Serum Reactive Oxygen Species Research Articles

Associaiton of irisin and vaspin with clinical presentations of metabolic syndrome in patients with type 2 diabetes mellitus

Objective To explore the changes in serum concentrations of irisin, vaspin and reactive oxygen species (ROS) in patients with type 2 diabetes mellitus (T2DM) and metabolic syndrome (MS), and to investigated the correlation of irisin and vaspin with clinical parameters of MS. Methods A total of 260 T2DM patients were enrolled. Age and gender were recorded, anthropometrics, biochemical parameters, and levels of irisin, vaspin and ROS in fasting serum were measured, and homeostatic model assessment of insulin resistance (HOMA-IR) calculated. Wilcoxon rank sum test, correlation analysis, Logistic regression analysis, multiple linear regression analysis and receiver operating characteristic (ROC) curve analysis were performed. Results Compared to T2DM patients without MS, T2DM patients with MS had lower serum level of irisin [male: 112.81(86.96-191.84)μg/L vs. 156.23(110.61-225.97)μg/L, female: 141.09(77.52-175.55)μg/L vs. 172.15(95.69-240.37)μg/L, P<0.01], higher levels of vaspin and ROS [male: 1.13(0.95-1.38)μg/L vs. 0.36(0.21-0.82)μg/L, 1 540(1 250-1 860)kU/L vs. 1 020(920-1 350)kU/L; female: 1.52(1.13-1.80)μg/L vs. 0.51(0.47-1.08)μg/L, 1 650(1 320-1 940)kU/Lvs. 1 120(980-1 420)kU/L, P<0.01]. In the T2DM patients, serum irisin level was negatively correlated with vaspin (r=-0.382, P<0.01) and ROS (r=-0.410, P<0.01), while vaspin was positively correlated with ROS (r=0.400, P<0.01). Multiple linear regression analyses showed that irisin was significantly correlated with body mass index (BMI), waist circumference and triglyceride, while vaspin was correlated with gender, BMI, and waist circumference (all P<0.05). Logistic regression analysis revealed that irisin, vaspin and ROS were all associated with MS (OR=0.77, 95% CI 0.608-0.978; OR=1.39, 95% CI 1.252-1.539; OR=1.38, 95% CI 1.112-1.718, all P<0.05). ROC analysis demonstrated that irisin and vaspin had significant area under the curve (AUC=0.931, P<0.01; AUC=0.777, P<0.01) for the prediction of MS. Conclusions Serum irisin level was significantly decreased, while vaspin and ROS were significantly increased in T2DM patients with MS. Irisin and vaspin were associated with clinical presentations of MS, suggesting that irisin and vaspin might be valuable predictors of MS. Key words: Metabolic syndrome; Diabetes mellitus; Reactive oxygen species; Irisin; Vaspin

Enhanced-autophagy by exenatide mitigates doxorubicin-induced cardiotoxicity

ObjectivesExenatide is a glucagon-like peptide-1 analogue that mitigates myocardial injury caused by ischemia-reperfusion injury via the survival signaling pathway. We hypothesized that exenatide would provide a protective effect in doxorubicin-induced cardiotoxicity. MethodsH9c2 cardiomyocytes were pre-treated with exenatide followed by doxorubicin (DOX), and cell viability and intracellular reactive oxygen species (ROS) were subsequently measured. In order to determine the role of autophagy, we performed western blot as well as TUNEL and autophagosome staining. Additionally, rats were treated with exenatide 1h prior to every DOX treatment. Left ventricular (LV) function and performance were then assessed by echocardiography. Myocardial and serum ROS was measured with DHE fluorescence and ROS/RNS assay. ResultsDOX-induced caspase-3 activation decreased after pre-treatment with exenatide both in vivo and in vitro. Oxidative stress was attenuated by exenatide in H9c2 cells, as well as in cardiac tissue and serum. The number of autophagosomes and autophagic markers were further increased by exenatide in the DOX-treated H9c2 cells, which mediated AMPK activation. Suppression of the autophagosome abolished exenatide-induced anti-apoptotic effect. Echocardiography showed that pre-treatment with exenatide significantly improved LV dysfunction that is induced by DOX treatment. Exenatide inhibits the DOX-induced production of intracellular ROS and apoptosis in the myocardium. The autophagic markers increased in exenatide pre-treated cardiac tissue. ConclusionExenatide reduces DOX-induced apoptosis of cardiomyocytes by upregulating autophagy and improving cardiac dysfunction. These novel results highlight the therapeutic potential of exenatide to prevent doxorubicin cardiotoxicity.

