Cytokines IL-1β Levels Research Articles

Activation of TLRs/NF - κ B signal pathway and occurrence of different functionalcytokines during invasive pulmonary aspergillosis

Objective To study the activation of TLRs/NF-κBsignal pathway and production of different functional cytokines during invasive pulmonaryaspergillosis( IPA) , in order to probe the pathogene-sis of IPA. Methods Mouse wererandomly divided into normal, normal + inoculated with Aspergillus fumigatus( normalinoculation group), and immune suppression + inoculation with Aspergillus fumigatus(IPAmodel group) , the mouse were killed at different time points after inhalingAspergillus fumigatus spores by nose. Removing the lung tissue in a sterile manner andmaking pathological section respectively, counting Aspergillus fumigatus colony,dynamiclly detecting the expression of TLR2, TLR4 mRNA, variation of NF-κB p65protein, pro-inflammatory cytokines TNF-α, IL-1β and anti-inflammatory cytokines IL-10 levels in the lung tissue byRT-PCR and Western blot method during Aspergillus fumigatus infection in mouse. Results(1) When it's 72 h after inhaling Aspergillus fumigatus by nose, IPA model emerged severelung tissue inflammation, and generated a large number of hyphae, meanwhile, burthen ofAspergillus fumigatus was higher than normal inoculation group at each time point.(2)Compared with the normal inoculation group, IPA group whose TLR2 mRNA was lowexpression at early stage of infection (24 h), and emerged high expression at late stageof infection (120 h, 144 h); and TLR4 mRNA has been at a state of low expression in theinfection process; NF-κB p65 suddenly increased at early stage ofinfection(24 h) and then continued to decline. (3) After infected by Aspergillus fumigatusin normal mouse, proinflammatory cytokine TNF-α, IL-1β in lung exhibited high expression at the early stages of infection,and the highest expression levels appeared at 48 h or 72 h, then decreased and recoveredto normal level. And the expression level of anti-inflammatory cytokine IL-10 rised atlate stage of infection; The IP A mouse released a lot of anti-inflammatory cytokine IL-10at early stage of infection, which significantly reduced at late stage, and releasedpro-inflammatory cyto-kines TNF-α, IL-1β at slow and low level. Conclusion The abnormal activation ofTLRs/NF-β signaling pathway caused the loss of dynamic balance between pro-inflammatorycytokines and anti-inflammatory cytokines, leading to the occurrence and development ofIPA.

Biphasic Effect of Phyllanthus emblica L. Extract on NSAID‐InducedUlcer: An Antioxidative Trail Weaved with Immunomodulatory Effect

Amla (Phyllanthus emblica L.), apart from its food value, can be used as a gastroprotective agent in non steroidal anti-inflammatory drug (NSAID)-induced gastropathy. It has been suggested that the antioxidative property of amla is the key to its therapeutic effect. Hence, on the basis of in vitro antioxidative potential, the ethanolic extract of amla (eAE) was selected for in vivo study in NSAID-induced ulcer. Intriguingly, eAE showed biphasic activity in ulcerated mice, with healing effect observed at 60 mg/kg and an adverse effect at 120 mg/kg.The dose-dependent study revealed that switching from anti-oxidant to pro-oxidant shift and immunomodulatory property could be the major cause for its biphasic effect, as evident from the total antioxidant status, thiol concentration, lipid peroxidation, protein carbonyl content followed by mucin content, PGE2 synthesis and cytokine status. Further, Buthionine sulfoxamine (BSO) pretreatment established the potential impact of antioxidative property in the healing action of eAE. However, eAE efficiently reduced pro-inflammatory cytokine (TNF-α and IL-1β) levels and appreciably upregulate anti-inflammatory cytokine (IL-10) concentration. In conclusion, gastric ulcer healing induced by eAE was driven in a dose-specific manner through the harmonization of the antioxidative property and modulation of anti-inflammatory cytokine level.

