Th17 response to Dermatophagoides pteronyssinus is related to late-phase airway and systemic inflammation in allergic asthma

BackgroundTh17 cells may play a role in the development of late-phase allergen-induced airway and systemic inflammation in allergic asthma, although the mechanisms involved remain to be elucidated.MethodsA total of 36 subjects were enrolled into the study: 15 allergic asthma patients with early asthmatic reaction (n = 7) or dual asthmatic reaction (n = 8) developed to inhaled D. pteronyssinus, 13 patients with allergic rhinitis, and 8 healthy subjects. Peripheral blood and induced sputum were collected 24 h before as well as 7 h and 24 h after a bronchial challenge with D. pteronyssinus. Th17 cells were analyzed by FACS; IL-17 levels were determined by ELISA.ResultsAt baseline, the percentage of peripheral blood Th17 cells and serum and sputum IL-17 levels were significantly higher in all groups of studied patients compared with those of healthy subjects. After the bronchial challenge, there was a significant increase in the percentage of peripheral blood Th17 cells and in serum and sputum IL-17 levels in rhinitis and asthma patients compared with their baseline values, particularly in allergic asthma patients with the dual asthmatic reaction. Positive correlations were found between the percentage of Th17 cells and IL-17 levels in serum (Rs = 0.649; P = 0.009) as well in sputum (Rs = 0.583; P = 0.022) in allergic asthma patients 24 h after the bronchial challenge.ConclusionsThe Th17 response is associated with the development of late-phase airway and systemic inflammation after the inhalation of D. pteronyssinus in patients with allergic asthma.     

Mesenchymal stem cells upregulate Treg cells via sHLA-G in SLE patients

BackgroundSoluble human leukocyte antigen-G (sHLA-G) is a non-classical HLA class I molecule, exhibiting strong immunosuppressive properties by inducing the differentiation of T regulatory cells (Treg). Mesenchymal stem cells (MSCs) transplantation alleviates disease progression in systemic lupus erythematosus (SLE) patients. However, the underlying mechanisms are largely unknown.ObjectivesTo explore whether sHLA-G is involved in upregulating effects of MSCs on Treg, which contributes to therapeutic effects of MSCs transplantation in SLE.MethodsThe serum sHLA-G levels of SLE patients and healthy controls were detected by ELISA. The percentages of peripheral blood CD4 + ILT2 +, CD8 + ILT2 +, CD19 + ILT2 + cells and Treg cells were examined by flow cytometry. Ten patients with active SLE, refractory to conventional therapies, were infused with umbilical cord derived MSCs (UC-MSCs) and serum sHLA-G was measured 24 h and 1 month after infusion. The mice were divided into three groups: C57BL/6 mice, B6.MRL-Fas^lpr mice infused with phosphate buffer saline (PBS), and B6.MRL-Fas^lpr mice infused with bone marrow MSCs (BM-MSCs). Then, the concentrations of serum Qa-2 were detected. Peripheral blood mononuclear cells (PBMCs) were isolated from SLE patients and co-cultured with UC-MSCs for 3 days at different ratios (50:1, 10:1, and 2:1) with or without HLA-G antibody, and the frequencies of CD4 + CD25 + Foxp3 + T cells were then determined by flow cytometry.ResultsThe concentrations of serum sHLA-G were comparable between SLE patients and healthy controls. However, there was a negative correlation between sHLA-G levels and SLE disease activity index (SLEDAI) scores in active SLE patients (SLEDAI > 4). We found that serum sHLA-G levels were negatively correlated with blood urea nitrogen, serum creatinine and 24-hour urine protein in SLE patients. The sHLA-G levels were significantly lower in SLE patients with renal involvement than those without renal involvement. The expression of ILT2 on CD4 + T cells from SLE patients decreased significantly compared to that of healthy controls. A positive correlation between the frequencies of Treg and CD4 + ILT2 + T cells was found in SLE patients. The levels of sHLA-G increased 24 h post UC-MSCs transplantation. The concentrations of Qa-2 in BM-MSCs transplanted mice were significantly higher than those of control group. In vitro studies showed that MSCs increased the frequency of Treg cells in SLE patients in a dose-dependent manner, which was partly abrogated by the anti-HLA-G antibody.ConclusionsOur results suggested that MSCs may alleviate SLE through upregulating Treg cells, which was partly dependent on sHLA-G.     