The Nrf2/HO-1 Pathway Mediates the Antagonist Effect of L-Arginine On Renal Ischemia/Reperfusion Injury in Rats

Background/Aims: Ischemia/reperfusion (I/R) is the most common cause of acute renal injury. I/R-induced oxidative stress is involved in the development of acute renal injury, which can be reversed by supplementation with L-arginine, a precursor of nitric oxide (NO). This study was conducted to evaluate alterations in the expression of transcription factors [nuclear factor kappa B (NF-κB), nuclear factor-E2-related factor-2 (Nrf2), and heme oxygenase 1 (HO-1)] and heat shock protein 70 (HSP70) in the kidney of I/R-induced injury rats. Methods: Sprague-Dawley (SD) rats were subjected to bilateral renal ischemia for 45 min followed by reperfusion for 24 h. Group 1, Sham; group 2, I/R; group 3, L-arginine; and group 4, L-arginine+zinc protoporphyrin (ZnPP). The levels of serum creatinine (Scr), blood urea nitrogen (BUN), serum nitric oxide (NO), histic malondialdehyde (MDA) and reactive oxygen species (ROS) and superoxide dismutase (SOD) activity were determined, and the expression levels of Nrf2, HO-1, NF-κB, and HSP70 were evaluated. Results: The treatment of rats with L-arginine produced a significant reduction in the levels of BUN, Scr, MDA and a significant enhancement in the level of NO and in the activity of SOD compared to renal I/R groups. The expression levels of Nrf2, HO-1, and HSP70 were strongly increased, and the expression of NF-κB and production of ROS were significantly decreased in the L-arginine group compared to that of the I/R group. ZnPP increased renal damage and displayed effects opposite to those of L-arginine. Conclusion: These findings suggested that L-arginine/NO reduces renal dysfunction associated with I/R of the kidney and may act as a trigger to regulate the NF-κB, HSP70 and Nrf2/HO-1 signaling cascades.

Banhasasim-Tang Treatment Reduces the Severity of Esophageal Mucosal Ulcer on Chronic Acid Reflux Esophagitis in Rats.

The present study was conducted to evaluate both antioxidant and anti-inflammatory activity of Banhasasim-tang (BHSST) on chronic acid reflux esophagitis (CRE) model. Rat CRE model was established operatively and then treated with BHSST (1 g/kg body weight per day) for 15 days Esophageal pathological changes were analyzed using macroscopic examination and hematoxylin/eosin staining. The antioxidant and inflammatory protein levels were determined using Western blotting. The administration of BHSST significantly reduced both the overexpression of serum reactive oxygen species (ROS) and an excessive formation of thiobarbituric acid-reactive substances (TBARS) in esophagus tissue. Thus, the severity of esophageal ulcer was lower in BHSST treated rats than control rats on the gross and histological evaluation. Nuclear factor-erythroid 2-related factor 2 (Nrf2) led to the upregulation of antioxidant enzyme including SOD, GPx-1/2, and HO-1 by binding to antioxidant response element (ARE). Moreover, BHSST administration markedly reduced the expression of inflammatory proteins through mitogen-activated protein kinase- (MAPK-) related signaling pathways and decreased significantly the protein expressions of inflammatory mediators and cytokines by inhibition of nuclear factor-kappa B (NF-κB) activation. Taken together, these results support the fact that BHSST administration can suppress the development of esophageal mucosal ulcer via regulating inflammation through the activation of the antioxidant pathway.

