iNOS-positive Cells Research Articles

Abstract 133: Triggering Receptor Expressed on Myeloid Cells-2 Improves Post-stroke Neurological Function and Regulates Microglia/Macrophage Phenotype in Aged Female Mice

Background and Purpose: We previously reported that triggering receptor expressed on myeloid cells-2 (TREM2) expression on microglia/macrophage activates its phagocytic function and promotes stroke recovery. Although its importance has been demonstrated, it is unclear whether there is a similar effect in females. Furthermore, TREM2 mutations have been linked to dementia. In the present study, we explored the influence of TREM2 deficiency in aged female mice subjected to middle cerebral artery occlusion (MCAO). We assessed histology, sensorimotor and cognitive functions along with post stroke immune responses. Methods: Age matched (≥8months) female TREM2 knockout (TREM2-KO) and Wild type (Wt) mice were subjected to permanent distal MCAO. Sensorimotor and cognitive (Y-maze test) functions and infarct size were assessed up to 14d after stroke (n=6/each). Microglial activation (isolectin B4, IB4 & CD68), phagocytosis (oil red O stain) and M1 (NO/iNOS) and M2 (arginase, Arg) markers 1,3,7 and 14d post MCAO. Results: At baseline, aged female mice lacking TREM2 had similar cognitive & sensorimotor function. However, post stroke, TREM2-KO females, compared to Wt, showed worsened cognitive (p<0.01) and neurological function (p<0.05). Female TREM2 knockout mice also had larger infarct volumes (p<0.05) and reduced phagocytic activity (p<0.01). Microglia/macrophage accumulation, which peaked by 7d, was significantly inhibited in TREM2-KO mice (p<0.01). TREM2-KO mice also showed fewer CD68+ phagocytes; however, iNOS positive CD68 cells were significantly increased at days 3 & 7 (p<0.05), while, Arg positive CD68 cells was significantly decreased at days 7 & 14 (p<0.05) in the TREM2-KO group, compared to the Wt group and suggests a M2 bias for TREM2. Conclusions: Similar to male mice, TREM2 seems to have beneficial properties in female mice. TREM2 deficient female mice had larger infarct sizes and worsened neurological performance. Microglial responses also seemed to suggest that TREM2 promotes beneficial and/or recovery pathways.

Abstract WP134: Calcium Release-activated Calcium Channel Inhibition Improves Neurological Outcome and Suppresses Inflammation in Female Mice Following Cerebral Ischemic Injury

Purpose: Calcium release-activated calcium (CRAC) channels activate microglia via store-operated Ca 2+ entry. We previously reported that the specific CRAC channel inhibitor (CM-EX-137) had neuroprotective effects in a model of experimental stroke due to suppression of microglial activation in male mice. However, it is well known that sex differences can influence stroke outcome as well as immune responses. Whether CRAC channel inhibition in female mice is similarly neuroprotective is unknown. The purpose of this study is to determine whether female mice respond similarly CRAC channel inhibition in a stroke model Subjects and Methods: C57/BL6 female mice, aged 2 months were subjected to distal middle cerebral artery occlusion (dMCAO) were treated with CM-EX-137 (5mg/kg IP; dMCAO-CM) or vehicle control (dMCAO-vehicle) daily for 3d. Stroke surgeries were carried out during the same phase of the estrus cycle (diestrus) to avoid potential confounds due to varying estrogen levels. These groups were also compared to sham groups. CM-EX-137 was injected immediately after occlusion and at 24 & 48h. Brains were extracted 3d post dMCAO. Neurological functions were evaluated before surgery, and 1 & 3d later. Lesion size was estimated from cresyl violet stains. Microglia and astrocyte activation were assessed using immunohistochemistry. Results: Neurological function at both 24 and 72 h post ischemia were significantly improved in dMCAO-CM group, compared with dMCAO-vehicle group (n=7/group; p<0.001). CM-EX-137 also significantly reduced infarct volume at 3dafter dMCAO (p<0.001). CM-EX-137 inhibited microglial/monocyte/macrophage activation (p<0.001; isolectin B4, CD68) and led to decreased iNOS positive cells (p<0.01). CM-EX-137 also decreased expression of CRAC channel components STIM1 and Orai1 on CD68+ macrophages (p<0.001). However, GFAP staining indicated that CM-EX-137 had no effect on astrocyte activation. Conclusion: CRAC channel inhibition reduce inflammatory responses and improve neurological outcome following experimental stroke in female animals. These results are in line with those that we observed in male mice. Thus, CRAC channel inhibition seems to be similarly effective and anti-inflammatory in both genders in a stroke model.