The effect of Phloretin on human γδ T cells killing colon cancer SW-1116 cells

ObjectiveTo explore the effect and mechanism of Phloretin on human γδ T cells killing colon cancer SW-1116 cells.Methodsγδ T cells were amplified in vitro from human peripheral blood mononuclear cells through isopentenyl pyrophosphate method (IPP). After cocultured different concentrations of Phloretin with γδ T cells or SW-1116 cells for 48 h respectively, MTT assay was used to test the growth curve of these two cells; Flow cytometry to test the expression of Granzyme B (GraB), perforin (PFP), CD107a and IFN-γ of γδ T cells; Lactate dehydrogenase (LDH) release assay to test the cytotoxic activity of the γδ T cells on SW-1116 cells; and Western blot to test the Wnt3a expression of the γδ T cells.ResultsAfter cultured with IPP for ten days, the percentage of γδ T cells increased from 3.31 ± 3.00% to 78.40 ± 10.30%. Compared to the control group, when the concentration of Phloretin increased from 2.35 μg/ml to 18.75 μg/ml, it could significantly proliferate the γδ T cell growth (P < 0.05) and inhibit the growth of SW-1116 cells in dose–response, and the expression of GraB, PFP, CD107a and Wnt3a significantly increased (P < 0.05). Significant positive relationships were observed among CD107a and PFP, GraB, cytotoxicity (P < 0.05). The percentage of IFN-γ producing γδ T cells treated with Phloretin was significantly higher than control group.ConclusionPhloretin can enhance the killing effect of γδ T cells on SW-1116 cells; the mechanism may be that Phloretin could proliferate the γδ T cell growth, increase the expression of PFP and GraB, activate the Wnt signaling pathway, and produce higher level of IFN-γ. Indeed CD107a expression probably correlates quite well with antitumor activity.     

8-Isoprostane levels in serum and bronchoalveolar lavage in idiopathic pulmonary fibrosis and sarcoidosis

Objective and designSarcoidosis and idiopathic pulmonary fibrosis (IPF) are both associated with deregulated inflammatory mechanisms partially triggered by aggravated oxidative stress. 8-Isoprostane has been proposed as a reliable marker of oxidative stress, linked to several pulmonary diseases. We aimed to explore differences in 8-isoprostane levels in IPF and sarcoidosis patients, and controls.MethodsWe included 16 IPF and 55 sarcoidosis patients, as well as 17 controls in the study. 8-Isoprostane levels were measured in serum and in bronchoalveolar lavage (BAL).ResultsSerum 8-isoprostane levels were increased in all patient groups vs controls (p < 0.001). The systemic 8-isoprostane concentrations were higher in sarcoidosis patients as compared to IPF subjects and controls (p = 0.017 and p < 0.001, respectively). IPF patients exhibited increased serum 8-isoprostane levels when compared to controls (p < 0.001). Sarcoidosis patients presented significantly increased 8-isoprostane BAL levels when compared to IPF patients (p = 0.002).ConclusionOur data indicate that the level of oxidative stress, as reflected by 8-isoprostane concentrations, is enhanced in patients with sarcoidosis, and to a lesser extent, in IPF patients when compared to controls, suggesting a potential implication of redox imbalance in both diseases.     

Bisphenol A exposure increases liver fat in juvenile fructose-fed Fischer 344 rats

BackgroundPrenatal exposure to bisphenol A (BPA) has been shown to induce obesity in rodents. To evaluate if exposure also later in life could induce obesity or liver damage we investigated these hypothesises in an experimental rat model.MethodsFrom five to fifteen weeks of age, female Fischer 344 rats were exposed to BPA via drinking water (0.025, 0.25 or 2.5 mg BPA/L) containing 5% fructose. Two control groups were given either water or 5% fructose solution. Individual weight of the rats was determined once a week. At termination magnetic resonance imaging was used to assess adipose tissue amount and distribution, and liver fat content. After sacrifice the left perirenal fat pad and the liver were dissected and weighed. Apolipoprotein A-I in plasma was analyzed by western blot.ResultsNo significant effects on body weight or the weight of the dissected fad pad were seen in rats exposed to BPA, and MRI showed no differences in total or visceral adipose tissue volumes between the groups. However, MRI showed that liver fat content was significantly higher in BPA-exposed rats than in fructose controls (p = 0.04). BPA exposure also increased the apolipoprotein A-I levels in plasma (p < 0.0001).ConclusionWe found no evidence that BPA exposure affects fat mass in juvenile fructose-fed rats. However, the finding that BPA in combination with fructose induced fat infiltration in the liver at dosages close to the current tolerable daily intake (TDI) might be of concern given the widespread use of this compound in our environment.     