Long-chain bases from sea cucumber mitigate endoplasmic reticulum stress and inflammation in obesity mice

Endoplasmic reticulum (ER) stress and inflammation can induce hyperglycemia. Long-chain bases (LCBs) from sea cucumber exhibit antihyperglycemic activities. However, their effects on ER stress and inflammation are unknown. We investigated the effects of LCBs on ER stress and inflammatory response in high-fat, fructose diet-induced obesity mice. Reactive oxygen species and free fatty acids were measured. Inflammatory cytokines in serum and their mRNA expressions in epididymal adipose tissues were investigated. Hepatic ER stress-related key genes were detected. c-Jun NH2-terminal kinase and nuclear factor κB inflammatory pathways were also evaluated in the liver. Results showed that LCBs reduced serum and hepatic reactive oxygen species and free fatty acids concentrations. LCBs decreased serum proinflammatory cytokines levels, namely interleukin (IL)-1β, tumor necrosis factor-α, IL-6, macrophage inflammatory protein 1, and c-reactive protein, and increased anti-inflammatory cytokine IL-10 concentration. The mRNA and protein expressions of these cytokines in epididymal adipose tissues were regulated by LCBs as similar to their circulatory contents. LCBs inhibited phosphorylated c-Jun NH2-terminal kinase and inhibitor κ kinase β, and nuclear factor κB nuclear translocation. LCBs also inhibited mRNA expression of ER stress markers glucose regulated protein, activating transcription factor 6, double-stranded RNA-activated protein kinase-like endoplasmic reticulum kinase, and X-box binding protein 1, and phosphorylation of eukaryotic initiation factor-α and inositol requiring enzyme 1α. These results indicate that LCBs can alleviate ER stress and inflammatory response. Nutritional supplementation with LCBs may offer an adjunctive therapy for RE stress-associated inflammation.

Oxidative stress drives CD8+ T-cell skin trafficking in patients with vitiligo through CXCL16 upregulation by activating the unfolded protein response in keratinocytes

In patients with vitiligo, an increased reactive oxygen species (ROS) level has been proved to be a key player during disease initiation and progression in melanocytes. Nevertheless, little is known about the effects of ROS on other cells involved in the aberrant microenvironment, such as keratinocytes and the following immune events. CXCL16 is constitutively expressed in keratinocytes and was recently found to mediate homing of CD8+ T cells in human skin. We sought to explicate the effect of oxidative stress on human keratinocytes and its capacity to drive CD8+ T-cell trafficking through CXCL16 regulation. We first detected putative T-cell skin-homing chemokines and ROS in serum and lesions of patients with vitiligo. The production of candidate chemokines was detected by using quantitative real-time PCR and ELISA in keratinocytes exposed to H2O2. Furthermore, the involved mediators were analyzed by using quantitative real-time PCR, Western blotting, ELISA, and immunofluorescence. Next, we tested the chemotactic migration of CD8+ T cells from patients with vitiligo mediated by the CXCL16-CXCR6 pair using the transwell assay. CXCL16 expression increased and showed a positive correlation with oxidative stress levels in serum and lesions of patients with vitiligo. The H2O2-induced CXCL16 expression was due to the activation of 2 unfolded protein response pathways: kinase RNA (PKR)-like ER kinase-eukaryotic initiation factor 2α and inositol-requiring enzyme 1α-X-box binding protein 1. CXCL16 produced by stressed keratinocytes induced migration of CXCR6+CD8+ T cells derived from patients with vitiligo. CXCR6+CD8+ T-cell skin infiltration is accompanied by melanocyte loss in lesions of patients with vitiligo. Our study demonstrated that CXCL16-CXCR6 mediates CD8+ T-cell skin trafficking under oxidative stress in patients with vitiligo. The CXCL16 expression in human keratinocytes induced by ROS is, at least in part, caused by unfolded protein response activation.

Forsythia suspensa extract attenuates lipopolysaccharide-induced inflammatory liver injury in rats via promoting antioxidant defense mechanisms.

Reactive oxygen species (ROS) have been shown to have a role in inflammation. We investigated whether Forsythia suspensa extract (FSE) could exert its antioxidant potential against lipopolysaccharide (LPS)-induced inflammatory liver injury in rats. Rats were orally fed FSE once daily for 7 consecutive days prior to LPS (Escherichia coli, serotype O55:B5) injection. LPS treatment caused liver dysfunction as evidenced by massive histopathological changes and increased serum alanine aminotransferase and aspartate aminotransferase activities which were ameliorated by FSE pretreatment. FSE attenuated LPS-induced depletion of cytosolic nuclear factor-erythroid 2-related factor 2 (Nrf2) and suppression of Nrf2 nuclear translocation in liver, and the generation of ROS and malondialdehyde in serum and liver. FSE increased the Nrf2-mediated induction of heme oxygenase-1 in liver, as well as superoxide dismutase and glutathione peroxidase activities in serum and liver. Importantly, FSE attenuated LPS-induced nuclear factor-кB (NF-кB) nuclear translocation in liver, and subsequently decreased tumor necrosis factor-α, interleukin (IL)-1β and IL-6 levels in serum and liver, which were associated with FSE-induced activation of Nrf2 in liver. These results indicate that the protective mechanisms of FSE may be involved in the attenuation of oxidative stress and the inhibition of the NF-кB-mediated inflammatory response by modulating the Nrf2-mediated antioxidant response against LPS-induced inflammatory liver injury.