Suppressed microRNA-96 inhibits iNOS expression and dopaminergic neuron apoptosis through inactivating the MAPK signaling pathway by targeting CACNG5 in mice with Parkinson\u2019s disease

BackgroundThere have been a number of reports implicating the association of microRNAs (miRs) and the MAPK signaling pathway with the dopaminergic neuron, which is involved in the development of Parkinson’s disease (PD). The present study was conducted with aims of exploring the role of miR-96 in the activation of iNOS and apoptosis of dopaminergic neuron through the MAPK signaling pathway in mice with PD.MethodsThe miR and the differentially expressed gene in PD were screened out and the relationship between them was verified. A mouse model of PD induced by MPTP and was then constructed and treated with miR-96 mimic/inhibitor and CACNG5 overexpression plasmid to extract nigral dopaminergic neuron for the purpose of detecting the effect of miR-96 on PD. The TH and iNOS positive neuronal cells, the apoptotic neuronal cells by TUNEL staining, and expression of miR-96, CACNG5, iNOS, p38MAPK, p-p38MAPK, c-Fos, Bax, and Bcl-2 in substantia nigra dopaminergic neuronal tissues were evaluated.ResultsThe results obtained from the aforementioned procedure were then verified by cell culture of the SH-SY5Y cells, followed by treatment with miR-96 mimic/inhibitor, CACNG5 overexpression plasmid and the inhibitor of the MAPK signaling pathway. CACNG5 was confirmed as a target gene of miR-96. The inhibition of miR-96 resulted in a substantial increase in nigral cells, TH positive cells and expression of CACNG5 and Bcl-2 in nigral dopaminergic neuronal tissues, and a decrease in iNOS positive cells, apoptotic neuronal cells, and expression of iNOS, p38MAPK, p-p38MAPK, c-Fos, and Bax.ConclusionThe above results implicated that the downregulation of miR-96 inhibits the activation of iNOS and apoptosis of dopaminergic neuron through the blockade of the MAPK signaling pathway by promoting CACNG5 in mice with PD.

Protective Effects of Anti-IL17 on Acute Lung Injury Induced by LPS in Mice.

Introduction: T helper 17 (Th17) has been implicated in a variety of inflammatory lung and immune system diseases. However, little is known about the expression and biological role of IL-17 in acute lung injury (ALI). We investigated the mechanisms involved in the effect of anti-IL17 in a model of lipopolysaccharide (LPS)-induced acute lung injury (ALI) in mice.Methods: Mice were pre-treated with anti-IL17, 1h before saline/LPS intratracheal administration alongside non-treated controls and levels of exhaled nitric oxide (eNO), cytokine expression, extracellular matrix remodeling and oxidative stress, as well as immune cell counts in bronchoalveolar lavage fluid (BALF), and respiratory mechanics were assessed in lung tissue.Results: LPS instillation led to an increase in multiple cytokines, proteases, nuclear factor-κB, and Forkhead box P3 (FOXP3), eNO and regulators of the actomyosin cytoskeleton, the number of CD4+ and iNOS-positive cells as well as the number of neutrophils and macrophages in BALF, resistance and elastance of the respiratory system, ARG-1 gene expression, collagen fibers, and actin and 8-iso-PGF2α volume fractions. Pre-treatment with anti-IL17 led to a significant reduction in the level of all assessed factors.Conclusions: Anti-IL17 can protect the lungs from the inflammatory effects of LPS-induced ALI, primarily mediated by the reduced expression of cytokines and oxidative stress. This suggests that further studies using anti-IL17 in a treatment regime would be highly worthwhile.