Effect of natural porcine surfactant in Staphylococcus aureus induced pro-inflammatory cytokines and reactive oxygen species generation in monocytes and neutrophils from human blood

Surfacen® is a clinical surfactant preparation of porcine origin. In the present study, we have evaluated the effect of Surfacen® in the modulation of oxidative burst in monocytes and neutrophils in human blood and pro-inflammatory cytokine production in peripheral blood mononuclear cells (PBMC). Reactive oxygen species (ROS) level was measured in monocytes and neutrophils by flow cytometry using 2,7-dichlorofluorescein diacetate (DCFH-DA) as substrate, while, tumor necrosis factor (TNF)-α and interleukin (IL)-6 levels were estimated in PBMC supernatant by enzyme-linked immunosorbent assays (ELISA). Our results show that Staphylococcus aureus-induced ROS level was slightly affected by Surfacen® added to whole blood monocytes and neutrophils. The time course experiments of pre-incubation with Surfacen® showed no significant increase of ROS level at 2 h; however, the ROS level decreased when pre incubated for 4 h and 6 h with Surfacen®. Pre-incubation of PBMC cells with Surfacen® at 0.125 and 0.5 mg/mL showed a dose-dependent suppression of TNF-α levels measured after 4 h of S. aureus stimulation, an effect less impressive when cells were stimulated for 24 h. A similar behavior was observed in IL-6 release. In summary, the present study provides experimental evidence supporting an anti-inflammatory role of Surfacen® in human monocytes and neutrophils in vitro.     

The comparative effects of perindopril and catechin on mesangial matrix and podocytes in the streptozotocin induced diabetic rats

BackgroundHyperglycemia and advanced glucose end substance (AGE) are responsible for excessive reactive oxygen species (ROS) production, which causes oxidative stress in diabetes mellitus. Oxidative stress and high blood pressure may cause injury and glomerulosclerosis in the kidney. End-stage kidney failure induced by glomerulosclerosis leads to microalbuminuria (Ma) in diabetic nephropathy. We investigated the effects of an angiotensin converting enzyme inhibitor (ACEI), perindopril, and an antioxidant, catechin, on podocytes and the glomerular mesangial matrix in experimental diabetic nephropathy using ultrastructural visualization and immunohistochemical staining.MethodsWe compared 5 groups of male adult Wistar albino rats: a control group, an untreated diabetic group, and diabetic groups treated with perindopril, catechin, or catechin + perindopril.ResultsBlood glucose values in all diabetic groups were significantly higher than in the control group (p < 0.001). The body weight in all diabetic groups was significantly lower than in the control group (p < 0.001, p < 0.05). The kidney weight in the catechin + perindopril-treated diabetic group was significantly lower than in the untreated diabetic group (p < 0.001). In all treated diabetic groups, Ma levels decreased significantly (p < 0.001). Mesangial matrix and podocyte damage increased in the untreated diabetic group, but the group treated with catechin + perindopril showed less damage. TGF-beta 1 immunostaining was significantly lower in the catechin-treated and perindopril-treated groups than in the untreated diabetic group (p < 0.001). Catechin was more effective than ACEI in preventing podocyte structure. Podocytes appeared to be the first cells affected in diabetes mellitus. When exposed to hyperglycemia, podocytes caused the mesangial matrix to expand.ConclusionsCatechin and perindopril were more effective in preventing renal corpuscle damage when administered together.     