흑삼 열수 추출물의 항산화 효과를 통한 염산/에탄올로 유발된 위염 억제 작용

Black ginseng (BG) obtained by a 9-fold steaming process of Panax ginseng has been reported to have anti-oxidative, anti-obesity, and anti-diabetes effects. The current study evaluated the protective effect of BG by steaming time in an HCl/ethanol-induced acute gastritis model. BG was divided into four samples according to steaming-drying processing (Gin1, Gin3, Gin6, and BG). High performance liquid chromatography analysis, free radical scavenging activity, and total phenol and flavonoid contents were examined in ginseng and four BG samples. Compared with ginseng, BG showed a stronger radical scavenging effect and higher contents of total phenol and flavonoids. To evaluate the anti-gastritic effect of BG, mice were distributed into five groups: normal mice (N), acute gastritic mice with distilled water (CON), acute gastritic mice with 100 mg/kg of ginseng (Gin0), acute gastritic mice with 100 mg/kg of BG (BG), and acute gastritic mice with 10 mg/kg of sucralfate (SC). After 1 hour of pre-treatment with water, extracts (Gin0 and BG), or drug (SC), experimental groups except for N were orally administered 0.5 mL of 150 mM HCl/60% ethanol (v/v) mixture. Blood was collected 1 hour later from the heart, and gastric tissue was harvested. Reactive oxygen species (ROS) levels were measured in serum, and related protein expression was examined by Western blot assay. In HCl/ethanol-induced acute gastritic mice, treatment with ginseng or BG improved mucosal damage in the histological evaluation. The serum ROS level significantly decreased in the BG-treated group compared with the CON group. Furthermore, expression of inflammatory cytokines significantly decreased in the BG-treated group compared with the CON group. Based on these results, antioxidant and anti-gastritic activities of ginseng were enhanced by streaming-drying processing, in part due to an increase in biological active compounds.

Effects of oregano essential oil or quercetin supplementation on body weight loss, carcass characteristics, meat quality and antioxidant status in finishing pigs under transport stress

This study compared the effects of dietary oregano essential oil (OEO), quercetin or vitamin E (vit E), on the live body weight loss, carcass characteristics, meat quality and antioxidant status of pigs after transportation. A total of 340 finishing pigs (Large White×Landrace) with an initial body weight of 74kg (±4.0kg) were randomly assigned to one of four treatment groups (five replicate pens per treatment, 17 pigs per pen). Pigs consumed the basal diet (control) or the basal diet supplemented with 200mg vit E/kg (positive control), 25mg OEO/kg or 25mg quercetin/kg for 4 weeks. After this period, 144 pigs (36 pigs per treatment) were transported for 5h before slaughter. Compared with the control group, the OEO or quercetin groups had a higher average daily gain (P<0.05), and the OEO group also had a lower feed intake/gain (P<0.05). The live body weight loss was less in the OEO group after 5h transportation (P<0.05) than in the control group. The hot carcass weight and dressing percentage were higher in the OEO group after 5h of transportation (P<0.05) than in the control group. After slaughter, the pH value at 45-min postmortem and Opto-star value (meat color) at 24-h postmortem increased in the vit E, OEO or quercetin groups (P<0.05) compared with the control group. The vit E or quercetin groups also exhibited higher 24-h postmortem pH values (P<0.05) than the control group. The Longissimus thoracis et lumborum muscle of pigs from the OEO or quercetin groups produced lower 24-h drip loss values (P<0.05) than that of pigs from the control group. Compared with the control group, the OEO or quercetin groups had reduced levels of TBARS (thiobarbituric acid reactive substances) and ROS (reactive oxygen species) in serum, muscle and liver (P<0.05), while the vit E group had reduced levels in serum only (P<0.05). The OEO or quercetin groups also had increased levels of Gpx (glutathione peroxidase) and T-SOD (total superoxide dismutase) activity in serum and liver compared with the control group (P<0.05). Conversely, there were no differences between the vit E and control groups in Gpx or T-SOD activities. In conclusion, supplementation with dietary OEO or quercetin may be superior to supplementation with dietary vit E in alleviating the negative effects of transportation on pigs by improving the pigs’ antioxidant status.