Immunohistochemical localization of nitric oxide synthase (NOS) isoforms in epidermis and gill epithelium of an angler catfish, Chaca chaca (Siluriformes, Chacidae)

The present study is concerned with the expression and localization of nitric oxide synthase (NOS) isoforms, nNOS, eNOS and iNOS in the epidermis and the gill epithelium of Chaca chaca by means of immunohistochemical techniques. nNOS immunoreactivity was observed in the outer layer epithelial cells of the epidermis, outer epithelium of gill filaments at their distal regions and in between the secondary lamellae. iNOS positive cells were observed at intervals in the epidermis from basal layer to superficial layer, in outer layers of epithelium of the gill filament and in epithelium of the secondary lamellae. The expression of eNOS is similar to that of iNOS in the gills. In addition, NOS activity was also observed in the taste buds in the epidermis. The expression of different NOS isoforms in C. chaca are associated to increase the adaptability and survivability of the fish in hypoxic condition, help in defence and ion regulation and sensory functions. The study could be useful to understand the expression of NOS isoforms in different fish tissues and their diverse role in relation to the physiology of the fish.

Low dose of chlorine exposure exacerbates nasal and pulmonary allergic inflammation in mice

Work-exacerbated asthma (WEA) is defined as preexisting asthma that worsens with exposure to irritants [e.g., chlorine (Cl2) derivatives] in the workplace. The maximum allowable concentration in the workplace of Cl2 exposure is 3 mg/ m3 (described in OSHA). We investigated in an experimental asthma model in mice the effects of a single exposure to a sodium hypochlorite dose with this allowed chlorine concentration and a tenfold higher dose. Acute chlorine exposure at 3.3 mg/m3 in the OVA-sensitized group increased eosinophils in the peribronquial infiltrate, cytokine production, nasal mucus production and the number of iNOS positive cells in the distal lung compared to only sensitized mice. The exposure to a higher dose of 33.3 mg/m3 in the OVA-sensitized group resulted in an increase in respiratory system elastance, in the total and differential numbers of inflammatory cells in bronchoalveolar lavage fluid, IL-4, IL-5, and IL-17 in the lungs, eosinophils in peribronquial infiltrate and mucus content in nasal compared to non-exposed and sensitized animals. In this asthma model, chorine exposures at an allowable dose, contributed to the potentiation of Th2 responses. The functional alterations were associated with increased iNOS and ROCK-2 activation in the distal lung.

A Natural Dietary Supplement with a Combination of Nutrients Prevents Neurodegeneration Induced by a High Fat Diet in Mice.

Obesity and metabolic disorders can be risk factors for the onset and development of neurodegenerative diseases. The aim of the present study was to investigate the protective effects of a natural dietary supplement (NDS), containing Curcuma longa, silymarin, guggul, chlorogenic acid and inulin, on dysmetabolism and neurodegeneration in the brains of high fat diet (HFD)-fed mice. Decrease in the expression of FACL-4, CerS-1, CerS-4, cholesterol concentration and increase in the insulin receptor expression and insulin signaling activation, were found in brains of NDS-treated HFD brains in comparison with HFD untreated-mice, suggesting that NDS is able to prevent brain lipid accumulation and central insulin resistance. In the brains of NDS-treated HFD mice, the levels of RNS, ROS and lipid peroxidation, the expression of p-ERK, H-Oxy, i-NOS, HSP60, NF-kB, GFAP, IL-1β, IL-6 and CD4 positive cell infiltration were lower than in untreated HFD mice, suggesting antioxidant and anti-inflammatory effects of NDS. The decreased expression of p-ERK and GFAP in NDS-treated HFD mice was confirmed by immunofluorescence. Lastly, a lower number of apoptotic nuclei was found in cortical sections of NDS-treated HFD mice. The present data indicate that NDS exerts neuroprotective effects in HFD mice by reducing brain fat accumulation, oxidative stress and inflammation and improving brain insulin resistance.

Inhibition of Nitric Oxide Production in Activated Macrophages Caused by Toxoplasma gondii Infection Occurs by Distinct Mechanisms in Different Mouse Macrophage Cell Lines.