Pre-treatment with melatonin decreases abamectin induced toxicity in a nocturnal insect Spodoptera litura (Lepidoptera: Noctuidae)

AimOxidative stress is an important component of the mechanism of pesticide toxicity. The aim of the present study was to investigate the time-dependent melatonin effects against abamectin-induced oxidative stress in a S.litura model. Larvae were divided into 5 different groups; (1) control group,(2) Melatonin group (4.3 × 10⁻⁵ M/100 ml diet), (3) Abamectin group 1.5 ml/L, (4) Pre-melatonin treated group (PM) (4.3 × 10⁻⁵ M/100 ml diet) before abamectin exposure 1.5 ml/L, (5) Post-melatonin treated group (TM) after abamectin exposure. Melatonin was supplemented via artificial diet in PM and TM animals during 24 h.Main methodsMidgut, fatbody, and hemolymph, were collected for the analysis of oxidative stress markers (Total ROS, GSH, nitrite, TBARS, LPO), antioxidant enzyme levels (SOD, GST, CAT, POX, APOX) in fifth instar larvae. Midgut damage was examined by using morphological analysis.Key findingsOur results observed that ABA group showed significant changes (p < 0.001) in the ROS and carbonyl content in midgut. The increase of antioxidant enzyme levels (SOD, CAT, POX, and APOX) in midgut was led by the continuous free radical scavenger cascade of melatonin. Significant (p < 0.01) increases in CAT and APOX levels were seen in the fatbody of PM and TM treated insects.SignificanceIn conclusion, the results of the study revealed that abamectin toxicity generates oxidative stress in the insect, while pre-melatonin treatment reduces this damage due to its antioxidant properties, especially POX levels in midgut, fatbody, and hemolymph. Therefore, indoleamine can play a vital role curtailing the abamectin toxicity in time dependent manner in S.litura.     

Influence of specific endothelin-1 receptor blockers on hemodynamic parameters and antioxidant status of plasma in LPS-induced endotoxemia

BackgroundThe potent vasoconstrictor endothelin-1 has been implicated in the pathogenesis of plasma oxidative stress seen in sepsis. The selective endothelin receptor blockers BQ123 andBQ788 were used to investigate the importance of selective endothelin receptor blockage in modulating oxidative stress during endotoxemia.MethodsThe study was performed on male Wistar rats (n = 6 per group) divided into groups: (1) saline, (2) lipopolysaccharide (LPS) (15 mg\kg)-saline, (3) BQ123 (0.5 mg\kg)-LPS, (4) BQ123 (1 mg\kg)-LPS, (5) BQ788 (3 mg\kg)-LPS. The endothelin receptor type A (ETA-R) or type B (ETB-R) antagonist was injected intravenously 30 min before LPS administration. Blood pressure was monitored and blood was taken before, 90 min and 300 min after saline or LPS administration.ResultsInjection of LPS alone resulted in a decrease in mean arterial pressure (MAP) (p < 0.05), a decrease in ferric reducing ability of plasma (FRAP) value (p < 0.01) and a marked increase in plasma tumor necrosis factor α (TNF-α) and thiobarbituric acid reactive substances (TBARS) (p < 0.001, p < 0.001, respectively). Administration of BQ123 before LPS administration deteriorated MAP in a dose dependent way. Moreover, BQ123 (1 mg\kg) decreased plasma level of TBARS and TNF-α (p < 0.01 and p < 0.05, respectively) and increased FRAP value (p < 0.001). On the contrary, BQ788 prevented LPS-induced decrease in MAP (p < 0.001) and led to a significant reduction in plasma TBARS concentration (p < 0.01).ConclusionOur study showed that blockage of ETB-R during endotoxemia improved blood hemodynamics and decreased plasma lipid peroxidation. Blockage of ETA-R improved plasma antioxidant status and decreased lipid peroxidation and TNF-a production, but it deteriorated hemodynamic conditions.     

