydrogen sulfide counteracts chlorophyll loss in sweetpotato seedling leaves and alleviates oxidative damage against osmotic stress

Sawgrass (Cladium jamaicense) is the predominant plant and vegetation community in the Florida Everglades. Germination of sawgrass seeds in the laboratory or nursery has been difficult and problematic, yet little is known about the physiological mechanistic regulation of the sawgrass seed germination process. In the present study, we examined the factors and mechanisms that influence sawgrass seed germination. We found that removal of seed husk and bracts, pre-soaking with bleach (hypochlorite), breaking the seed coat, or combinations of these treatments promoted the rate and success of germination, whereas presence of seed-encasing structures or treatment with husk/bract extract inhibited germination. We further detected the presence of abscisic acid (ABA) in the husk and bract. Experiments with ABA and gibberellin biosynthesis inhibitors fluridone and tetcyclacis suggested that ABA already presented in the pre-imbibed seeds, and not derived through post-dormancy de novo synthesis, contributed to the inhibition of seed germination. Examination of bleach and mechanical treatments indicated the physical barrier presented by the seed-encasing structures provided additional mechanism for the long-term delay of seed germination. Based on the results of this study and others, we discussed the implications of sawgrass seed dormancy and germination in relation to its natural habitat and proposed a hypothesis that the protracted seed dormancy in sawgrass offered an adaptive advantage in the pre-anthropogenic Everglades environment, but may become a liability in the current man-managed Everglades water system.     

Protective effect of nitric oxide against oxidative stress under ultraviolet-B radiation.

The response of bean leaves to UV-B radiation was extensively investigated. UV-B radiation caused increase of ion leakage, loss of chlorophyll, and decrease of the maximum efficiency of PSII photochemistry (Fv/Fm) and the quantum yield of PSII electron transport (PhiPSII) of bean leaves. H2O2 contents and the extent of thylakoid membrane protein oxidation increased, indicated by the decrease of thiol contents and the increase of carbonyl contents with the duration of UV-B radiation. Addition of sodium nitroprusside, a nitric oxide (NO) donor, can partially alleviate UV-B induced decrease of chlorophyll contents, Fv/Fm and PhiPSII. Moreover, the oxidative damage to the thylakoid membrane was alleviated by NO. The potassium salt of 2-(4-carboxyphenyl)-4,4,5,5-tetramethylimidazoline-1-oxyl-3-oxide, a specific NO scavenger, arrested NO mediated protective effects against UV-B induced oxidative damage. Incubation of thylakoid membrane with increasing H2O2 concentrations showed a progressive enhancement in carbonyl contents. H2O2 contents were decreased in the presence of NO under UV-B radiation through increased activities of superoxide dismutases, ascorbate peroxidases, and catalases. Taken together, the results suggest that NO can effectively protect plants from UV-B damage mostly probably mediated by enhanced activities of antioxidant enzymes.     

Hepatocellular oxidative DNA injury induced by macrophage-derived nitric oxide

Previous studies have indicated that splenic macrophages migrate into the liver and play a role in endotoxin-induced hepatic damage. The present study was designed to elucidate the mechanisms of hepatocyte injury induced by activated splenic macrophages, focusing especially on endogenously released NO and oxidative DNA alterations in hepatocytes. Splenic macrophages isolated from Wistar rats were incubated with either lipopolysaccharide (LPS) or interferon-gamma (IFN-gamma) and cocultured with hepatocytes. Nitrite and nitrate levels in the culture medium were measured, and inducible-type NO synthase (iNOS) and nitrotyrosine were determined by immunofluorescence staining. The ratio of 8-hydroxy-deoxyguanosine (8-OH-dG) to deoxyguanosine (dG) was measured by high-performance liquid chromatography, and single-stranded DNA in hepatocytes was detected with acridine orange. NO release and nitrotyrosine expression in hepatocytes increased after 8 h of coculture with activated macrophages, and this coculture also induced increases in the 8-OH-dG/dG ratio and single-stranded DNA in the hepatocytes. These alterations were attenuated by superoxide dismutase (SOD) and NO synthesis inhibitors. A similar pattern of alterations was observed in hepatocytes incubated with SIN-1, and these changes were also prevented by SOD. These results suggest that activated macrophage-derived NO and its oxidative metabolite, peroxynitrite, play key roles in hepatocyte injury during inflammation, and cause subsequent DNA damage in surviving hepatocytes.     

