1-3-dihydroxy-4-4-5-5-tetramethyl-2-(4-carboxyphenyl)tetrahydroimidazole and Inflammation

Radiation-induced bystander effects are a well-known phenomenon that are observed when treating cancer and other diseases after radiotherapy, and even after occupational exposure to radiation. However, little is known about the crosstalk between irradiated macrophages and endothelial cells that line the circulatory system, which may play a role in the development of atherosclerosis. In the current study, we found that the expression of inducible nitric oxide synthase (iNOS) and the intracellular level of nitric oxide (NO) in gamma-irradiated U937 macrophage cells were significantly increased. When human umbilical vein endothelial cells (HUVECs) were co-cultured with gamma-irradiated U937 cells, additional micronuclei (MN) and apoptosis were induced so that the plating efficiency of the bystander HUVECs decreased and P38 was overexpressed in the bystander HUVECs cells. In addition, the contents of vascular cell adhesion molecule 1 (VCAM-1) and the activities of matrix metalloproteinase-9 (MMP-9) in the culture medium of bystander HUVECs were increased. Furthermore, during cell co-culture the adhesive ability of irradiated U937 cells to the bystander HUVECs increased. When U937 cells were treated with 500 μM S-methylisothiourea sulfate (SMT) (iNOS inhibitor) before irradiation, and HUVECs were treated with 10 μM SB203580 (p38 inhibitor) before cell co-culture or treated with 20 μM c-PTIO (NO scavenger) in the co-culture medium, the bystander micronuclei and the amounts of VCAM-1 and MMP-9 in the medium of bystander HUVECs were diminished, and the ability of irradiated U937 cells adhering to HUVECs was also reduced, while the plating efficiency of bystander HUVECs partially recovered. These results demonstrated that irradiated U937 cells appear to release nitric oxide and thereby further trigger apoptosis and inflammatory responses in the bystander HUVECs through a p38-dependent pathway.

Topics: Atherosclerosis; Benzoates; Bystander Effect; Coculture Techniques; Human Umbilical Vein Endothelial Cells; Humans; Imidazoles; Inflammation; Isothiuronium; Macrophages; MAP Kinase Signaling System; Nitric Oxide; p38 Mitogen-Activated Protein Kinases; Pyridines; U937 Cells

Renal injury induced by brain death is characterized by ischemia and inflammation, and limiting it is a therapeutic goal that could improve outcomes in kidney transplantation. Brain death resulted in decreased circulating nitrite levels and increased infiltrating inflammatory cell infiltration into the kidney. Since nitrite stimulates nitric oxide signaling in ischemic tissues, we tested whether nitrite therapy was beneficial in a rat model of brain death followed by kidney transplantation. Nitrite, administered over 2 h of brain death, blunted the increased inflammation without affecting brain death-induced alterations in hemodynamics. Kidneys were transplanted after 2 h of brain death and renal function followed over 7 days. Allografts collected from nitrite-treated brain-dead rats showed significant improvement in function over the first 2 to 4 days after transplantation compared with untreated brain-dead animals. Gene microarray analysis after 2 h of brain death without or with nitrite therapy showed that the latter significantly altered the expression of about 400 genes. Ingenuity Pathway Analysis indicated that multiple signaling pathways were affected by nitrite, including those related to hypoxia, transcription, and genes related to humoral immune responses. Thus, nitrite therapy attenuates brain death-induced renal injury by regulating responses to ischemia and inflammation, ultimately leading to better post-transplant kidney function.