Neuroprotective potential of the NF-κB inhibitor peptide IKK-NBD in cerebral ischemia-reperfusion injury

Inflammation plays an important role in ischemic pathology. NF-κB is a transcription factor that has a crucial role in inflammation and cell survival, but its precise role in the ischemic aftermath is still uncertain. Therefore, we evaluated the effect of intracerebroventricular administration of a highly specific NF-κB inhibitor peptide, IKK-NBD, on transient focal cerebral ischemic injury in rat using middle cerebral artery occlusion model. The assessment of ischemia-induced neurological deficits, alterations in the proinflammatory cytokine IL-1β level, OX-42 immunoreactivity, changes in blood–brain barrier (BBB) permeability, reactive oxygen species (ROS) production and DNA fragmentation by terminal dUTP nick end labelling (TUNEL) were monitored at 24 h post reperfusion following 1 h of ischemia after pre-treatment with either 40 μg of IKK-NBD or the inactive IKK-NBD peptide, which served as control. Pre-treatment with IKK-NBD peptide significantly ameliorated the cerebral ischemia-induced neurological deficits. Quantification of IL-1β by ELISA revealed significantly reduced striatal IL-1β level in IKK-NBD peptide treated rats. The treatment also resulted in reduced staining of microglial OX-42 and significantly reduced extravasation of Evans blue dye, indicating protection of BBB from ischemic insult. These results indicate that specific NF-κB inhibition downplays post-ischemic inflammation. Furthermore, reduction in DNA fragmentation as assessed by TUNEL staining also confirms the neuroprotective effect of IKK-NBD peptide. Thus, it may be inferred that IKK-NBD peptide reduces ischemic brain damage and this can, at least partly, be attributed to reduction in inflammation following ischemic injury.

Topical Application of Compound Ibuprofen Suppresses Pain by Inhibiting Sensory Neuron Hyperexcitability and Neuroinflammation in a Rat Model of Intervertebral Foramen Inflammation

There is lack of evidence that topical application of an anti-inflammatory reagent could reduce pain due to intervertebral foramen (IVF) inflammation (IVFI). We investigated analgesic effects and underlying mechanisms of topical application of a compound ibuprofen cream (CIC) onto the surface of back skin covering the inflamed L(5) IVF in a rat model. Repetitive CIC treatment (~.54 g each treatment daily for 5 consecutive days) significantly reduces severity and duration of IVFI-induced thermal hyperalgesia and mechanical allodynia by 80 to 100% and 50 to 66%, respectively. Electrophysiological studies and Western blot analysis demonstrated that CIC treatment significantly inhibited hyperexcitability of the inflamed dorsal root ganglion (DRG) neurons and upregulation of Nav1.7 and Nav1.8 protein, respectively. Pathological manifestations of the inflamed DRG were also markedly improved following CIC treatment. Further, in the inflamed DRGs, phosphorylation and expression of transcription factor NF-κB and pro-inflammatory enzyme cyclooxygenase-2 (COX-2) were significantly increased, while a cytokine IL-1β level was increased. IVFI-induced upregulation of these molecules was significantly inhibited by CIC treatment. This study provides evidence that an anti-inflammatory reagent can be used topically to suppress pain due to IVFI and/or DRG inflammation through inhibition of sensory neuron hyperexcitability and the immune and inflammatory responses. This study suggests a convenient and safe clinical intervention for treating pain due to intervertebral foramen inflammation and similar syndromes.

Protective effects of indomethacin-loaded nanocapsules against oxygen-glucose deprivation in organotypic hippocampal slice cultures: Involvement of neuroinflammation

Targeted treatment of diseases of the central nervous system remains problematic due to the complex pathogenesis of these disorders and the difficulty in drug delivery. Here we investigate the neuroprotective effect of indomethacin-loaded nanocapsules (IndOH-NC) in an in vitro model of ischemia. For this purpose we used organotypic hippocampal cultures exposed to oxygen-glucose deprivation (OGD). When the cultures were exposed to 60 min of OGD, 54.5 ± 14.7% of the total area of the hippocampal slices was labeled with propidium iodide. On the other hand, when the cultures were treated with 50 or 100 μM of IndOH-NC the cell death was significantly reduced to 31 ± 7% ( P < 0.05) and 20 ± 4% ( P < 0.001), respectively. The treatment with IndOH-NC markedly inhibited the levels of the pro-inflammatory cytokines IL-1β, IL-6 and TNF-α levels even 48 h after OGD. Immunoblotting revealed that treatment with 100 μM of IndOH-NC was able to significantly reduce to the levels of control cultures the levels of ERK1/2 and JNK phosphorylation, as well as iNOS activation. Additionally, IndOH-NC prevented glial activation induced by OGD, as evidenced by a decrease of GFAP immunocontent and Isolectin B 4 reactivity. Our results clearly demonstrate that IndOH-NC might represent a promising pharmaceutical neuroprotective formulation for cerebral ischemia, most probably by inhibiting the inflammatory cascades.