NO对盐胁迫下苜蓿根系生长抑制及氧化损伤的缓解效应

以"甘农4号"苜蓿品种为材料,采用水培法,用NO供体硝普钠(SNP)、硝普钠类似物亚铁氰化钠(不产生NO)、NO特异清除剂c-PTIO、一氧化氮合酶(NOS)活性抑制剂N-硝基-L-精氨酸甲脂(L-NAME)、硝酸还原酶(NR)活性抑制剂钨酸盐处理苜蓿植株,研究NO对盐胁迫下苜蓿幼苗根系生长、根系活力、根系中渗透调节物质、膜脂过氧化、活性氧含量及抗氧化酶活性等的影响,探讨NO调控苜蓿幼苗根系耐盐性的生理机制。结果表明:盐胁迫下SNP处理提高了根系活力,促进了苜蓿幼苗根系生长,降低游离脯氨酸含量,促进可溶性蛋白含量增加;增强超氧化物歧化酶(SOD)、过氧化氢酶(CAT)、愈创木酚过氧化物酶(GPX)、抗坏血酸过氧化物酶(APX)和谷胱甘肽还原酶(GR)活性,提高还原型抗坏血酸(As A)和还原型谷胱甘肽(GSH)含量,降低过氧化氢(H2O2)、羟自由基(OH·)含量、超氧阴离子(O·-2)产生速率和膜脂过氧化产物丙二醛(MDA)含量;同时,SNP处理显著促进了苜蓿幼苗根系内源NO的积累。NO供体SNP的类似物亚铁氰化钠对盐胁迫下苜蓿根系各项生理生化指标无明显影响;盐胁迫下添加c-PTIO、L-NAME和钨酸盐进一步降低了苜蓿幼苗根系活力和根系生长,抑制了根系抗氧化系统活性,加剧了根系膜脂过氧化作用,降低了内源NO积累,添加SNP则能缓解该抑制效应;表明外源SNP处理能明显缓解盐胁迫对苜蓿幼苗根系生长的抑制和氧化损伤,且通过NOS和NR途径产生的内源NO也可能在苜蓿根系适应盐胁迫的调节中起关键作用;该研究结果为苜蓿耐盐机制及NO在苜蓿耐盐育种、化学调控和盐碱地栽培利用等提供了理论依据。     

Exogenous nitric oxide improves seed germination in wheat against mitochondrial oxidative damage induced by high salinity

Effects of exogenous nitric oxide (NO) on starch degradation, oxidation in mitochondria and K⁺/Na⁺ accumulation during seed germination of wheat were investigated under a high salinity level. Seeds of winter wheat (Triticum aestivum L., cv. Huaimai 17) were pre-soaked with 0 mM or 0.1 mM of sodium nitroprusside (SNP, as nitric oxide donor) for 20 h just before germination under 300 mM NaCl. At 300 mM NaCl, exogenous NO increased germination rate and weights of coleoptile and radicle, but decreased seed weight. Exogenous NO also enhanced seed respiration rate and ATP synthesis. In addition, seed starch content decreased while soluble sugar content increased by exogenous NO pre-treatment, which was in accordance with the improved amylase activities in the germinating seeds. Exogenous NO increased the activities of superoxide dismutase (SOD, EC 1.15.1.1) and catalase (CAT, EC 1.11.1.6); whereas decreased the contents of malondialdehyde (MDA) and hydrogen peroxide (H₂O₂), and superoxide anions (O₂^??) release rate in the mitochondria. Exogenous NO also decreased Na⁺ concentration while increased K⁺ concentration in the seeds thereby maintained a balance between K⁺ and Na⁺ during germination under salt stress. It is concluded that exogenous NO treatment on wheat seeds may be a good option to improve seed germination and crop establishment under saline conditions.     

