5-5--6-6--tetrachloro-1-1--3-3--tetraethylbenzimidazolocarbocyanine and Inflammation

Topics: Animals; Benzimidazoles; Carbocyanines; Fluorescent Dyes; Gene Expression Regulation; Inflammation; Interleukin-6; Lipopolysaccharides; Membrane Potential, Mitochondrial; Mice; Microscopy, Fluorescence; Mitochondria; Models, Biological; NF-kappa B; Nitrates; Nitric Oxide; Nitric Oxide Synthase Type II; Organophosphorus Compounds; Phenanthridines; RAW 264.7 Cells; Reactive Oxygen Species; Tumor Necrosis Factor-alpha; Xanthine Dehydrogenase

Intracerebral hemorrhage (ICH) is a cerebrovascular disease with high mortality and morbidity, and the effective treatment is still lacking. We designed this study to investigate the therapeutic effects and mechanisms of melatonin on the secondary brain injury (SBI) after ICH. An in vivo ICH model was induced via autologous whole blood injection into the right basal ganglia in Sprague-Dawley (SD) rats. Primary rat cortical neurons were treated with oxygen hemoglobin (OxyHb) as an in vitro ICH model. The results of the in vivo study showed that melatonin alleviated severe brain edema and behavior disorders induced by ICH. Indicators of blood-brain barrier (BBB) integrity, DNA damage, inflammation, oxidative stress, apoptosis, and mitochondria damage showed a significant increase after ICH, while melatonin reduced their levels. Meanwhile, melatonin promoted further increasing of expression levels of antioxidant indicators induced by ICH. Microscopically, TUNEL and Nissl staining showed that melatonin reduced the numbers of ICH-induced apoptotic cells. Inflammation and DNA damage indicators exhibited an identical pattern compared to those above. Additionally, the in vitro study demonstrated that melatonin reduced the apoptotic neurons induced by OxyHb and protected the mitochondrial membrane potential. Collectively, our investigation showed that melatonin ameliorated ICH-induced SBI by impacting apoptosis, inflammation, oxidative stress, DNA damage, brain edema, and BBB damage and reducing mitochondrial membrane permeability transition pore opening, and melatonin may be a potential therapeutic agent of ICH.

Topics: Animals; Annexin A5; Antioxidants; Apoptosis; Benzimidazoles; Brain Edema; Brain Injuries; Carbocyanines; Cerebral Cortex; Cerebral Hemorrhage; Disease Models, Animal; DNA Damage; In Situ Nick-End Labeling; Inflammation; Male; Melatonin; Mitochondrial Diseases; Neurons; Oxidative Stress; Rats; Rats, Sprague-Dawley; Rhodamines; Time Factors

Globoid cell leukodystrophy or Krabbe disease (KD), is a hereditary disorder caused by galactosylceramidase deficiency. Progressive accumulation of psychosine is considered to be the critical pathogenetic mechanism of cell death in the Krabbe brain. Psychosine mechanism of action has not been fully elucidated. It seems to induce apoptosis in oligodendrocytes through a mitochondrial pathway and to up-regulate inflammatory cytokines production resulting in oligodendrocyte loss. Our aim was to evaluate the role of psychosine in apoptotic cell death and inflammatory response in a group of patients affected by KD using peripheral blood lymphocytes (PBLs) and peripheral blood mononuclear cells (PBMCs) as a cellular model. PBLs from KP and healthy controls were exposed to 20 microM psychosine and analysed by flow cytometry, agarose gel electrophoresis and fluorescence microscopy. Our results showed that psychosine induces apoptosis in PBLs through a mitochondrial pathway, but the apoptotic response was quite low especially KP. The role of psychosine in the up-regulation of cytokines (TNFalpha, IL8 and MCP1) has been evaluated by ELISA in PBMCs from KP and controls after stimulation with LPS and phytohemagglutinin. Both in basal condition and after LPS stimulation, cells from KP showed a significant increase in TNF-alpha production, reduced MCP1 levels and no modification in IL8. These results indicate that lymphomonocytes from KP had a basal proinflammatory pattern that was amplified by psychosine. In conclusion, the reduced apoptotic response and the atypical cytokine production observed in our experiments, suggest an involvement of inflammatory pattern in immune peripheral cells of KP.

Topics: Adult; Annexin A5; Apoptosis; Benzimidazoles; Carbocyanines; Case-Control Studies; Caspases; Cells, Cultured; Chemokine CCL2; Cytokines; Electrophoresis, Agar Gel; Enzyme Activation; Enzyme-Linked Immunosorbent Assay; Female; Flow Cytometry; Fluorescent Dyes; Humans; Inflammation; Interleukin-8; Leukocytes, Mononuclear; Leukodystrophy, Globoid Cell; Lipopolysaccharides; Male; Membrane Potential, Mitochondrial; Microscopy, Fluorescence; Mitochondria; Phytohemagglutinins; Psychosine; Time Factors; Tumor Necrosis Factor-alpha