interleukin-8 and 4-hydroxy-2-nonenal

Endometriosis is a common gynaecological disease of women in reproductive age, and is thought to arise from retrograde menstruation and implantation of endometrial tissue, mostly into the peritoneal cavity. The condition is characterized by a chronic, unresolved inflammatory process thereby contributing to pain as cardinal symptom in endometriosis. Elevated reactive oxygen species (ROS) and oxidative stress have been postulated as factors in endometriosis pathogenesis. We here set out for a systematic study to identify novel mechanisms and pathways relating to oxidative stress in ectopic peritoneal lesions. Using combined proteomic and transcriptomic approaches, we identified novel targets including upregulated pro-oxidative enzymes, such as amine oxidase 3/vascular adhesion protein 1 (AOC3/VAP1) as well as downregulated protective factors, in particular alkenal reductase PTGR1 and methionine sulfoxide reductase. Consistent with an altered ROS landscape, we observed hemoglobin / iron overload, ROS production and lipid peroxidation in ectopic lesions. ROS-derived 4-hydroxy-2-nonenal induced interleukin IL-8 release from monocytes. Notably, AOC3 inhibitors provoked analgesic effects in inflammatory pain models in vivo, suggesting potential translational applicability.

Topics: Aldehydes; Allyl Compounds; Amine Oxidase (Copper-Containing); Analgesics; Animals; Biomarkers; Cell Adhesion Molecules; Disease Models, Animal; Endometriosis; Female; Gene Expression Profiling; Heme; Humans; Inflammation Mediators; Interleukin-8; Iron; Lipid Peroxidation; Metabolic Networks and Pathways; Mice; Mice, Inbred BALB C; Myeloid Cells; Oxidative Stress; Peritoneal Diseases; Phagocytosis; Sulfonamides

4-Hydroxynonenal (HNE) is a highly reactive end-product of lipid peroxidation reaction that leads to retinal pigment epithelial (RPE) cell damage. Cyanidin-3-glucoside (C3G), the most abundant anthocyanin in the edible parts of plants, is a nutritional supplement used for preventing retinal damage. However, the protective effect of C3G against HNE-induced RPE cell damage remains to be elucidated. The protective mechanisms of C3G on ARPE-19 cells after HNE exposure were investigated in this study. Results showed that compared with HNE-treated cells, the viability of ARPE-19 cells was significantly (P < 0.05) increased after 1 and 5 μM C3G treatment. C3G exhibited a significant (P < 0.05) inhibitory effect on the expression of senescence-associated β-galactosidase in ARPE-19 cells. VEGF levels in the C3G groups were significantly (P < 0.05) decreased relative to those of the HNE-treated group. C3G also regulated the release of two inflammatory mediators, namely monocyte chemoattractant protein 1 and interleukine-8, in ARPE-19 cells after HNE treatment. Furthermore, C3G attenuated retinal cell apoptosis in pigmented rabbits induced by visible light. Therefore, our data showed that C3G has efficient protective effects on HNE-induced apoptosis, angiogenesis, and dysregulated cytokine production in ARPE-19 cells.

Topics: Aldehydes; Animals; Anthocyanins; Apoptosis; Chemokine CCL2; Glucosides; Humans; Interleukin-8; Light; Rabbits; Retinal Diseases; Retinal Pigment Epithelium; Vascular Endothelial Growth Factor A