Increased stability of Bcl-2 in HSP70-mediated protection against apoptosis induced by oxidative stress

We have previously shown that heat shock protein 70 (HSP70) markedly inhibits H₂O₂-induced apoptosis in mouse C2C12 myogenic cells by reducing the release of Smac. However, the molecular mechanism by which HSP70 interferes with Smac release during oxidative stress-induced apoptosis is not understood. In the current study, we showed that HSP70 increased the stability of Bcl-2 during oxidative stress. An antisense phosphorothioate oligonucleotide against Bcl-2 caused selective inhibition of Bcl-2 protein expression induced by HSP70 and significantly attenuated HSP70-mediated cell protection against H₂O₂-induced release of Smac and apoptosis. Taken together, our results indicate that there are important relationships among HSP70, Bcl-2, release of Smac, and induction of apoptosis by oxidative stress.     

Overexpression of SsCHLAPXs confers protection against oxidative stress induced by high light in transgenic Arabidopsis thaliana

To evaluate the physiological importance of chloroplastic ascorbate peroxidase (CHLAPX) in the reactive oxygen species (ROS)‐scavenging system of a euhalophyte, we cloned the CHLAPX of Suaeda salsa (SsCHLAPX) encoding stromal APX (sAPX) and thylakoid‐bound APX. The stromal APX of S. salsa (Ss.sAPX) cDNA consists of 1726 nucleotides including an 1137‐bp open reading frame (ORF) and encodes 378 amino acids. The thylakoid‐bound APX of S. salsa (Ss.tAPX) cDNA consists of 1561 nucleotides, including a 1284‐bp ORF, and encodes 427 amino acids. The N‐terminal 378 amino acids of Ss.sAPX are identical with those of Ss.tAPX, whereas the C‐terminal 49 amino acids differ. Arabidopsis thaliana lines overexpressing Ss.sAPX and Ss.tAPX were constructed using Agrobacterium tumefaciens transformation methods. Under high light (1000 µmol m^?2 s^?1), malondialdehyde (MDA) content was lower in transgenic plants than in the wild type. Under high light, Fv/Fm and chlorophyll contents of both overexpressing lines and the wild type declined but were significantly higher in the overexpressing lines than in the wild type. The activities of APX (EC 1.11.1.11), catalase (CAT 1.11.1.6) and superoxide dismutase (SOD EC 1.15.1.1) were higher in the overexpressing lines than in the wild type. The transgenic plants showed increased tolerance to oxidative stress caused by high light. These results suggest that SsCHLAPX plays an important role in scavenging ROS in chloroplasts under stress conditions such as high light.     

Garlic provides protection to mice heart against isoproterenol-induced oxidative damage: role of nitric oxide

Garlic has been widely recognized as a cardioprotective agent. However, the molecular mechanism of its cardioprotective effects is not well established. Here we hypothesized that aqueous garlic homogenate may mediate cardioprotection via nitric oxide (NO). Mice were fed with saline and aqueous garlic homogenate (250 and 500 mgkg(-1)day(-1) orally) for 30 days. In another set of experiment, mice were pre-treated with saline, aqueous garlic homogenate (AGH) (250 mgkg(-1)day(-1) for 30 days), and AGH (30 days) along with L-NAME (20 mgkg(-1)day(-1) i.p. for last 7 days) before inducing acute myocardial infarction by isoproterenol (s.c. injection of isoproterenol 150 mgkg(-1)day(-1) for 2 days) and sacrificed after 48 h. Dose dependent increase in serum NO level was observed after garlic 250 and 500 mgkg(-1) dose feeding. While no change in serum SGPT and SGOT level, a significant decrease in serum LDH level was observed after garlic feeding. Garlic-induced NO formation was further confirmed in human aortic endothelial cells (HAEC). Administration of isoproterenol caused a significant decrease in endogenous antioxidants i.e., myocardial catalase, GSH and GPx activity, and mitochondrial enzyme activities like citrate synthase and β hydroxyacyl CoA dehydrogenase. All those deleterious cardiac changes induced by isoproterenol were significantly attenuated by garlic homogenate. However this beneficial effect of garlic was blunted when garlic was administered with L-NAME, a nonspecific inhibitor of nitric oxide synthase (NOS). Further, a significant increase in myocardial TBARS and decrease in total antioxidant activity was observed in L-NAME treated group compared to isoproterenol treated group. Administration of L-NAME in mice from control group lowered serum and cardiac NO levels without any change of oxidative stress parameters. In conclusion, our study provides novel evidence that garlic homogenate is protective in myocardial infarction via NO-signaling pathway in mice.     