Topics: Allopurinol; Animals; Benzoates; Brain Death; Gene Expression; Hemodynamics; Imidazoles; Inflammation; Kidney; Kidney Transplantation; Lipid Peroxidation; Male; Nitrites; Rats; Rats, Inbred Lew; Reperfusion Injury; Signal Transduction; Sodium Nitrite

Six azaphilones, monascin (1), ankaflavin (2), rubropunctatin (3), monascorburin (4), rubropunctamine (5), and monascorburamine (6), two furanoisophthalides, xanthomonasin A (7) and xanthomonasin B (8), and two amino acids, (+)-monascumic acid (9) and (-)-monascumic acid (10), isolated from the extracts of Monascus pilosus-fermented rice (red-mold rice) were evaluated for their inhibitory effects on 12-O-tetradecanoylphorbol-13-acetate (TPA)-induced inflammation in mice, on the induction of Epstein-Barr virus early antigen (EBV-EA) by TPA in Raji cells, and on the activation of (+/-)-(E)-methyl-2[(E)-hydroxy-imino]-5-nitro-6-methoxy-3-hexemide (NOR 1), a nitric oxide (NO) donor. Among the compounds tested, seven compounds (1-6 and 10) on TPA-induced inflammation, and six compounds (1, 3-5, 9, and 10) on EBV-EA activation, exhibited potent inhibitory effects. All of the compounds tested showed moderate inhibitory effects on NOR 1 activation.

Topics: Animals; Antigens, Viral; Benzoates; Benzopyrans; Dicarboxylic Acids; Female; Fermentation; Imidazoles; Inflammation; Mice; Mice, Inbred ICR; Monascus; Oryza; Pigments, Biological; Xanthones

Aromatic amines, such as benzidine (BZ), initiate bladder cancer in humans. Inflammation/infection play an important role in this cancer. This study was designed to assess the influence of inflammatory oxidants, including reactive nitrogen oxygen species (RNOS), on BZ transformation and activation. RNOS were generated under various conditions and reacted with BZ, and the products were examined by HPLC. Conditions that generate nitrogen dioxide radical, NO(2)(-) + myeloperoxidase + H(2)O(2) and ONOO(-), produced primarily a single new product, which was identified by MS as azobenzidine (AZO-BZ). The myeloperoxidase-catalyzed reaction was inhibited by 1 mM cyanide and did not require NO(2)(-). Chloride (100 mM) reduced the myeloperoxidase reaction by 30% with taurine having little effect. In contrast, conditions that generate N(2)O(3), i.e., NO donor diethylamine (DEA) NONOate, produced two products, which were identified by MS as 4'-OH-4-aminobiphenyl (4'-OH-ABP) and 4-aminobiphenyl (ABP). 2-(4-Carboxyphenyl)-4,4,5,5-tetramethylimidazoline-1-oxyl-3-oxide, an oxidant of NO thought to produce NO(2)(*), had a biphasic effect on product formation. At a concentration equal to DEA NONOate, a 5-fold increase in BZ nitrosation was observed, while at higher concentrations nitrosation was greatly diminished and formation of AZO-BZ occurred. Glutathione prevented RNOS transformation of BZ. With MPO and ONOO(-), a new product was formed that cochromatographed with 3-(glutathione-S-yl)BZ. Glutathione also prevented nitrosation of BZ but did not form additional BZ products. HOCl-mediated activation of BZ, 4'-OH-ABP, and ABP to bind DNA was assessed. A higher level of binding was observed at pH 5.5 than pH 7.4. BZ elicited the most binding. More binding was observed at both pH values with 4'-OH-ABP than ABP. Thus, components of the inflammatory response are capable of BZ transformation and activation.

Topics: Animals; Benzidines; Benzoates; Biotransformation; Cattle; DNA; Glutathione; Hydrogen Peroxide; Hypochlorous Acid; Imidazoles; Inflammation; Mass Spectrometry; Oxidants; Oxidation-Reduction; Reactive Nitrogen Species; Reactive Oxygen Species