Toxoplasma gondii, the causative agent of toxoplasmosis, is a widespread intracellular parasite able to infect virtually any nucleated cell. T. gondii infection of activated macrophages inhibits nitric oxide (NO) production; however, parasite effectors responsible for this block have not been defined. Macrophage populations are extremely heterogeneous, responding differently to stimuli and to parasite infection. Here we evaluated the inhibition of NO production caused by T. gondii infection of J774-A1 and RAW 264.7 macrophages and assessed the role of several known parasite virulence factors in this phenotype. Infection of activated macrophages from both macrophage lines reduced NO production, however, the mechanism of this decrease was different. Consistent with previous reports, infected J774-A1 macrophages had reduced iNOS expression and lower number of iNOS positive cells. In contrast, T. gondii infection of RAW 264.7 macrophages did not alter iNOS expression or the number of iNOS positive cells, and yet it led to lower levels of NO production. Deletion of a number of previously defined virulence factors including ROP kinases that disrupt innate immune factors, TgIST which blocks STAT1 activation, as well as the secretory trafficking proteins ASP5 and MYR1, did not alter the phenotype of decreased NO production. Taken together our findings indicate that T. gondii infection inhibits NO production of activated macrophages by different mechanisms that involve reduction of iNOS expression vs. iNOS impairment, and suggest that a novel parasite effector is involved in modulating this important host defense pathway.

Laquinimod treatment in the R6/2 mouse model

The transgenic mouse model R6/2 exhibits Huntington’s disease (HD)-like deficits and basic pathophysiological similarities. We also used the pheochromocytoma-12 (PC12)-cell-line-model to investigate the effect of laquinimod on metabolic activity. Laquinimod is an orally administered immunomodulatory substance currently under development for the treatment of multiple sclerosis (MS) and HD. As an essential effect, increased levels of BDNF were observed. Therefore, we investigated the therapeutic efficacy of laquinimod in the R6/2 model, focusing on its neuroprotective capacity. Weight course and survival were not influenced by laquinimod. Neither were any metabolic effects seen in an inducible PC12-cell-line model of HD. As a positive effect, motor functions of R6/2 mice at the age of 12 weeks significantly improved. Preservation of morphologically intact neurons was found after treatment in the striatum, as revealed by NeuN, DARPP-32, and ubiquitin. Biochemical analysis showed a significant increase in the brain-derived neurotrophic factor (BDNF) level in striatal but not in cortical neurons. The number of mutant huntingtin (mhtt) and inducible nitric oxide synthase (iNOS) positive cells was reduced in both the striatum and motor cortex following treatment. These findings suggest that laquinimod could provide a mild effect on motor function and striatal histopathology, but not on survival. Besides influences on the immune system, influence on BDNF-dependent pathways in HD are discussed.

Plant Proteinase Inhibitor BbCI Modulates Lung Inflammatory Responses and Mechanic and Remodeling Alterations Induced by Elastase in Mice.

Background. Proteinases play a key role in emphysema. Bauhinia bauhinioides cruzipain inhibitor (BbCI) is a serine-cysteine proteinase inhibitor. We evaluated BbCI treatment in elastase-induced pulmonary alterations. Methods. C57BL/6 mice received intratracheal elastase (ELA group) or saline (SAL group). One group of mice was treated with BbCI (days 1, 15, and 21 after elastase instillation, ELABC group). Controls received saline and BbCI (SALBC group). After 28 days, we evaluated respiratory mechanics, exhaled nitric oxide, and bronchoalveolar lavage fluid. In lung tissue we measured airspace enlargement, quantified neutrophils, TNFα-, MMP-9-, MMP-12-, TIMP-1-, iNOS-, and eNOS-positive cells, 8-iso-PGF2α, collagen, and elastic fibers in alveolar septa and airways. MUC-5-positive cells were quantified only in airways. Results. BbCI reduced elastase-induced changes in pulmonary mechanics, airspace enlargement and elastase-induced increases in total cells, and neutrophils in BALF. BbCI reduced macrophages and neutrophils positive cells in alveolar septa and neutrophils and TNFα-positive cells in airways. BbCI attenuated elastic and collagen fibers, MMP-9- and MMP-12-positive cells, and isoprostane and iNOS-positive cells in alveolar septa and airways. BbCI reduced MUC5ac-positive cells in airways. Conclusions. BbCI improved lung mechanics and reduced lung inflammation and airspace enlargement and increased oxidative stress levels induced by elastase. BbCI may have therapeutic potential in chronic obstructive pulmonary disease.

Immunohistochemical evaluation of inducible nitric oxide synthase in the epithelial lining of odontogenic cysts: A qualitative and quantitative analysis.