Recent findings support a connection between mitochondrial dysfunction and activation of inflammatory pathways in articular cells. This study investigates in vivo in an acute model whether intra-articular administration of oligomycin, an inhibitor of mitochondrial function, induces an oxidative and inflammatory response in rat knee joints.. Oligomycin was injected into the rat left knee joint on days 0, 2, and 5 before joint tissues were obtained on day 6. The right knee joint served as control. Results were evaluated by macroscopy and histopathology and by measuring cellular and mitochondrial reactive oxygen species (ROS), 4-hydroxy-2-nonenal (4-HNE, a marker of lipid peroxidation), nuclear factor erythroid 2-related factor 2 (Nrf2), and CD68 (macrophages) and chemokine levels. The marker of mitochondrial mass COX-IV was also evaluated.. The macroscopic findings showed significantly greater swelling in oligomycin-injected knees than in control knees. Likewise, the histological score of synovial damage was also increased significantly. Immunohistochemical studies showed high expression of IL-8, coinciding with a marked infiltration of polymorphonuclears and CD68+ cells in the synovium. Mitochondrial mass was increased in the synovium of oligomycin-injected joints, as well as cellular and mitochondrial ROS production, and 4-HNE. Relatedly, expression of the oxidative stress-related transcription factor Nrf2 was also increased. As expected, no histological differences were observed in the cartilage; however, cytokine-induced neutrophil chemoattractant-1 mRNA and protein expression were up-regulated in this tissue.. Mitochondrial failure in the joint is able to reproduce the oxidative and inflammatory status observed in arthritic joints.

Topics: Aged; Aged, 80 and over; Aldehydes; Animals; Antigens, CD; Antigens, Differentiation, Myelomonocytic; Arthritis, Experimental; Cartilage, Articular; Chemokine CXCL1; Electron Transport Complex IV; Enzyme Inhibitors; Female; Fluorescent Antibody Technique; Humans; Immunohistochemistry; Injections, Intra-Articular; Interleukin-8; Knee Joint; Macrophages; Middle Aged; Mitochondria; Mitochondrial Proton-Translocating ATPases; NF-E2-Related Factor 2; Oligomycins; Osteoarthritis; Rats; Rats, Wistar; Reactive Oxygen Species; Synovial Membrane

Cigarette smoke (CS) is a main source of oxidative stress and a key risk factor for emphysema, which consists of alveolar wall destruction. Alveolar type (AT) II cells are in the gas exchange regions of the lung. We isolated primary ATII cells from deidentified organ donors whose lungs were not suitable for transplantation. We analyzed the cell injury obtained from nonsmokers, moderate smokers, and heavy smokers. DJ-1 protects cells from oxidative stress and induces nuclear erythroid 2-related factor-2 (Nrf2) expression, which activates the antioxidant defense system. In ATII cells isolated from moderate smokers, we found DJ-1 expression by RT-PCR, and Nrf2 and heme oxygenase (HO)-1 translocation by Western blotting and immunocytofluorescence. In ATII cells isolated from heavy smokers, we detected Nrf2 and HO-1 cytoplasmic localization. Moreover, we found high oxidative stress, as detected by 4-hydroxynonenal (4-HNE) (immunoblotting), inflammation by IL-8 and IL-6 levels by ELISA, and apoptosis by terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) assay in ATII cells obtained from heavy smokers. Furthermore, we detected early DJ-1 and late Nrf2 expression after ATII cell treatment with CS extract. We also overexpressed DJ-1 by adenovirus construct and found that this restored Nrf2 and HO-1 expression and induced nuclear translocation in heavy smokers. Moreover, DJ-1 overexpression also decreased ATII cell apoptosis caused by CS extract in vitro. Our results indicate that DJ-1 activates the Nrf2-mediated antioxidant defense system. Furthermore, DJ-1 overexpression can restore the impaired Nrf2 pathway, leading to ATII cell protection in heavy smokers. This suggests a potential therapeutic strategy for targeting DJ-1 in CS-related lung diseases.

Topics: Adenoviridae; Aldehydes; Alveolar Epithelial Cells; Apoptosis; Cell Separation; Cytoprotection; Humans; Interleukin-6; Interleukin-8; NF-E2-Related Factor 2; Protein Deglycase DJ-1; RNA, Messenger; Smoking