The key player for adaptation to reduced oxygen availability is the transcription factor hypoxia-inducible factor 1 (HIF-1), composed of the redox-sensitive HIF-1alpha and the constitutively expressed HIF-1beta subunits. Under normoxic conditions, HIF-1alpha is rapidly degraded, whereas hypoxia, CoCl(2), or desferroxamine promote protein stabilization, thus evoking its transcriptional activity. Because HIF-1 is regulated by reactive oxygen species, investigation of the impact of reactive nitrogen species was intended. By using different nitric oxide (NO) donors, dose- and time-dependent HIF-1alpha accumulation in close correlation with the release of NO from chemically distinct NO donors was established. Intriguingly, small NO concentrations induced a faster but transient HIF-1alpha accumulation than higher doses of the same NO donor. In contrast, NO attenuated up-regulation of HIF-1alpha evoked by CoCl(2) in a concentration- and time-dependent manner, whereas the desferroxamine-elicited HIF-1alpha signal remained unaltered. To demonstrate an autocrine or paracrine signaling function of NO, we overexpressed the inducible NO synthase and used a coculture system of activated macrophages and tubular cells. Expression of the NO synthase induced HIF-1alpha accumulation, which underscored the role of NO as an intracellular activator for HIF-1. In addition, macrophage-derived NO triggered HIF-1alpha up-regulation in LLC-PK(1) target cells, which points to intercellular signaling properties of NO in achieving HIF-1 accumulation. Our results show that NO does not only modulate the HIF-1 response under hypoxic conditions, but it also functions as a HIF-1 inducer. We conclude that accumulation of HIF-1 occurs during hypoxia but also under inflammatory conditions that are characterized by sustained NO formation.

Topics: Animals; Benzoates; Cell Hypoxia; Cell Line; Cobalt; Coculture Techniques; Deferoxamine; DNA-Binding Proteins; Dose-Response Relationship, Drug; Enzyme Inhibitors; Gene Expression Regulation; Glutathione; Guanylate Cyclase; Hydrazines; Hypoxia-Inducible Factor 1; Hypoxia-Inducible Factor 1, alpha Subunit; Imidazoles; Inflammation; Kidney Tubules, Proximal; Macrophage Activation; Macrophages; Mice; NG-Nitroarginine Methyl Ester; Nitric Oxide; Nitric Oxide Donors; Nitric Oxide Synthase; Nitric Oxide Synthase Type II; Nitrogen Oxides; Nitroso Compounds; Nuclear Proteins; Oxadiazoles; Oxazines; S-Nitrosoglutathione; Spermine; Swine; Transcription Factors

Recently, it has been reported that inflammatory processes are associated with the pathophysiology of Alzheimer's disease and that treatment of non-steroidal anti-inflammatory drugs reduce the risk for Alzheimer's disease. In the present study, we examined nitric oxide radical quenching activity of non-steroidal anti-inflammatory drugs and steroidal drugs using our established direct in vitro nitric oxide radical detecting system by electron spin resonance spectrometry. The non-steroidal anti-inflammatory drugs, aspirin, mefenamic acid, indomethacin and ketoprofen directly and dose-dependently scavenged generated nitric oxide radicals. In experiments of nitric oxide radical donor, NOC18-induced neuronal damage, these four non-steroidal drugs significantly prevented the NOC18-induced reduction of cell viability and apoptotic nuclear changes in neuronal cells without affecting the induction of inducible nitric oxide synthase-like immunoreactivity. However, ibuprofen, naproxen or steroidal drugs, which had less or no scavenging effects in vitro, showed almost no protective effects against NOC18-induced cell toxicity. These results suggest that the protective effects of the former four non-steroidal anti-inflammatory drugs against apoptosis might be mainly due to their direct nitric oxide radical scavenging activities in neuronal cells. These direct NO. quenching activities represent novel effects of non-steroidal anti-inflammatory drugs. Our findings identified novel pharmacological mechanisms of these drugs to exert not only their anti-inflammatory, analgesic, antipyretic activities but also neuroprotective activities against neurodegeneration.

Topics: Alzheimer Disease; Animals; Anti-Inflammatory Agents, Non-Steroidal; Aspirin; Benzoates; Cell Death; Cell Line; Cell Nucleus; Cell Survival; Dexamethasone; Electron Spin Resonance Spectroscopy; Free Radical Scavengers; Free Radicals; Humans; Hydrocortisone; Imidazoles; Indomethacin; Inflammation; Mefenamic Acid; Naproxen; Neuroblastoma; Neurons; Neuroprotective Agents; Nitric Oxide; Nitric Oxide Donors; Nitroso Compounds; Rats; Tumor Cells, Cultured