Protective effects of selenium on oxidative damage and oxidative stress related gene expression in rat liver under chronic poisoning of arsenic

Arsenic (As) is a toxic metalloid existing widely in the environment, and chronic exposure to it through contaminated drinking water has become a global problem of public health. The present study focused on the protective effects of selenium on oxidative damage of chronic arsenic poisoning in rat liver. Rats were divided into four groups at random and given designed treatments for 20 weeks. The oxidative damage of liver tissue was evaluated by lipid peroxidation and antioxidant enzymes. Oxidative stress related genes were detected to reflect the liver stress state at the molecular level. Compared to the control and Na₂SeO₃ groups, the MDA content in liver tissue was decreased and the activities of antioxidant enzymes were increased in the Na₂SeO₃ intervention group. The mRNA levels of SOD1, CAT, GPx and Txnrd1 were increased significantly (P < 0.05) in the combined Na₂SeO₃ + NaAsO₂ treatment group. The expressions of HSP70 and HO-1 were significantly (P < 0.05) increased in the NaAsO₂ group and reduced in the combined treatment group. The results indicate that long-term intake of NaAsO₂ causes oxidative damage in the rat liver, and Na₂SeO₃ protects liver cells by adjusting the expression of oxidative stress related genes to improve the activities of antioxidant enzymes.     

Influence of NADPH oxidase inhibition on oxidative stress parameters in rat hearts

BackgroundThe aim of this study was to assess whether apocynin, an nicotinamide adenine dinucleotide phosphate (NADPH) oxidase blocker, influences lipid peroxidation TBARS, hydrogen peroxide (H₂O₂) content, protein level, heart edema, tumor necrosis factor a (TNF-α) concentration or the glutathione redox system in heart homogenates obtained from endothelin 1 (ET-1)-induced oxidative stress rats.MethodsExperiments were carried out on adult male Wistar-Kyoto rats. The animals were divided into 4 groups: Group I: saline-treated control; Group II: saline followed by ET-1 (3 μg/kg b.w., iv); Group III: apocynin (5 mg/kg b.w., iv) administered half an hour before saline; Group IV: apocynin (5 mg/kg b.w., iv) administered half an hour before ET-1 (3 μg/kg b.w., iv).ResultsInjection of ET-1 alone showed a significant (p < 0.001) increase in thiobarbituric acid reactive substances (TBARS) and the hydrogen peroxide level (p < 0.01) vs. control, as well as a decrease (p < 0.001) in the GSH level. Apocynin significantly decreased TBARS (p < 0.001) and H₂O₂ (p < 0.05) level (vs. control) as well as improved protein level (p < 0.001) in the heart. Apocynin also prevented ET-1-induced heart edema (p < 0.05). The presence of ET-1 increased the concentration of TNF-α (p < 0.05) while apocynin decreased it (p < 0.05). Our results indicate that ET-1 may induce oxidative stress in heart tissue by reducing the GSH/GSSG ratio, stimulating lipid peroxidation and increasing TNF-α concentration. Apocynin diminished these measures of oxidative stress and TNF-α.ConclusionET-1-induced formation of ROS in the heart is at least partially regulated via NADPH oxidase.     

Dipeptidyl peptidase-4 inhibitor,linagliptin, ameliorates endothelial dysfunction and atherogenesis in normoglycemic apolipoprotein-E deficient mice

BackgroundDipeptidyl peptidase-4 (DPP-4) inhibitors have vasoprotective effects. This study investigated whether a recently approved DPP-4 inhibitor, linagliptin (Lina), suppresses atherogenesis in non-diabetic apolipoprotein-E deficient (ApoE^−/−) mice, and examined its effects on endothelial function.Methods and resultsLina (10 mg/kg/day) was administered orally to ApoE^−/− mice for 20 weeks. Lina reduced atherogenesis without alteration of metabolic parameters including blood glucose level compared with control (P < 0.05). Results of immunohistochemical analyses and quantitative RT-PCR demonstrated that Lina significantly decreased inflammatory molecule expression and macrophage infiltration in the atherosclerotic aorta. Lina administration to ApoE^−/− mice for 9 weeks ameliorated endothelium-dependent vasodilation compared with that in untreated mice. Plasma active glucagon-like peptide-1 (GLP-1) level was significantly higher in the treated group (P < 0.05). Exendin-4 (Ex-4), a GLP-1 analog, ameliorated endothelium-dependent vasodilation impaired by palmitic acid (PA) in wild-type mouse aortic segments. Ex-4 promoted phosphorylation of eNOS^Ser1177 and Akt, both of which were abrogated by PA, in human umbilical vein endothelial cells. In addition, Lina administration to ApoE^−/− mice decreased oxidative stress, as determined by urinary 8-OHdG secretion and NADPH oxidase subunit expression in the abdominal aorta.ConclusionLina inhibited atherogenesis in non-diabetic ApoE^−/− mice. Amelioration of endothelial dysfunction associated with a reduction of oxidative stress by GLP-1 contributes to the atheroprotective effects of Lina.     