Introduction:The three common odontogenic cysts include radicular cysts (RCs), dentigerous cysts (DCs), and odontogenic keratocysts (OKCs). Among these 3 cysts, OKC is recently been classified as benign keratocystic odontogenic tumor attributing to its aggressive behavior, recurrence rate, and malignant potential. The present study involved qualitative and quantitative analysis of inducible nitric oxide synthase (iNOS) expression in epithelial lining of RCs, DCs, and OKCs, compare iNOS expression in epithelial linings of all the 3 cysts and determined overexpression of iNOS in OKCs which might contribute to its aggressive behavior and malignant potential.Aims:The present study is to investigate the role of iNOS in the pathogenesis of OKCs, DCs, and RCs by evaluating the iNOS expression in the epithelial lining of these cysts.Subjects and Methods:Analysis of iNOS expression in epithelial lining cells of 20 RCs, 20 DCs, and 20 OKCs using immunohistochemistry done.Statistical Analysis Used:The percentage of positive cells and intensity of stain was assessed and compared among all the 3 cysts using contingency coefficient. Kappa statistics for the two observers were computed for finding interobserver agreement.Results:The percentage of iNOS-positive cells was found to be remarkably high in OKCs (12/20) –57.1% as compared to RCs (6/20) – 28.6% and DCs (3/20) – 14.3%. The interobserver agreement for iNOS-positive percentage cells was arrived with kappa values with OKCs → Statistically significant (P > 0.000), RCs → statistically significant (P > 0.001) with no significant values for DCs. No statistical difference exists among 3 study samples in regard to the intensity of staining with iNOS.Conclusions:Increased iNOS expression in OKCs may contribute to bone resorption and accumulation of wild-type p53, hence, making OKCs more aggressive.

The effects of prenatal "psychological" stressor exposure on lung inflammation and hyperresponsiveness in adult rat offspring.

The aim of this study was to establish whether exposure of pregnant rats to uncontrollable (psychological) stressors might change the likelihood of their offspring to exhibit functional and histopathological abnormalities suggestive of asthma in adulthood. Pregnant rats (n = 16) underwent one of three treatments: electric shocks of a maximum duration of 10 s that could be escaped (controllable group; C) those that could not be escaped (uncontrollable group; U) or no shocks (control group; N). The offspring (n = 54) were kept in animal house under standard conditions until 3 months of age, when lung hyperresponsiveness, histopathology, immunohistochemical measurements of the cytokines interleukin (IL) 2, IL-4, IL-5, and IL-13 and actin as well as oxidative stress based on iNOS-positive cell counts and isoprostane PGF2α contents were assessed. The results showed that prenatal exposure to physical stressors (shocks) caused lung hyperresponsiveness and increased cytokine expression; exposure to uncontrollable shock (group U) had a differential effect on the expression of IL-2, IL-5, and IL-13 in inflammatory cells compared to exposure to controllable shock (group C), which characterizes the "psychological" aspect of stress. The results show that not only stress but also its uncontrollability during gestation might increase the likelihood that the offspring will exhibit functional and histopathological abnormalities suggestive of asthma. These findings strengthen the importance of psychological control with regard to environmental stimuli for the occurrence of several illnesses, suggesting the desirability of integration among various fields of science.

Effects of alpha-tocopherol on gingival expression of inducible nitric oxide synthase in the rats with experimental periodontitis and diabetes.

The aim of this study was to investigate effects of α-tocopherol and/or insulin on the number of gingival inducible nitric oxide synthase (iNOS) positive cells in rats with experimental periodontitis with or without streptozotocin (STZ)-induced diabetes. A total of 60 Sprague-Dawley rats were divided into three groups: Group I: The group without diabetes; Group II: The group with STZ-induced diabetes; Group III: The group with STZ-induced diabetes receiving insulin therapy. All animals received anesthesia, and 3/0 silk suture was inserted around the mandibular molar teeth. All groups were divided into subgroups receiving saline (Groups IA, IIA, IIIA) and α-tocopherol injection (Groups IB, IIB, IIIB). After a period of 3 weeks, all rats were sacrificed, and the number of gingival iNOS positive cells was analyzed using image analysis software. Applying α-tocopherol suppressed the number of gingival iNOS positive cells in Groups IB, IIB, and IIIB compared to application of saline (Groups IA, IIA, and IIIA) (P < 0.05). Numbers of gingival iNOS positive cells were found to be similar in the Groups I and III (P > 0.05). Within limitations of the current study, this is the first study one may suggest that α-tocopherol may reduce oxidative damage in the gingiva of the rats with periodontitis with or without STZ-induced diabetes and increase effects of insulin.