It is now thought that atherosclerosis, although due to increased plasma lipids, is mainly the consequence of a complicated inflammatory process, with immune responses at the different stages of plaque development. Increasing evidence points to a significant role of Toll-like receptor 4 (TLR4), a key player in innate immunity, in the pathogenesis of atherosclerosis. This study aimed to determine the effects on TLR4 activation of two reactive oxidized lipids carried by oxidized low-density lipoproteins, the oxysterol 27-hydroxycholesterol (27-OH) and the aldehyde 4-hydroxynonenal (HNE), both of which accumulate in atherosclerotic plaques and play a key role in the pathogenesis of atherosclerosis. Secondarily, it examined their potential involvement in mediating inflammation and extracellular matrix degradation, the hallmarks of high-risk atherosclerotic unstable plaques. In human promonocytic U937 cells, both 27-OH and HNE were found to enhance cell release of IL-8, IL-1β, and TNF-α and to upregulate matrix metalloproteinase-9 (MMP-9) via TLR4/NF-κB-dependent pathway; these actions may sustain the inflammatory response and matrix degradation that lead to atherosclerotic plaque instability and to their rupture. Using specific antibodies, it was also demonstrated that these inflammatory cytokines increase MMP-9 upregulation, thus enhancing the release of this matrix-degrading enzyme by macrophage cells and contributing to plaque instability. These innovative results suggest that, by accumulating in atherosclerotic plaques, the two oxidized lipids may contribute to plaque instability and rupture. They appear to do so by sustaining the release of inflammatory molecules and MMP-9 by inflammatory and immune cells, for example, macrophages, through activation of TLR4 and its NF-κB downstream signaling.

Topics: Aldehydes; Cell Line; Gene Expression Regulation; Humans; Hydroxycholesterols; Interleukin-1beta; Interleukin-8; Matrix Metalloproteinase 9; Models, Biological; Monocytes; NF-kappa B; Plaque, Atherosclerotic; RNA, Small Interfering; Signal Transduction; Toll-Like Receptor 4; Tumor Necrosis Factor-alpha

Topics: Aging; Aldehydes; Animals; Biomarkers; Heme Oxygenase-1; Inflammation; Interleukin-6; Interleukin-8; Mice; Mice, Inbred Strains; Models, Animal; NADPH Oxidases; Oxidative Stress; Ozone; Skin; Smoking

Bisphenol A (BPA) is an endocrine-disrupting chemical that leaches from polycarbonate plastics that consequently leads to low-dose human exposure. In addition to its known xenoendocrine action, BPA exerts a wide variety of metabolic effects, but no data are available on its actions on the functions of liver mitochondrial. To assess these effects, HepG2 cells were exposed to BPA (10(-4)-10(-12)M) and physiological parameters were measured by flow cytometry. We demonstrated a significant mitochondrial dysfunction including ROS production, ΔΨ(M) hyperpolarization, lipid accumulation, lipoperoxidation and the release of pro-inflammatory cytokines. In conclusion, we showed that low concentrations of BPA promote lipid accumulation in hepatic cells triggered by disturbances in mitochondrial function, alterations in lipid metabolism and by inflammation that can therefore contribute to steatosis.

Topics: Aldehydes; Benzhydryl Compounds; Endocrine Disruptors; Hep G2 Cells; Humans; Interleukin-8; Lipid Metabolism; Lipid Peroxidation; Membrane Potential, Mitochondrial; Mitochondria; Nitric Oxide; Phenols; Reactive Oxygen Species; Tumor Necrosis Factor-alpha

Human esophageal epithelium is continuously exposed to physical stimuli or to gastric acid that sometimes causes inflammation of the mucosa. Transient receptor potential vanilloid 1 (TRPV1) is a nociceptive, Ca(2+)-selective ion channel activated by capsaicin, heat, and protons. It has been reported that activation of TRPV1 expressed in esophageal mucosa is involved in gastroesophageal reflux disease (GERD) or in nonerosive GERD symptoms. In this study, we examined the expression and function of TRPV1 in the human esophageal epithelial cell line Het1A, focusing in particular on the role of oxidative stress. Interleukin-8 (IL-8) secreted by Het1A cells upon stimulation by capsaicin or acid with/without 4-hydroxy-2-nonenal (HNE) was measured by ELISA. Following capsaicin stimulation, the intracellular production of reactive oxygen species (ROS) was determined using a redox-sensitive fluorogenic probe, and ROS- and HNE-modified proteins were determined by Western blotting using biotinylated cysteine and anti-HNE antibody, respectively. HNE modification of TRPV1 proteins was further investigated by immunoprecipitation after treatment with synthetic HNE. Capsaicin and acid induced IL-8 production in Het1A cells, and this production was diminished by antagonists of TRPV1. Capsaicin also significantly increased the production of intracellular ROS and ROS- or HNE-modified proteins in Het1A cells. Moreover, IL-8 production in capsaicin-stimulated Het1A cells was enhanced by synthetic HNE treatment. Immunoprecipitation studies revealed that TRPV1 was modified by HNE in synthetic HNE-stimulated Het1A cells. We concluded that TRPV1 functions in chemokine production in esophageal epithelial cells, and this function may be regulated by ROS via posttranslational modification of TRPV1.