A clinical comparison of the efficacy and safety of biosimilar G-CSF and originator G-CSF in haematopoietic stem cell mobilization

BackgroundRecombinant granulocyte colony-stimulating factor (G-CSF) is widely used to mobilize haematopoietic stem cells. We compared the efficacy and safety of a biosimilar G-CSF (Zarzio^®, Sandoz Biopharmaceuticals) with the originator G-CSF (Neupogen^®, Amgen) in patients with haematological malignancies.MethodsA total of 108 patients were included in this study, 59 of whom were female (49 male), with an overall median age of 51 years (range 19–69). Patients had multiple myeloma (n = 46), non-Hodgkin's lymphoma (n = 28), Hodgkin's lymphoma (n = 26), or other diagnosis (n = 8). After administration of mobilizing regimens (primarily high-dose etoposide, high-dose cyclophosphamide, intermediate-dose Ara-C or ESHAP), patients were randomized to a standard daily 10 μg/kg dose of biosimilar G-CSF (n = 54) or originator G-CSF (n = 54).ResultsMedian duration of G-CSF administration was 8 days with both biosimilar G-CSF (range 4–17) and originator G-CSF (range 4–14). Both groups had a median of one apheresis with a median time until first apheresis of 11 days. There were no statistically significant differences between groups in the mean ± SD number of mobilized CD34+ cells/μL in peripheral blood or the number of CD34+ cells/kg body weight. Five patients (9%) in the originator G-CSF group and six patients in the biosimilar G-CSF group (11%) did not mobilize sufficient CD34+ cells. The adverse event profile was similar between groups.ConclusionsA biosimilar G-CSF (Zarzio^®) demonstrated similar efficacy and safety as the reference originator G-CSF (Neupogen^®) in hematopoietic stem cell mobilization in patients with haematological malignancies.     

Bifenthrin-induced oxidative stress in human erythrocytes in vitro and protective effect of selected flavonols

Bifenthrin is a synthetic pyrethroid with a broad spectrum of insecticidal and acaricidal activity used to control wide range of insect pests in a variety of applications. This investigation was designed to examine (1) bifenthrin as an inducer of oxidative stress in human erythrocytes in vitro through effects on catalase (CAT) and superoxide dismutase (SOD) activities, and (2) the role of the flavonoids quercetin (Q, 40 and 80 μM) and rutin (R, 80 μM) in alleviating the effects of bifenthrin.Erythrocytes were divided into portions. The first portion was incubated for 4 h at 37 °C with different concentrations (0, 42.2, 211, 1055 ppm) of bifenthrin. The other portions were preincubated with Q or R for 30 min, followed incubation with bifenthrin for 4 h. The influence of solvent (ethanol) was also checked on the parameters studied. Malondialdehyde (MDA) concentrations, CAT and SOD activities were measured in all treatment portions of erythrocytes.Our results demonstrated that bifenthrin-induced oxidative stress causes enhanced lipid peroxidation and decreased antioxidative enzyme activities in human peripheral blood. R pretreated erythrocytes were protected against the increase of MDA induced by bifenthrin. Q (80 μM) and R pretreated erythrocytes were protected against the inhibition of CAT activity induced by bifenthrin.The protective action against the inhibition of SOD activity of Q was greater than that of R at the same concentration. These results suggest that Q and R may play a role in reducing bifenthrin-induced oxidative stress in vitro.     