Effects of catecholaminergic nerve lesion on endometrial development during early pregnancy in Mice.

Maternal stress is common during pregnancy and the postnatal period. This stress typically activates the sympathetic nervous system which releases catecholamines. This study explored the influence of sympathectomy by using neurotoxin 6-hydroxydopamine (6-OHDA) on embryo implantation, and investigated the influence mechanism of sympathectomy on reconstruction of endometrial structure during early pregnancy. In the 6-OHDA-treated mice, uterine glands in the endometrium developed poorly, and the gland epithelia were arranged irregularly during early pregnancy. Furthermore, vacuoles, karyopykosis and plasmarrhexis appeared in some gland epithelia. The percentage of uterine glands and the density of proliferating cell nuclear antigen (PCNA) positivity were dramatically decreased, and Fas ligand (FasL) expression was decreased in cells from pregnancy days 5-9 (E5-9) in the treated group. Antioxidant enzyme activity levels in uteri were lower but the malondialdehyde (MDA) levels were higher in the 6-OHDA mice than those in the control mice at E5-9. Similarly, the number of inducible nitric oxide synthase (iNOS) positive cells was significantly increased during early pregnancy following treatment with 6-OHDA. Our results have indicated that peripheral catecholaminergic nerve lesions induced by 6-OHDA cause adverse pregnancy outcomes through disruption of endometrial gland development, which increases oxidative stress and iNOS expression in the endometrium. Thus, catecholaminergic nerves might favourably influence blastocyst implantation, foetal survival and development during early pregnancy by oxidative state regulation and endometrial gland reconstruction.

Rat bone marrow-derived dendritic cells generated with GM-CSF/IL-4 or FLT3L exhibit distinct phenotypical and functional characteristics.

Dendritic cells are professional APCs that play a central role in the initiation of immune responses. The limited ex vivo availability of dendritic cells inspires the widespread use of bone marrow-derived dendritic cells as an alternative in research. However, the functional characteristics of bone marrow-derived dendritic cells are incompletely understood. Therefore, we compared functional and phenotypic characteristics of rat bone marrow-derived dendritic cells generated with GM-CSF/IL-4 or FLT3 ligand bone marrow-derived dendritic cells. A comparison of surface markers revealed that FLT3 ligand-bone marrow-derived dendritic cells expressed signal regulatory protein α, CD103, and CD4 and baseline levels of MHC class II, CD40, and CD86, which were highly up-regulated upon stimulation. Conversely, GM-CSF/IL-4-bone marrow-derived dendritic cells constitutively expressed signal regulatory protein α, CD11c, and CD11b but only mildly up-regulated MHC class II, CD40, or CD86 following stimulation. Expression of dendritic cell-associated core transcripts was restricted to FLT3 ligand-bone marrow-derived dendritic cells . GM-CSF/IL-4-bone marrow-derived dendritic cells were superior at phagocytosis but were outperformed by FLT3 ligand-bone marrow-derived dendritic cells at antigen presentation and T cell stimulation in vitro. Stimulated GM-CSF/IL-4-bone marrow-derived dendritic cells secreted more TNF, CCL5, CCL20, and NO, whereas FLT3 ligand-bone marrow-derived dendritic cells secreted more IL-6 and IL-12. Finally, whereas GM-CSF/IL-4-bone marrow-derived dendritic cell culture supernatants added to resting T cell cultures promoted forkhead box p3(+) regulatory T cell populations, FLT3 ligand-bone marrow-derived dendritic cell culture supernatants drove Th17 differentiation. We conclude that rat GM-CSF/IL-4-bone marrow-derived dendritic cells and FLT3 ligand-bone marrow-derived dendritic cells are functionally distinct. Our data support the current rationale that FLT3 ligand-bone marrow-derived dendritic cells mostly resemble classic dendritic cells but comprise additional minor subpopulations, whereas GM-CSF/IL-4-bone marrow-derived dendritic cells resemble monocyte-derived inflammatory dendritic cells (iNOS-positive monocyte-derived cells).