Topics: Acids; Aldehydes; Animals; Calcium Channels; Capsaicin; Cell Line; Epithelial Cells; Esophagus; Humans; Interleukin-8; Male; Mucous Membrane; Oxidative Stress; Rats; Rats, Wistar; Reactive Oxygen Species; TRPV Cation Channels

The transient receptor potential subfamily A member 1 (TRPA1) is a non-selective cation channel implicated in the pathogenesis of several airway diseases like asthma and chronic obstructive pulmonary disease (COPD). Most of the research on TRPA1 focuses on its expression and function in neuronal context; studies investigating non-neuronal expression of TRPA1 are lacking. In the present study, we show functional expression of TRPA1 in human lung fibroblast cells (CCD19-Lu) and human pulmonary alveolar epithelial cell line (A549). We demonstrate TRPA1 expression at both mRNA and protein levels in these cell types. TRPA1 selective agonists like allyl isothiocyanate (AITC), 4-hydroxynonenal (4-HNE), crotonaldehyde and zinc, induced a concentration-dependent increase in Ca+2 influx in CCD19-Lu and A549 cells. AITC-induced Ca+2 influx was inhibited by Ruthenium red (RR), a TRP channel pore blocker, and by GRC 17536, a TRPA1 specific antagonist. Furthermore, we also provide evidence that activation of the TRPA1 receptor by TRPA1 selective agonists promotes release of the chemokine IL-8 in CCD19-Lu and A549 cells. The IL-8 release in response to TRPA1 agonists was attenuated by TRPA1 selective antagonists. In conclusion, we demonstrate here for the first time that TRPA1 is functionally expressed in cultured human lung fibroblast cells (CCD19-Lu) and human alveolar epithelial cell line (A549) and may have a potential role in modulating release of this important chemokine in inflamed airways.

Topics: Aldehydes; Calcium; Calcium Channels; Cations, Divalent; Cells, Cultured; Chlorides; Dose-Response Relationship, Drug; Epithelial Cells; Fibroblasts; Humans; Interleukin-8; Isothiocyanates; Nerve Tissue Proteins; Ruthenium Red; Transient Receptor Potential Channels; TRPA1 Cation Channel; Zinc Compounds