The effects of aerobic exercise training on oxidant–antioxidant balance, neurotrophic factor levels, and blood–brain barrier function in obese and non-obese men

PurposeThe purpose of this study was to investigate the effects of obesity and aerobic exercise training on oxidant–antioxidant balance, neurotrophic factor levels, and blood–brain barrier (BBB) function. MethodsTen non-obese healthy men (body mass index < 25 kg/m2) and 10 obese men (body mass index ≥ 25 kg/m2) were included in the study. Both groups performed treadmill exercise for 40 min 3 times weekly for 8 weeks at 70% heart rate reserve. Blood samples were collected to examine oxidant–antioxidant balance (reactive oxygen species (ROS) and superoxide dismutase (SOD) activity levels), neurotrophic factors (brain-derived neurotrophic factor (BDNF), nerve growth factor, and glial cell line-derived neurotrophic factor levels), and BBB function (S100β and neuron-specific enolase (NSE) levels) before and after exercise training. ResultsThe obese group showed significantly greater changes than the non-obese group in serum ROS (−0.46 ± 0.31 mmol/L vs. −0.10 ± 0.17 mmol/L, p = 0.005), serum S100β levels (−8.50 ± 5.92 ng/L vs. −0.78 ± 5.45 ng/L, p = 0.007), and serum NSE levels (−0.89 ± 0.54 µg/L vs. −0.01 ± 0.74 µg/L, p = 0.007) after training. At baseline, the obese group showed significantly higher serum ROS and S100β levels and significantly lower serum SOD activity and BDNF levels than the non-obese group (p < 0.05). The obese group showed significantly lower serum ROS, S100β, and NSE levels and significantly higher serum SOD activity and BDNF levels after training compared with baseline (p < 0.05). ConclusionThese results suggest that obesity can reduce serum neurotrophic factor levels and can induce BBB dysfunction. On the other hand, aerobic exercise can improve an oxidant–antioxidant imbalance in obese subjects and limit BBB dysfunction.

Obesity promotes oxidative stress and exacerbates blood-brain barrier disruption after high-intensity exercise

PurposeThe purpose of this study was to investigate the effects of obesity and high-intensity acute exercise on oxidant-antioxidant status, neurotrophic factor expression, and blood-brain barrier (BBB) disruption. MethodsTwenty-four healthy, untrained men (12 non-obese (mean 14.9% body fat) and 12 obese subjects (mean 29.8% body fat)) performed 20 min of continuous submaximal aerobic exercise at 85% maximal oxygen consumption. Blood sampling was performed to examine the oxidant-antioxidant status (reactive oxygen species (ROS) and superoxide dismutase (SOD)), neurotrophic factors (brain-derived neurotrophic factor (BDNF) and nerve growth factor (NGF)), and BBB disruption (S100β and neuron-specific enolase) before and after acute exercise. ResultsThe obese group showed significantly higher pre-exercise serum ROS levels and significantly lower pre-exercise serum SOD levels than the non-obese group (p < 0.05). Serum ROS, SOD, BDNF, NGF, and S100β levels were significantly increased post-exercise compared with pre-exercise levels in both the non-obese and the obese groups (p < 0.05). The obese group showed significantly higher serum ROS, BDNF, NGF, and S100β levels post-exercise compared to the non-obese group (p < 0.05). ConclusionOur study suggests that episodic vigorous exercise can increase oxidative stress and blood neurotrophic factor levels and induce disruption of the BBB. Moreover, high levels of neurotrophic factor in the blood after exercise in the obese group may be due to BBB disruption, and it is assumed that oxidative stress was the main cause of this BBB disruption.