Different dose-dependent effects of ebselen in sciatic nerve ischemia-reperfusion injury in rats.

Ebselen is an organoselenium compound which has strong antioxidant and anti-inflammatory effects. We investigated the neuroprotective role of ebselen pretreatment in rats with experimental sciatic nerve ischemia-reperfusion (I/R) injury. Adult male Sprague Dawley rats were divided into four groups (N = 7 in each group). Before sciatic nerve I/R was induced, ebselen was injected intraperitoneally at doses of 15 and 30 mg/kg. After a 2 h ischemia and a 3 h reperfusion period, sciatic nerve tissues were excised. Tissue levels of malondialdehyde (MDA) and nitric oxide (NO), and activities of superoxide dismutase (SOD), glutathione peroxidase (GPx), and catalase (CAT) were measured. Sciatic nerve tissues were also examined histopathologically. The 15 mg/kg dose of ebselen reduced sciatic nerve damage and apoptosis (p<0.01), levels of MDA, NO, and inducible nitric oxide synthase (iNOS) positive cells (p<0.01, p<0.05, respectively), and increased SOD, GPx, and CAT activities (p<0.001, p<0.01, p<0.05, respectively) compared with the I/R group that did not receive ebselen. Conversely, the 30 mg/kg dose of ebselen increased sciatic nerve damage, apoptosis, iNOS positive cells (p<0.01, p<0.05, p<0.001) and MDA and NO levels (p<0.05, p<0.01) and decreased SOD, GPx, and CAT activities (p<0.05) compared with the sham group. The results of this study suggest that ebselen may cause different effects depending on the dose employed. Ebselen may be protective against sciatic nerve I/R injury via antioxidant and antiapoptotic activities at a 15 mg/kg dose, conversely higher doses may cause detrimental effects.

A plant proteinase inhibitor from Crataeva tapia (CrataBL) attenuates elastase-induced pulmonary inflammatory, remodeling, and mechanical alterations in mice

Chronic obstructive pulmonary disease (COPD) can lead to chronic and obstructive bronchitis and emphysema resulting in decreased bronchial lumen size.This study evaluated the effect of CrataBL, a protein isolated from the bark of Crataeva tapia, on lung mechanics, inflammation, and remodeling after elastase-induced pulmonary alterations in mice.The use of CrataBL led to decreased mechanical alterations, alveolar septum disruption (Lm), number of macrophage and neutrophil cells in the BALF, and TNF-α, MMP-9, MMP-12, TIMP-1, eNOS, and iNOS positive cells in the airways and alveolar walls compared to the animals in the ELA group. Moreover, a reduction in MUC-5-positive cells in the airway walls was observed. In conclusion, CrataBL attenuates changes in lung mechanics, inflammation, extracellular lung remodeling, and oxidative stress responses induced by the administration of elastase and decreased the volume fraction of isoprostane, collagen, and elastic fibers in the airways and alveolar walls compared to the animals in the ELA groups. Therefore, CrataBL is a potential therapeutic tool in the treatment of COPD.

ADIOL protects against 3-NP-induced neurotoxicity in rats: Possible impact of its anti-oxidant, anti-inflammatory and anti-apoptotic actions.