This study was to explore the role of erythromycin on the synthesis of interleukin-8 (IL-8) and gamma-GCS treated by 4-hydroxynonenal (4-HNE) in bronchial epithelial cells (16-HBE).. (1) The experiment groups were divided into 10 micromol/L 4-HNE and control groups. The phosphorylation of ERK-1, JNK, P38MAPK and the combining activity of AP-1 after 10 micromol/L 4-HNE stimulating for 0.5, 2, 4, 8, 12 hours were all estimated. (2) The IL-8 and IL-8 mRNA, gamma-GCS and gamma-GCS mRNA were measured after 4-HNE (or serum-free medium) stimulating 0.5, 2, 4, 8, 12 hours. (3) The effects of PD98059 and erythromycin on the expression of gamma-GCS, gamma-GCS mRNA, IL-8 and erythromycin on AP-1 combining activity by the 4-HNE were all detected.. (1) The level of phosphorylation of ERK1 in the 4-HNE and control groups were 110.4+/-1.6, 114.6+/-2.4, 106.1+/-2.1, 110.2+/-2.0, 104.4+/-3.4, 112.8+/-2.4, 96.3+/-9.6, 115.4+/-3.8, 86.3+/-2.8, 113.1+/-2.6 at 0.5, 2, 4, 8, 12 hours, respectively, P<0.05. While the AP-1 combining activity in the 4-HNE and control groups were 90.6+/-2.0, 98.6+/-2.1, 85.7+/-2.2, 98.7+/-3.4, 78.2+/-2.6, 100.1+/-3.8, 70.6+/-1.8, 101.3+/-4.2, 64.9+/-4.8, 97.4+/-3.6, respectively P<0.05. (2) The level of IL-8 in the 4-HNE and control groups at 2, 4, 8, 12 hours were (87.4+/-3.8) microg/L, (63.9+/-3.8) microg/L, (98.3+/-4.2) microg/L, (65.3+/-6.2) microg/L, (102.4+/-5.7) microg/L, (64.6+/-4.8) microg/L, (116.5+/-5.6) microg/L, (63.7+/-6.6) microg/L, respectively, P<0.05. The levels of gamma-GCS at 2, 4, 8, 12 hours in two groups were 5.43+/-0.23, 4.78+/-0.26, 5.41+/-0.27, 4.03+/-0.34, 5.54+/-0.53, 3.22+/-0.31, respectively, P<0.05. IL-8 mRNA, gamma-GCS mRNA after 4-HNE stimulation were all increased compared with the control groups. (3) The expression of IL-8 and AP-1 combining activity was decreased, but synthesis of gamma-GCS was increased after treatment with PD98059 or erythromycin before treated with 4-HNE.. 4-HNE could increase the expression of IL-8 in the bronchial epithelial cells, via increasing the transcribing activities of AP-1 via ERK-1 cell signal transduction ways. Erythromycin could inhibit the synthesis of IL-8 by blocking AP-1 pathway. PD98059 and erythromycin could block AP-1 transduction pathway, but increase the synthesis of gamma-GCS induced by 4-HNE in bronchial epithelial cells.

Topics: Aldehydes; Bronchi; Cells, Cultured; Epithelial Cells; Erythromycin; Flavonoids; Glutamate-Cysteine Ligase; Humans; Interleukin-8; Transcription Factor AP-1

4-Hydroxynonenal (4-HNE) can increase the synthesis of interleukin-8 (IL-8) in bronchial epithelium cells (16HBE). This study was to explore the role of ginkgolide B in inhibiting the synthesis of IL-8 induced by 4-HNE in 16HBE.. The experiments were divided into 3 groups: a group treated with 4-HNE (10 micromol/L), a group treated with ginkgolide B (100 micromol/L) + 4-HNE (10 micromol/L), and a control group. IL-8 and IL-8 mRNA were measured after 4-HNE (or serum-free medium) stimulation for 0.5, 2, 4, 8, 12 hours. The phosphorylation of ERK1/2, JNK, p38MAPK and the combining activity of AP-1 after 4-HNE stimulation for 0.5, 2, 4, 8, 12 hours were all examined. IL-8 and the combining activity of AP-1 were measured after the 16HBE were pre-incubated with 50 micromol/L PD98059 (MEK1 inhibitor) for 2 hours before 4-HNE stimulation. The combining activity of AP-1 in 10 micromol/L 4-HNE, 100 micromol/L ginkgolide B + 10 micromol/L 4-HNE, and the control groups were all measured by EMSA.. The level of IL-8 in 10 micromol/L 4-HNE, 100 micromol/L ginkgolide B + 10 micromol/L 4-HNE, the control groups after 4-HNE stimulating for 4 h were (98.3 +/- 4.2), (88.2 +/- 5.3), (65.3 +/- 6. 2) and (116.5 +/- 5.6), (102.8 +/- 4.7), (63.7 +/- 6.6) microg/L for 12 h. The level of IL-8 and IL-8 mRNA after 4-HNE stimulation in the ginkgolide B + 4-HNE group were lower than those in the 4-HNE group while higher than those in the control groups. The level of phosphorylation of ERK1 in the 4-HNE group at 0.5, 2, 4, 8, 12 hours were higher than those in the control groups (t = 2.83 - 14.03, P < 0.05). The AP-1 combining activity in the 4-HNE group, the ginkgolide B + 4-HNE group, PD98059 + 4-HNE group, and the control group were significantly different (F = 21.49 - 194.16, P < 0.01). The expression of IL-8 and the AP-1 combining activity in groups of pre-incubated with PD98059 2 hours before 4-HNE stimulation were lower than that without PD98059. The combining activity of AP-1 in the ginkgolide B + 4-HNE group was decreased as compared to the 4-HNE groups.. 4-HNE increased the expression of Interleukin-8 in bronchial epithelium cells, via increasing the transcription activities of AP-1 by ERK1 cell signal transduction pathways. Ginkgolide B inhibited synthesis of IL-8 by blocking ERK1-AP1 transduction pathways.