Hyperuricemia Is an Independent Risk Factor for Erectile Dysfunction

IntroductionHyperuricemia may be related to the development of endothelial dysfunction and cardiovascular diseases. However, the association between hyperuricemia and erectile dysfunction (ED) is not currently clear. AimThe goal of this study is to investigate the effect of hyperuricemia on erectile function and possible mechanisms. MethodsTwenty-four 8-week-old male SD rats were randomly divided into 4 groups. Group A (control): Rats received normal saline and served as controls. Group B (hyperuricemia): rats were given oxonic acid 250 mg/kg bw/day through gastric gavage for 4 weeks. Group C (febuxostat): normal rats were treated with 5 mg/kg febuxostat through gastric gavage for 4 weeks. Group D (hyperuricemia + Febuxostat): normal rats were treated with 250 mg/kg bw/day oxonic acid and 5 mg/kg bw/day febuxostat with 1 hour interval for 4 weeks. MeasurementsThe level of serum uric acid, the maximum intracavernosal pressure (ICPmax), mean arterial pressure (MAP), and the expression of endothelial nitric oxide synthase (eNOS), phospho-eNOS, neuronal NOS, Rho-associated protein kinaise (ROCK)1 and ROCK2 and the level of nitric oxide (NO) and reactive oxygen species (ROS) in cavernous tissue were determined. ResultsThe level of serum uric acid and ROS in hyperuricemic rats was significantly higher than that in the other 3 groups (P < .05). After electrostimulation with 3 and 5 voltage, the ratio of ICPmax/MAP in hyperuricemic rats was significantly less than that in other 3 groups (P < .05), respectively. eNOS, p-eNOS, and nNOS expression in hyperuricemic rats were significantly decreased compared to the other 3 groups (P < .05), respectively. ConclusionErectile function is impaired by hyperuricemia. The decrease of eNOS, p-eNOS, and nNOS protein expression and increase of ROS in cavernous tissue may be one of the key mechanisms of ED caused by hyperuricemia.

DL-3-n-butylphthalide delays the onset and progression of diabetic cataract by inhibiting oxidative stress in rat diabetic model.

DL-3-n-butylphthalide (NBP) is a therapeutic drug used for ischemic stroke treatment. Here, we investigated the impact of NBP on the development of rat diabetic cataract induced by intraperitoneal injection of streptozotocin (STZ). NBP was then administrated by oral gavage for nine weeks. Cataract development was monitored through ophthalmoscope inspections. The levels of blood glucose and serum reactive oxygen species (ROS), malondialdehyde (MDA) and 8-Hydroxydeovexyguanosine (8-OHdG) were measured. Total and soluble protein and oxidative stress parameters, such as 2, 4- dinitrophenylhydrazone (DNP), 4-hydroxynonenal (4-HNE) and MDA in the lenses were determined by Western blot and thiobarbituric acid analyses. The expressions of NF-E2-related factor 2 (Nrf2) and its downstream antioxidant enzymes, thioredoxin (TRX), Catalase and nuclear accumulation of Nrf2 were determined by Western blot and immunohistochemistry analyses. We showed that NBP treatment significantly improved the cataract scores, the levels of DNP, 4-HNE, and MDA in the lens compared to the non-treated groups. NBP also enhanced the expressions of Nrf2, TRX and catalase in the lens of diabetic rats. In addition, NBP treatment also decreased levels of blood glucose, serum MDA and 8-OHdG. These results suggested that NBP treatment significantly delayed the onset and progression of diabetic cataract by inhibiting the oxidative stresses.

Probucol Protects Endothelial Progenitor Cells Against Oxidized Low-Density Lipoprotein via Suppression of Reactive Oxygen Species Formation In Vivo

Background/Aims: Oxidized low-density lipoprotein (ox-LDL) is a major component of hyperlipidemia and contributes to atherosclerosis. Endothelial progenitor cells (EPCs) play an important role in preventing atherosclerosis and notably decreased in hyperlipidemia. Ox-LDL and ox-LDL-related reactive oxygen species (ROS) have deleterious effects on EPCs. Probucol as an antioxidant and anti-inflammatory drug reduces ROS production. The present study was to determine if probucol could protect EPCs from ox-LDL in vivo and to investigate the potential mechanisms. Methods: ox-LDL was injected into male C57BL/6 mice for 3 days with or without probucol treatment with PBS as control. Bone marrow (BM) fluid, serum, circulating mononuclear cells (MNCs) and EPCs were collected for analysis. Results: the increased extracellular ROS in BM, serum and blood intracellular ROS production in the mice with ox-LDL treatment in association with a significant reduction of circulating MNCs and EPCs were restored with Probucol treatment. A significant increase in the serum ox-LDL and C-reactive protein and decrease in superoxide dismutase and circulating MNCs and EPCs were observed in hyperlipidemic patients that were effectively reversed with probucol treatment. Conclusion: these data suggested that probucol could protect EPCs from ox-LDL through inhibition of ROS production in vivo.