Investigation into the role of Cu/Zn-SOD delivery system on its antioxidant and antiinflammatory activity in rat model of peritonitis

BackgroundThe current study evaluated the role of delivery system (solution, conventional liposomes and PEG-ylated liposomes) on superoxide dismutase (SOD) antioxidant and antiinflammatory properties in a rat model of lipopolysaccharide (LPS)-induced peritonitis.MethodsFifty male albino rats (Wistar-Bratislava) were divided into five groups (n = 10). Control group received saline and the other four groups received intraperitoneal injections of LPS (5 mg/kg). Among the LPS-injected groups, one was LPS control group and the other three groups received the endotoxin injection 30 min after receiving the same dose of SOD (500 U/kg, ip) in different delivery systems: saline solution (SOD-S), conventional liposomes (SOD-L) or PEG-ylated liposomes (SOD-PL). The animals were euthanized 6 h after LPS injection, blood samples were collected and acute phase response (total and differential leukocytes count; tumor necrosis factor α), antioxidants (total antioxidants; reduced glutathione), oxidative stress (total oxidants; lipid peroxidation) and nitrosative stress (nitric oxide metabolites; nitrotyrosine) were evaluated.ResultsIntraperitoneal administration of LPS to rats induced a marked inflammatory and oxidative response in plasma. On the other hand, all SOD formulations had protective effect against endotoxin-induced inflammation and oxidative/nitrosative stress, but PEG-ylated liposomes had the most significant activity. Thus, SOD-PL administration significantly reduced the effects of LPS on bone marrow acute phase response, the oxidative status and production of nitric oxide metabolites, while increasing the markers of antioxidant response in a significant manner.ConclusionSOD supplementation interferes both with inflammatory and oxidative pathways involved in LPS-induced acute inflammation, PEG-ylated liposomal formulation being of choice among the tested delivery systems.     

Potential role of licofelone,minocycline and their combination against chronic fatigue stress induced behavioral,biochemical and mitochondrial alterations in mice

BackgroundChronic fatigue stress (CFS) is a common complaint among general population. Persistent and debilitating fatigue severely impairs daily functioning and is usually accompanied by combination of several physical and psychiatric problems. It is now well established fact that oxidative stress and neuroinflammation are involved in the pathophysiology of chronic fatigue and related disorders. Targeting both COX (cyclooxygenase) and 5-LOX (lipoxygenase) pathways have been proposed to be involved in neuro-protective effect.MethodsIn the present study, mice were put on the running wheel apparatus for 6 min test session daily for 21 days, what produced fatigue like condition. The locomotor activity and anxiety like behavior were measured on 0, 8^th, 15^th and 22^nd day. The brains were isolated on 22^nd day immediately after the behavioral assessments for the estimation of oxidative stress parameters and mitochondrial enzyme complexes activity.ResultsPre-treatment with licofelone (2.5, 5 and 10 mg/kg, po) and minocycline (50 and 100 mg/kg, po) for 21 days, significantly attenuated fatigue like behavior as compared to the control (rotating wheel activity test session, RWATS) group. Further, licofelone (5 and 10 mg/kg, po) and minocycline (50 and 100 mg/kg, po) drug treatments for 21 days significantly attenuated behavioral alterations, oxidative damage and restored mitochondrial enzyme complex activities (I, II, III and IV) as compared to control, whereas combination of licofelone (5 mg/kg) with minocycline (50 mg/kg) significantly potentiated their protective effect which was significant as compared to their effect per se.ConclusionThe present study highlights the therapeutic potential of licofelone, minocycline and their combination against CFS in mice.     

Maturin acetate from Psacalium peltatum (Kunth) Cass. (Asteraceae) induces immunostimulatory effects in vitro and in vivo

Maturin acetate (MA) is one of main constituents in Psacalium peltatum. The cytotoxic effects of MA on tumorigenic cells were evaluated using the MTT assay. The in vitro immunostimulatory effects of maturin acetate (MA) were evaluated on the viability of murine splenocytes and macrophages, and human peripheral blood mononuclear cells (PBMC). The effects of MA on the production of nitrous oxide, pinocytosis and lysosomal enzyme activity were assayed in murine macrophages RAW 264.7. The effects of MA on the NK cell activity were also assayed. The in vivo immunostimulatory activities of MA were evaluated on BALB/c mice immunosuppressed with cyclophosphamide (CY). MA lacks cytotoxic activity against human cancer cells (IC₅₀ > 200 μM). In the absence of LPS, MA 10 μM or higher stimulated significantly (P ? 0.05), compared to untreated cells (-LPS), the viability of murine macrophages and splenocytes. In the absence of LPS, MA 10 μM or higher stimulated significantly (P ? 0.05), compared to untreated cells (-LPS), the lysosomal enzyme activity and pinocytosis. In immunosuppressed mice, MA increases significantly (P ? 0.05), compared to CY-treated mice, the production of IL-2 and IL-15 and IFN-γ. In conclusion, MA exerts immunostimulatory activities in vitro and in vivo.     