Distinct Bifidobacterium strains drive different immune responses in vitro

In this work we evaluated the specific immune activation properties of different Bifidobacterium strains, some of the most relevant intestinal microorganisms. To this end, we examined the in vitro effect of 12 Bifidobacterium strains belonging to 4 different species, Bifidobacterium longum, Bifidobacterium breve, Bifidobacterium bifidum and Bifidobacterium animalis subsp. lactis, on the maturation pattern of human monocyte-derived dendritic cells (DCs), as well as in their ability to induce cytokine secretion. In addition, we determined peripheral blood mononuclear cell (PBMC) proliferation and cytokine expression after exposure to bacterial strains. All bifidobacteria tested were able to induce full DC maturation but showed differences in the levels of cytokine production, especially IL-12, IL-10, TNFα and IL-1β, suggesting that specific cytokine ratios could be used to predict the type of Th response that they may promote. In fact, analysis of cytokine production by PBMC showed that most of the tested B. animalis and B. longum strains induced the secretion of large amounts of IFNγ and TNFα, in agreement with the Th1 profile suggested by DC cytokine production. Remarkably, three of four B. bifidum strains induced poor secretion of these cytokines and significant amounts of IL-17, the main product of Th17 cells, in accordance with the high IL-1β/IL-12 ratio observed after DC stimulation. In conclusion, this work shows species and strain-specific immune effects of bifidobacteria and describes a valuable method for screening possible probiotic strains with different immunomodulatory properties. Notably, some B. bifidum strains seem to promote Th17 polarization, which could be useful for future probiotic applications.

Role of galectin-3 in macrophage response to Listeria monocytogenes infection (133.34)

Abstract Galectin-3 is a β-galactoside-binding animal lectin expressed in various immune cell types including macrophages. It has been implicated as a positive regulator of macrophage phagocytosis and as a negative regulator of LPS-mediated inflammation. However, its role in macrophage antibacterial function has not been fully elucidated. The aim of this study was to investigate the role of galectin-3 in macrophage antibacterial function against Listeria monocytogenes (LM) infection by studying bone marrow-derived macrophages (BMM) from galectin-3-deficient (gal3-/-) and wild-type (gal3+/+) mice. After in vitro infection, gal3-/- BMM contained significantly fewer viable intracellular bacteria than gal3+/+ BMM. Consistently, immunofluorescent staining of infected BMM revealed a significantly lower percentage of cytosolic LM escaped from phagosomes in gal3-/- BMM compared to gal3+/+ BMM. Higher levels of reactive nitrogen intermediate nitric oxide (NO) but lower levels of inflammatory cytokine IL-1β were detected in LM-infected gal3-/- BMM compared to gal3+/+ cells. In addition, we noted elevated levels of NAD(P)H in gal3-/- BMM which may contribute to higher amounts of NO in gal3-/- BMM. We conclude that galectin-3 plays a regulatory role in macrophage antibacterial function by interfering with the production of NO, thereby facilitating the survival and replication of intracellular bacteria.

Interleukin-1β, Interleukin-18, and Interferon-γ Expression in the Cerebrospinal Fluid of Premature Infants with Posthemorrhagic Hydrocephalus—Markers of White Matter Damage?

Posthemorrhagic hydrocephalus (PHHC) represents a major complication of preterm birth. The aim of this study was to determine whether cerebrospinal fluid (CSF) levels of the pro-inflammatory cytokines IL-1beta, IL-18, and interferon (IFN)-gamma are altered in the CSF of preterm infants with PHHC and may serve as a marker of white matter damage (WMD). Twenty-seven preterm infants with PHHC were included in the study; 13 of them had signs of cystic WMD (cWMD) on ultrasound examinations. CSF sample 1 was obtained at first ventriculostomy, sample 2 at shunt implantation. Results were compared with a control group of 20 age-matched patients without neurologic diseases. IL-1beta concentrations were elevated in CSF sample 1 of PHHC patients without WMD and in sample 1 of patients with cWMD. Concentrations of IL-18 were increased in both samples of patients without WMD and in sample 2 of patients with cWMD. CSF levels of IFN-gamma were elevated in sample 1 of PHHC patients with cWMD. The pro-inflammatory cytokine IL-1beta and IL-18 levels in the CSF are elevated in patients with PHHC. Higher IFN-gamma levels are detected in a subgroup of patients developing cWMD, indicating its involvement in the pathogenesis of cWMD in the context of PHHC.