Kangen-karyu raises surface body temperature through oxidative stress modification.

Kangen-karyu, a prescription containing six herbs, has been shown to achieve its pharmacological effect through oxidative stress-dependent pathways in animal models. The aim of this study is to investigate the relationship between the antioxidative effect and pharmacological mechanisms of Kangen-karyu, specifically its body temperature elevating effect in humans. Healthy human volunteers, age 35 ± 15 years old, were enrolled in this study. Surface body temperature, serum nitrite, reactive oxygen species (ROS) scavenging activities, and inflammatory cytokines were investigated before and 120 min after Kangen-karyu oral intake. Kangen-karyu significantly increased the surface-body temperature of the entire body; this effect was more remarkable in the upper body and continued for more than 120 min. Accompanying this therapeutic effect, serum nitrite levels were increased 120 min after oral administration. Serum ROS scavenging activities were enhanced against singlet oxygen and were concomitantly decreased against the alkoxyl radical. Serum nitrite levels and superoxide scavenging activities were positively correlated, suggesting that Kangen-karyu affects the O2•−-NO balance in vivo. Kangen-karyu had no effect on IL-6, TNF-α and adiponectin levels. These results indicate that the therapeutic effect of Kangen-karyu is achieved through NO- and ROS-dependent mechanisms. Further, this mechanism is not limited to ROS production, but includes ROS-ROS or ROS-NO interactions.

Age-dependency of the serum oxidative level in the senescence-accelerated mouse prone 8.

Systemic oxidative stress is considered to cause aging. In this study, to estimate the oxidative stress level in senescence-accelerated mouse prone 8 (SAMP8), we evaluated serum reactive oxygen species production and reduction capacity by measurement of Diacron-Reactive Oxygen Metabolites (d-ROM) and Biological Antioxidant Potential (BAP), respectively, with age. SAMP8 showed earlier increase of d-ROM value with age than SAM resistant 1 (SAMR1), the control strain. The BAP level was the highest in adult SAMP8, whereas SAMR1 presented the sustained BAP values between ages. These results indicate that oxidative stress in SAMP8 is higher than SAMR1. Our study is the first detailed report about d-ROM and BAP in SAMP8 and will provide useful fundamental data for future aging studies.

Antioxidant and Anti-Inflammatory Effects of Rhei Rhizoma and Coptidis Rhizoma Mixture on Reflux Esophagitis in Rats.

The purpose of this study was to investigate the antioxidant and anti-inflammatory effects of the combined extract of Rhei rhizoma and Coptidis rhizoma (RC-mix) in experimental model of acute reflux esophagitis. The antioxidant activity was assessed by in vitro 2,2-diphenyl-1-picrylhydrazyl (DPPH) and 2,2′-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) assays. RC-mix was given at 100, 200, and 400 mg/kg body weight 2 h prior to induction of reflux esophagitis (RE). After 5 h, the effects of RC-mix treated rats were compared with those of normal and control rats. The representative flavonoid contents of RC-mix, such as sennoside A, epiberberine, coptisine, palmatine, and berberine, were detected using HPLC. The elevated esophageal mucosa damage was markedly ameliorated by RC-mix treatment in a dose-dependent manner. Furthermore, the administration of RC-mix reduced the increase of serum reactive oxygen species (ROS) and peroxynitrite (ONOO−). The improvement of superoxide dismutase (SOD) and heme oxygenase-1 (HO-1) levels were marked in the group given RC-mix. Moreover, the elevation of inflammatory mediators and cytokines by nuclear factor-kappa B (NF-κB) activation in control rats decreased by RC-mix pretreatment. These results indicate that RC-mix treatment reduces the pathological states of esophagitis via regulating NF-κB mediated inflammation related to oxidative stress.