Maternal exposure to environmental tobacco smoke,GSTM1/T1 polymorphisms and oxidative stress

Environmental tobacco smoking (ETS) is known to be associated with adverse pregnancy outcomes. The purpose of this study was to investigate the relationship between maternal exposure to ETS and oxidative stress for neonates, as well as the effect of maternal genetic polymorphisms, glutathione-S-transferase M1 (GSTM1) and GSTT1, on this relationship.We used the radioimmunoassay to measure the urinary concentration of cotinine in 266 pregnant women who denied smoking cigarettes during pregnancy and in their singleton babies. In addition, the urinary concentration of malondialdehyde (MDA) and 8-hydroxy-2-deoxyguanosine (8-OH-dG) were assessed using high-performance liquid chromatography and enzyme-linked immunosorbent assay, respectively. We also extracted DNA from whole blood obtained from the mothers and then conducted polymerase chain reaction on the samples to determine the GSTM1 and GSTT1 genotypes.The maternal cotinine concentration was found to be significantly associated with the fetal cotinine concentration, particularly for mothers whose urine cotinine concentrations were above 120 μg/g cr (p < 0.01). The fetal urine cotinine concentration was also found to be significantly associated with the fetal urine MDA concentration (p < 0.01). When the null type maternal GSTM1 or the wild type GSTT1 was present, the maternal oxidative stress level increased significantly as the maternal continine concentration increased (MDA: p < 0.01; 8-OH-dG: p < 0.01). No significant relationships were found between maternal cotinine and fetal oxidative stress markers, however, the fetal MDA levels increased significantly as fetal cotinine levels increased.These results suggest that the maternal exposure to ETS affects the fetal urine cotinine concentration and induces production of maternal oxidative stress. In addition, maternal genetic polymorphisms of GSTM1 and GSTT1 may modify the oxidative stress by maternal exposure to ETS.     

TLR4-HMGB1 signaling pathway affects the inflammatory reaction of autoimmune myositis by regulating MHC-I

ObjectivesTo analyze the effects of TLR4 on the expression of the HMGB1, MHC-I and downstream cytokines IL-6 and TNF-α, and to investigate the biological role of the TLR4-HMGB1 signaling pathway in the development of the autoimmune myositis.MethodsWe built mice models with experimental autoimmune myositis (EAM) and used the inverted screen experiment to measure their muscle endurance; we also examined inflammatory infiltration of muscle tissues after HE staining; and we assessed the expression of MHC-I using immunohistochemistry. In addition, peripheral blood mononuclear cells (PBMC) were extracted and flow cytometry was utilized to detect the effect of IFN-γ on the expression of MHC-I. Furthermore, PBMCs were treated with IFN-γ, anti-TLR4, anti-HMGB1 and anti-MHC-I. Real-time PCR and western blotting were employed to examine the expressions of TLR4, HMGB1 and MHC-I in different groups. The ELISA method was also utilized to detect the expression of the downstream cytokines TNF-α and IL-6.ResultsThe expressions of TLR4, HMGB1 and MHC-I in muscle tissues from mice with EAM were significantly higher than those in the control group (all P < 0.05). After IFN-γ treatment, the expressions of TLR4, HMGB1, MHC-I, TNF-α and IL-6 in PBMCs significantly increased (all P < 0.05). The treatment of anti-TLR4, anti-HMGB1 and anti-MHC-I could significantly downregulate the expression of MHC-I (all P < 0.05). In addition, anti-TLR4 and anti-HMGB1 significantly reduced the expression of TNF-α and IL-6 (all P < 0.05).ConclusionsThe TLR4-HMGB1 signaling pathway affects the process of autoimmune myositis inflammation by regulating the expression of MHC-I and other pro-inflammatory cytokines.