Topics: Aldehydes; Cells, Cultured; Drug Antagonism; Epithelial Cells; Ginkgolides; Humans; Interleukin-8; Lactones; MAP Kinase Signaling System; Proto-Oncogene Proteins c-jun

Oxidized low density lipoproteins (oxLDLs) may exert several pro-inflammatory effects that can contribute to the development of coronary artery disease (CAD). Evaluating a possible correlation between oxLDLs and clinical expression of CAD, we measured specific lipid peroxidation indices in healthy subjects and in patients at different clinical stages of CAD. We observed a slight, but not significant, increase in plasma content of cholesterol oxidation products, i.e. oxysterols, in all CAD patients, and a slight, but not significant, increase of 4-hydroxynonenal-protein adducts only in subjects with acute CAD. Moreover, CAD patients showed a plasma rise of specific inflammatory proteins, i.e. C-reactive protein, intercellular adhesion molecule-1, and interleukin-8, but not of monocyte chemotactic protein-1. These preliminary data, without excluding an involvement of oxidative stress and inflammation in CAD, do not show a strict correlation between relevant plasma markers, other than C-reactive protein, and acute phase of the disease.

Topics: Aged; Aged, 80 and over; Aldehydes; Antigens, CD; Biomarkers; Blood Proteins; C-Reactive Protein; Cell Adhesion Molecules; Chemokine CCL2; Coronary Disease; Female; Humans; Inflammation; Interleukin-8; Lipid Peroxidation; Male; Middle Aged; Reference Values

Lipolytic products of triglyceride-rich lipoproteins, i.e., free fatty acids, may cause activation and dysfunction of the vascular endothelium. Mechanisms of these effects may include lipid peroxidation. One of the major and biologically active products of peroxidation of n-6 fatty acids, such as linoleic acid or arachidonic acid, is the aldehyde 4-hydroxynonenal (HNE). To study the hypothesis that HNE may be a critical factor in endothelial cell dysfunction caused by free fatty acids, human umbilical endothelial cells (HUVEC) were treated with up to160 microM of linoleic or arachidonic acid. HNE formation was detected by immunocytochemistry in cells treated for 24 h with either fatty acid, but more markedly with arachidonic acid. To study the cellulareffects of HNE, HUVEC were treated with different concentrations of this aldehyde, and several markers of endothelial cell dysfunction were determined. Exposure to HNE for 6 and 9 h resulted in increased cellular oxidative stress. However, short time treatment with HNE did not cause activation of nuclear factor-kappaB (NF-kappaB). In addition, HUVEC exposure to HNE caused a dose-dependent decrease in production of both interleukin-8 (IL-8) and intercellular adhesion molecule-1 (ICAM-1). On the other hand, HNE exerted prominent cytotoxic effects in cultured HUVEC, manifested by morphological changes, diminished cellular viability, and impaired endothelial barrier function. Furthermore, HNE treatment induced apoptosis of HUVEC. These data provide evidence that HNE does not contribute to NF-kappaB-related mechanisms of the inflammatory response in HUVEC, but rather to endothelial dysfunction, cytotoxicity, and apoptotic cell death.

Topics: Aldehydes; Animals; Apoptosis; Cell Survival; Cells, Cultured; Endothelium, Vascular; Gene Expression Regulation; Humans; Immunohistochemistry; Intercellular Adhesion Molecule-1; Interleukin-8; Kinetics; Pulmonary Artery; Swine; Time Factors; Umbilical Veins