Inhibition of complement C3 might rescue vascular hyporeactivity in a conscious hemorrhagic shock rat model

BackgroundVascular hyporeactivity in severe hemorrhagic shock could induce refractory hypotension and is an important cause of death. The global acute inflammatory response induced in shock triggers the over-expression of reactive oxygen species, NO, ET1 and TNF-α, which play essential roles in the pathology of vascular hyporeactivity. This leads to a hypothesis that inhibition of the complement system, the mediator of the inflammatory cascade, might be a promising therapeutic exploration for vascular hyporeactivity. MethodsWe use cobra venom factor (CVF) and the soluble form of CR1 (sCR1) which deplete or inhibit complement C3 respectively to examine its role in vascular hyporeactivity in a conscious hemorrhagic shock rat model. ResultsWe first confirmed the over-activation of C3 during shock and the down-regulation effects of CVF and sCR1 on C3. Then, both CVF and sCR1 could significantly mitigate the over-expression of serum NO, ET-1, TNF-α and reactive oxygen species. Finally, the vascular reactivity of superior mesenteric arteries (SMA) was examined in vitro, which confirmed the massive reduction of vascular reactivity in shock, which was significantly rescued by both CVF and sCR1. ConclusionsInhibition of C3 might improve the reactivity of SMA to norepinephrine during hemorrhagic shock possibly through the downregulation of NO, ET1, TNF-α and reactive oxygen radicals.

The impact of detoxifying and repair gene polymorphisms and the levels of serum ROS in the susceptibility to multiple sclerosis

ObjectiveMultiple sclerosis (MS) is an autoimmune neurodegenerative disease with unknown etiology. Oxidative stress (OS) has been implicated to play a role in its cause; therefore, antioxidants and repair systems may help in restoring oxidant–antioxidant balance. Since polymorphisms in DNA repair genes can result in reduced DNA repair capacity, it is important to investigate its association with OS products to demonstrate the impact of individual susceptibility. Our aim is to examine whether a defect in one of the detoxifying and DNA repair enzyme systems could explain the association between MS and exposure to OS products. MethodsWe investigated the association of polymorphisms in the metabolizing and DNA repair genes with serum Reactive Oxygen Species (ROS) levels. Gene polymorphisms were analyzed by simultaneous multiplex and Restriction Fragment Length Polymorphism Polymerase Chain Reaction and serum ROS levels were detected. ResultsOGG1 Ser/Cys and Ser/Cys+Cys/Cys genotypes had higher MS risk. XRCC1 Arg/Gln+Gln/Gln genotype increased the risk of MS. ConclusionsOur data suggested that OGG1 Ser326Cys gene polymorphism is a major genetic factor involved in the development of MS. Smoking is also a pivotal confounding factor for subjects with mutant genotypes of XRCC1 Arg399Gln. Further studies are needed to reaffirm our results.

Oxidative status in treatment-naïve essential thrombocythemia: a pilot study in a single center

Oxidative stress (OS), due to pro-oxidant species [reactive oxygen species (ROS)] excess not counterbalanced by endogenous antioxidant molecules [e.g., reduced glutathione (GSH)], is involved in the pathogenesis of human cancers, but few data are available on essential thrombocythemia (ET). This study aims to investigate OS in ET off-therapy patients. Thirty ET treatment-naïve patients were compared with 26 age-matched and gender-matched controls. Serum ROS, urinary 8-hydroxydeoxyguanosine, full blood GSH levels, and reduced/oxidized GSH ratio (GSH/GSSG) were measured. Data were adjusted for gender, age, JAK2 mutational status, smoking, dyslipidemia, or hypercholesterolemia requiring drug therapy, antiplatelet therapy, treatment with acetylsalicylic acid, high-sensitive C-reactive protein levels, and absolute monocyte count. ROS and GSH levels were increased in both patients and controls. Patients showed increased GSSG (p = 0.05), reduced GSH/GSSG ratio (p = 0.08), and similar 8-hydroxydeoxyguanosine levels when compared with controls. No differences in OS parameters were found between JAK2-positive and JAK2-negative patients. Confounding factors did not modify the results. Our study suggests an OS condition in a cohort of treatment-naïve ET patients, not associated with JAK2 mutational status or with chronic inflammation situation. GSH/GSSG ratio, altered in ET patients because of increased GSSG levels, showed the presence of higher GSH levels in ET than controls as a possible compensatory mechanism of an excess of pro-oxidant production. Copyright © 2015 John Wiley & Sons, Ltd.