Suppression of endothelial or lipoprotein lipase in THP-1 macrophages attenuates proinflammatory cytokine secretion

LPL and endothelial lipase (EL) are associated with macrophages in human atherosclerotic lesions, and overexpression of LPL in mouse macrophages is associated with a greater extent of atherosclerosis. To investigate potential mechanisms by which macrophage-derived lipase expression may mediate proatherogenic effects, we used lentivirus-mediated RNA interference to suppress the expression of either LPL or EL within THP-1 macrophages. After suppression of either LPL or EL, significant decreases in the concentration of interleukin-1beta, interleukin-6, monocyte chemoattractant protein-1, and tumor necrosis factor-alpha were observed. Incubation of THP-1 macrophages with either mildly or extensively oxidized LDL consistently decreased cytokine expression, which was additive to that contributed by lipase suppression. Decreased lipase expression was also associated with an altered lipid composition, with reduced percentages of cholesterol (unesterified and esterified), triglycerides, and lysophosphatidylcholine. Microarray data indicated a decreased expression of proinflammatory genes, growth factors, and antiapoptotic genes. By contrast, there was an increased expression of lipoprotein receptors (scavenger receptor 1, low density lipoprotein receptor, scavenger receptor class B type I, and CD36). Thus, we conclude that the suppression of either LPL or EL decreases proinflammatory cytokine expression and influences the lipid composition of THP-1 macrophages. These results provide further insight into the specific metabolic and potential pathological roles of LPL and EL in human macrophages.

Pioglitazone, a peroxisome proliferator-activated receptor γ activator, ameliorates experimental autoimmune myocarditis by modulating Th1/Th2 balance

Objectives – The aim of this study is to investigate the effect of Peroxisome proliferator-activated receptor γ (PPARγ) ligand on experimental autoimmune myocarditis (EAM). Background – Rat EAM model resembles the giant cell myocarditis in human. Recent studies have suggested that Th1 cytokines are involved in the initiation and progression of EAM, whereas Th2 cytokines are associated with the remission. PPARγ, which is a member of nuclear hormone receptor superfamily, has been known to affect not only glucose homeostasis but also immune responses, by regulating the Th1/Th2 balance. Methods – Lewis rats were immunized with cardiac myosin and divided into three groups: Group N, normal control rats ( n = 16); Group E, EAM rats ( n = 17); and Group P, EAM rats treated with a PPARγ activator pioglitazone ( n = 20). Results – Cardiac dysfunction and remodeling were inhibited in Group P compared to Group E. Heart weight/body weight ratio and the degree of inflammation and fibrosis were significantly lower in Group P than in Group E. The mRNA levels of macrophage inflammatory protein-1α (MIP-1α), which plays an important role in the recruitment of inflammatory cells in the early stage of EAM, were upregulated in the heart of Group E, but not in the heart of Group P. Furthermore, the treatment with pioglitazone decreased the expression levels of proinflamatory cytokine (TNFα and IL-1β) genes and Th1 cytokine (IFN-γ) genes, and increased the expression levels of Th2 cytokine (IL-4) gene. Conclusions – PPARγ ligands may have beneficial effects on myocarditis by inhibiting MIP-1α expression and modulating the Th1/Th2 balance.