Diazoxide triggers cardioprotection against apoptosis induced by oxidative stress

Although mitochondrial ATP-sensitive potassium (mitoK(ATP)) channels have been reported to reduce the extent of apoptosis, the critical timing of mitoK(ATP) channel opening required to protect myocytes against apoptosis remains unclear. In the present study, we examined whether the mitoK(ATP) channel serves as a trigger of cardioprotection against apoptosis induced by oxidative stress. Apoptosis of cultured neonatal rat cardiomyocytes was determined by flow cytometry (light scatter and propidium iodide/annexin V-FITC fluorescence) and by nuclear staining with Hoechst 33342. Mitochondrial membrane potential (DeltaPsi) was measured by flow cytometry of cells stained with rhodamine-123 (Rh-123). Exposure to H(2)O(2) (500 microM) induced apoptosis, and the percentage of apoptotic cells increased progressively and peaked at 2 h. This H(2)O(2)-induced apoptosis was associated with the loss of DeltaPsi, and the time course of decrease in Rh-123 fluorescence paralleled that of apoptosis. Pretreatment of cardiomyocytes with diazoxide (100 microM), a putative mitoK(ATP) channel opener, for 30 min before exposure to H(2)O(2) elicited transient and mild depolarization of DeltaPsi and consequently suppressed both apoptosis and DeltaPsi loss after 2-h exposure to H(2)O(2). These protective effects of diazoxide were abrogated by the mitoK(ATP) channel blocker 5-hydroxydecanoate (500 microM) but not by the sarcolemmal K(ATP) channel blocker HMR-1098 (30 microM). Our results suggest for the first time that diazoxide-induced opening of mitoK(ATP) channels triggers cardioprotection against apoptosis induced by oxidative stress in rat cardiomyocytes.     

Role of nitric oxide dependence on nitric oxide synthase-like activity in the water stress signaling of maize seedling

Nitric oxide (NO) has been known as an important signal in plant antioxidative defense but its production and roles in water stress are less known. The present study investigated whether NO dependence on a NO synthase-lika (NOS) activity is involved in the signaling of drought-induced protective responses in maize seedlings. NOS activity, rate of NO release and drought responses were analyzed when NO donor sodium nitroprusside (SNP), NO scavenger c-PTIO (2-(4-carboxyphenyl)-4,4,5,5-tetramathylimidazoline-1-oxyl-3-oxide) and NOS inhibitor L-NAME (NG-nitro-L-arginine methyl ester) were applied to both detached maize leaves and whole plants. Both NOS activity and the rate of NO release increased substantially under dehydration stress. The high NOS activity induced by c-PTIO as NO scavenger and NO accumulation Inhibited by NOS inhibitor L-NAME In dehydration-treated maize seedlings Indicated that most NO production under water deficit stress may be generated from NOS-like activity. After dehydration stress for 3 h, detached maize leaves pretreated with NO donor SNP maintained more water content than that of control leaves pretreated with water. This result was consistent with the decrease in the transpiration rate of SNP-treated leaves subjected to drought treatment for 3 h. Membrane permeability, a cell injury index, was lower in SNP-trested maize leaves under dehydration stress for 4 h when compared with the control leaves. Also, superoxide dismutsse (SOD) activity of SNP combined drought treatment maize leaves was higher than that of drought treatment alone, indicating that exogenous NO treatment alleviated the water loss and oxidative damage of maize leaves under water deficit stress. When c-PTIO as a specific NO scavenger was applied, the effects of applied SNP were overridden. Treatment with L-NAME on leaves also led to higher membrane permeability, higher transpiration rate and lower SOD activities than those of control leaves, indicating that NOS-like activity was involved in the antioxidative defense under water stress. These results suggested that NO dependence on NOS-like activity serves as a signaling component in the induction of protective responses and is associated with drought tolerance in maize seedlings.     