Huntington's disease (HD) is a progressive neurodegenerative disorder with a wide spectrum of cognitive, behavioral and motor abnormalities. The mitochondrial toxin 3-nitropropionic acid (3-NP) effectively induces specific behavioral changes and selective striatal lesions similar to that observed in HD. Some neurosteroids, synthesized in neurons and glial cells, previously showed neuroprotective abilities. 5-Androstene-3β-17β-diol (ADIOL) is a major metabolite of dehydroepiandrosterone (DHEA) with previously reported anti-inflammatory, anti-apoptotic and neuroprotective activities. The neuroprotective potential of ADIOL in HD was not previously investigated. Therefore, the present study investigated the neuroprotective effects of ADIOL against 3-NP-induced behavioral changes, oxidative stress, inflammation and apoptosis. Intraperitoneal administration of 3-NP (20mg/kg) for 4 consecutive days in rats caused significant loss in body weight, reduced prepulse inhibition (PPI) of acoustic startle response, locomotor hypoactivity with altered cortical/striatal histological structure, increased cortical/striatal oxidative stress, inflammation and apoptosis. Administration of ADIOL (25mg/kg, s.c.) for two days before 3-NP significantly attenuated the reduction in body weights and PPI, increased locomotor activity and restored cortical/striatal histological structure nearly to normal. Moreover, it displayed anti-oxidant, anti-inflammatory and anti-apoptotic activities as evidenced by the elevation of cortical and striatal reduced glutathione levels, reductions of cortical and striatal malondialdehyde, striatal tumor necrosis factor alpha and interleukin-6 levels. Only a small number of iNOS and caspase-3 positive cells were detected in sections from rats pretreated with ADIOL. This study suggests a potential neuroprotective role of ADIOL against 3-NP-induced Huntington's disease-like manifestations. Such neuroprotection can be attributed to its anti-oxidant, anti-inflammatory and anti-apoptotic activities.

Protective Effect of Curcumin versus N-acetylcystein on Acetaminophen Induced Hepatotoxicity in Adult Albino Rats

Curcumin exerted hepatoprotective influences on various animal models of liver injury such as carbon tetrachloride, endotoxin and thioacetamide. This Study aimed to investigate the protective effects of curcumin as compared to N-acetylcysteine in the rat model of acetaminophen induced hepatotoxicity. 55 albino rats were divided into four groups: group )control group(. Group II received acetaminophen. Group III received both acetaminophen and N-acetylcysteine and Group IV received both acetaminophen and curcumin. At the end of the experiment, liver specimens were processed for histological study by light microscope and stained immunohistochemically for detection of apoptosis in hepatic cells by using anti p53 and the oxidative damage by anti-inducible nitric oxide synthase (anti iNOS). Serum levels of liver injury markers were assessed as well as the levels of malondialdehyde (MDA), glutathione (GSH) and superoxide dismutase (SOD) activity were determined in all dissected tissues. Histological examination of liver sections of acetaminophen treated group revealed degeneration, cytoplasmic vacuolation and hydropic necrosis of hepatocytes. The central and the portal veins were dilated and congested and invading infiltrative inflammatory cells were appeared in association with significant increase in p53 and iNOS positive cells. Significant rise in serum levels of liver injury markers and MDA in liver tissues were recorded. However, levels of GSH and SOD were significantly decreased. Both curcumin and N-acetylcysteine resolved most of these morphological, immunohistochemical and biochemical alterations. Curcumin has protective effect similar to N-acetylcysteine against liver damage induced by acetaminophen in rats by reducing oxidative stress and apoptosis.

Inulin Diet Intervention on Chemopreventive and Inflammatory Markers in Tumorigenesis of Colorectal Cancer

Abstract The aim of this experiment was to investigate the infl uence of inulin administration on chemopreventive and inflammatory markers in dimethylhydrazine induced colorectal cancer development in rats. A group of 30 Sprague-Dawley rats was divided into a control group (CG), a group with dimethylhydrazine (DMH), and a group given dimethylhydrazine combined with the prebiotic (DMH+PRE). Dimethylhydrazine injection significantly (p&lt;0.001) elevated the immunoreactivity chemopreventive markers COX-2, NFκB, iNOS, elevated serum and jejunal mucosa levels of proinfl ammatory cytokine IL-2, and decreased serum and jejunal mucosa levels of regulatory cytokine IL-10. Inulin diet intervention significantly suppressed immunoreactivity of COX- 2, NFκB, iNOS positive cells in the tunica mucosae and tela submucosae of rat colon tissue, increased levels of IL-2 and decreased levels of IL-10. By determining the chemopreventive markers COX-2, iNOS and NFkB, which can be characterized as inflammatory markers, we confirmed the presence of inflammation in the colon as the number of COX-2, NFkB and iNOS immunoreactive cells was significantly higher after DMH application than in the control group. These findings indicate that dietary intake of inulin suppressed the expression of the observed markers, which play an important role in carcinogenesis and inflammation, which predispose the use of inulin in the prevention or treatment of human chronic diseases and its use as a nutritional supplement in veterinary medicine.