Water Extracts of Helicobacter pylori Delay Healing of Chronic Gastric Ulcers in Rats: Role of Cytokines and Gastrin-Somatostatin Link

Background: Helicobacter pylori (Hp) is considered as a major risk factor of peptic ulcer, but the pathogenic mechanism of its action has not been fully explained. Aims: This study was designed: (1) to compare the ulcer healing effects of water extract (WE) obtained from type-I cytotoxin-associated gene A (CagA) and vacuolating cytotoxin A (VacA) expressing Hp and from type-II CagA- and VacA-negative Hp strain with those of vehicle (saline), and (2) to determine the alterations in gastric secretion, gastric blood flow (GBF) and expression of Hp-related cytokines during the ulcer healing in rats treated with toxigenic (type-I) and non-toxigenic (type-II) Hp-derived WE. Methods: Gastric ulcers were produced by serosal application of acetic acid in rats with or without gastric fistula treated with vehicle (saline) or WE originating from type-I or type-II Hp administered intragastrically on days 1, 3, 5 and 7 upon ulcer induction. On days 3, 9 and 15, animals were lightly anesthetized with ether, the abdomen was opened and the GBF was measured by the H₂-gas clearance technique in the ulcer area and non-ulcerated mucosa. Venous blood was withdrawn for the measurement of plasma cytokine (IL-1β and TNFα) levels and plasma and gastric contents were also collected for gastrin and somatostatin determination by specific radioimmunoassay. Results: Gastric ulcers healed gradually in vehicle-treated controls and the ulcer area on days 3, 9 and 15 was reduced by 12, 43 and 92%, respectively. In rats treated with WE of type-I Hp, ulcer healing was significantly delayed, and gastritis and infiltration of ulcerated gastric mucosa with inflammatory cells were observed histologically. The prolongation of ulcer healing by WE of both Hp strains was accompanied by a marked fall in the GBF at the ulcer margin and transient hyposecretion especially in rats given WE of type-I Hp strain. On day 15 of ulcer healing, the plasma concentration of IL-1β and TNFα was negligible in vehicle control rats, but it was significantly elevated particularly in rats treated with WE of type-I Hp. RT-PCR analysis revealed that mucosal expression of IL-1β and TNFα mRNA was significantly upregulated in the gastric mucosa of rats treated with either toxigenic or non-toxigenic Hp WE. The plasma gastrin level was significantly higher and the luminal concentration of somatostatin was significantly lower in rats treated with Hp-WE than in vehicle-treated controls and these alterations were more pronounced in rats treated with WE type-I than type-II Hp. Conclusions: WE of toxigenic Hp strain delays ulcer healing due to the reduction in the gastric microcirculation at the ulcer margin, the overexpression of inflammatory cytokines and the impairment of the gastrin-somatostatin link.

Monoclonal antibody based enzyme-linked and chemiluminescent assays for the human interleukin-1 receptor antagonist Application to measure hIL-1ra levels in monocyte cultures and synovial fluids

Interleukin-1 receptor antagonist (IL-1ra) has the potential to counteract at least part of the biological effects of interleukin-1. The outcome of an inflammatory reaction may therefore be determined by the balance between IL-1 and IL-1ra, rather than by IL-1 alone. We have developed an immunoassay to address this issue as well as to assess the effects of anti-inflammatory agents on the expression of IL-1 and IL-1ra in vitro or in body fluids. Recombinant human IL-1ra was expressed in an E. coli system, purified to homogeneity, and used to derive monoclonal antibodies in mice as well as polyclonal antibodies in rabbits. A sandwich ELISA was constructed with F(ab′) 2 fragments of a high affinity monoclonal antibody and the rabbit serum as a source of secondary antibody. The assay required no sample treatment to avoid interference by rheumatoid factor. The measuring range was 0.020-2 ng/ml. By labelling a second monoclonal antibody with an acridinium ester, a chemiluminescence assay with a wider measuring range (0.050–15 ng/ml) was generated. In accord with published data, we found that IL-1ra was secreted by human monocytes stimulated with LPS, Zymosan, IL-1α, or human IgG. After an induction phase of ca. 4 hours and depending on the stimulus, IL-1ra accumulated linearly for periods up to 96 h. IL-1ra levels in synovial fluids of 19 patients suffering from various inflammatory joint diseases were compared with the cytokine levels of IL-1β, IL-g, IL-8, and TNF-α. Highest positive correlations were found with IL-8 and IL-1β. In normal blood donors IL-1ra serum levels were 150–800 pg/ml (Median: 387 pg/ml). Owing to its sensitivity and large measuring range the newly developed assays appear to be suitable for measuring IL-1ra in cell cultures as well as in biological fluids.