Mitochondrial nitric oxide synthase, oxidative stress and apoptosis

Nitric oxide (NO) exerts a wide range of its biological properties via its interaction with mitochondria. By competing with O(2), physiologically relevant concentrations of NO reversibly inhibit cytochrome oxidase and decrease O(2) consumption, in a manner resembling a pharmacological competitive antagonism. The inhibition regulates many cellular functions, by e.g., regulating the synthesis of ATP and the formation of mitochondrial transmembrane potential (Delta Psi). NO regulates the oxygen consumption of both the NO-producing and the neighboring cells; thus, it can serve as autoregulator and paracrine modulator of the respiration. On the other hand, NO reacts avidly with superoxide anion (O(2)(-)) to produce the powerful oxidizing agent, peroxynitrite (ONOO(-)) which affects mitochondrial functions mostly in an irreversible manner. How mitochondria and cells harmonize the reversible effects of NO versus the irreversible effects of ONOO(-) will be discussed in this review article. The exciting recent finding of mitochondrial NO synthase will also be discussed.     

HSP72 as a complementary protection against oxidative stress induced by exercise in the soleus muscle of rats

Given the potential of reactive oxygen species to damage intracellular proteins during subsequent bouts of muscle contractions, it was suggested that, when this production exceeds the antioxidant capacity, the preexisting antioxidant pathways may be complemented by the synthesis of the defense mechanism represented by heat shock proteins (HSPs), stress proteins with the function of repair and maintaining protein folding. To test this hypothesis, we analyzed reactive carbonyl derivatives in plasma and the expression of HSP72 and activities of enzymes from the oxidative and antioxidant defense systems in the soleus muscle of sedentary rats and rats trained by two protocols: continuous and intermittent. We analyzed all three groups at rest and 2 h after acute exercise. After 8 wk of training, the animals from both groups clearly demonstrated higher resistance to exercise. Both trained groups showed significantly higher citrate synthase, catalase, and glutathione reductase activities than the control group (P < 0.01). After acute exercise, catalase and glutathione reductase activities significantly decreased (P < 0.01) and plasma reactive carbonyl derivatives significantly increased (P < 0.05) in the sedentary group, suggesting an oxidative-stress condition as responsible for exhaustion in this group. Finally, after acute exercise, the induction of HSP72 expression occurred only in the sedentary group, suggesting that HSP72 acts as a complementary protective mechanism in exercise-induced oxidative stress.     

Inducible nitric oxide synthase protection against coxsackievirus pancreatitis.

Coxsackievirus infection causes myocarditis and pancreatitis in humans. In certain strains of mice, Coxsackievirus causes a severe pancreatitis. We explored the role of NO in the host immune response to viral pancreatitis. Coxsackievirus replicates to higher titers in mice lacking NO synthase 2 (NOS2) than in wild-type mice, with particularly high viral titers and viral RNA levels in the pancreas. Mice lacking NOS have a severe, necrotizing pancreatitis, with elevated pancreatic enzymes in the blood and necrotic acinar cells. Lack of NOS2 leads to a rapid increase in the mortality of infected mice. Thus, NOS2 is a critical component in the immune response to Coxsackievirus infection.