RESUMEN
BACKGROUND: Hypoxia-inducible factor-1 α (HIF-1 α ) and NF- κ B play important roles in the inflammatory response after hemorrhagic shock and resuscitation (H/R). Here, the role of myeloid HIF-1 α in liver hypoxia, injury, and inflammation after H/R with special regard to NF- κ B activation was studied. METHODS: Mice with a conditional HIF-1 α knockout (KO) in myeloid cell-line and wild-type (WT) controls were hemorrhaged for 90 min (30 ± 2 mm Hg) and resuscitated. Controls underwent only surgical procedures. RESULTS: After six hours, H/R enhanced the expression of HIF-1 α -induced genes vascular endothelial growth factor (VEGF) and adrenomedullin (ADM). In KO mice, this was not observed. H/R-induced liver injury in HIF-1 α KO was comparable to WT. Elevated plasma interleukin-6 (IL-6) levels after H/R were not reduced by HIF-1 α KO. Local hepatic hypoxia was not significantly reduced in HIF-1 α KO compared to controls after H/R. H/R-induced NF- κB phosphorylation in liver did not significantly differ between WT and KO. CONCLUSIONS: Here, deleting HIF-1 α in myeloid cells and thereby in Kupffer cells was not protective after H/R. This data indicates that other factors, such as NF- κB, due to its upregulated phosphorylation in WT and KO mice, contrary to HIF-1 α, are rather key modulators of inflammation after H/R in our model.
Asunto(s)
Hemorragia/metabolismo , Subunidad alfa del Factor 1 Inducible por Hipoxia/deficiencia , Inflamación/patología , Hepatopatías/inmunología , Animales , Femenino , Hemorragia/genética , Hipoxia/genética , Hipoxia/metabolismo , Subunidad alfa del Factor 1 Inducible por Hipoxia/genética , Inflamación/genética , Hepatopatías/etiología , Hepatopatías/genética , Ratones , Ratones NoqueadosRESUMEN
Gentamicin and cisplatin are clinically widely used pharmacological agents which may induce irreversible hearing loss as a side effect. Concerning the pathomechanisms of ototoxicity as well as preventive strategies there are similarities but also some differences. In this review we focus on the role of reactive oxygen species, the antioxidant system, cellular iron and calcium as well as nitric oxide and neurotrophins on gentamicin- and cisplatin-ototoxicity. Furthermore we deal with apoptotic and necrotic cell death as well as the role of mitochondria in these cell injury processes.
Asunto(s)
Aminoglicósidos/toxicidad , Antibacterianos/toxicidad , Antineoplásicos/toxicidad , Cisplatino/toxicidad , Sordera/inducido químicamente , Gentamicinas/toxicidad , Animales , Apoptosis/efectos de los fármacos , Relación Dosis-Respuesta a Droga , Cobayas , Células Ciliadas Auditivas/efectos de los fármacos , Humanos , Especies Reactivas de Oxígeno/metabolismo , Ganglio Espiral de la Cóclea/efectos de los fármacosRESUMEN
Aminoglycosides may induce irreversible hearing loss in both animals and humans. In order to study the nature and mechanisms underlying gentamicin-induced cell death in the inner ear, the cochlear neurosensory epithelia were dissected from guinea pigs and incubated with 0.5-10 mM gentamicin. Concentration-dependent loss of cell viability was detected by the inability of damaged cells to exclude propidium iodide. Outer hair cells were most sensitive towards gentamicin toxicity, followed by inner hair cells whereas Deiters and Hensen cells were not affected by the gentamicin concentrations used. The iron chelators 2,2'-dipyridyl and deferoxamine provided partial protection against gentamicin-induced hair cell death while the calcium chelator Quin-2 AM had no effect. Gentamicin (0.5-1 mM) induced condensation of chromatin typical for apoptosis. Using the fluorescent dye tetramethyl-rhodamine methyl ester and laser scanning microscopy we could visualize a loss of the mitochondrial membrane potential in damaged outer hair cells about 1 h before cell death occurred. Cyclosporin A, an inhibitor of the mitochondrial permeability pore, provided partial protection against gentamicin toxicity. This strongly suggests an involvement of the mitochondrial permeability transition in gentamicin-induced apoptosis.
Asunto(s)
Antibacterianos/toxicidad , Cóclea/efectos de los fármacos , Gentamicinas/toxicidad , Animales , Apoptosis/efectos de los fármacos , Quelantes/farmacología , Cóclea/metabolismo , Cóclea/patología , Ciclosporina/farmacología , Femenino , Cobayas , Células Ciliadas Auditivas Internas/efectos de los fármacos , Células Ciliadas Auditivas Internas/metabolismo , Células Ciliadas Auditivas Internas/patología , Células Ciliadas Auditivas Externas/efectos de los fármacos , Células Ciliadas Auditivas Externas/metabolismo , Células Ciliadas Auditivas Externas/patología , Humanos , Quelantes del Hierro/farmacología , Masculino , Mitocondrias/efectos de los fármacos , Mitocondrias/metabolismo , Permeabilidad , Especies Reactivas de Oxígeno/metabolismoRESUMEN
BACKGROUND: In extremely-low-birth-weight (ELBW) infants, formula feeding is required if human milk is not available. The tolerance of a new 'high' lactose (55 g/L), low protein, low phosphate, hydrolyzed protein formula (HLF) for early enteral feeding advancement of ELBW infants was compared with that of a low lactose (1 g/L) hydrolyzed protein formula (LLF). METHODS: In a randomized multicenter trial, 99 ELBW infants were fed according to a standardized protocol beginning at 48 hours of age with 12 ml/kg daily increments. Primary outcome was the cumulative milk feeding volume (CFV) from days 3 to 14. The authors hypothesized that feeding HLF as a supplement to human milk would increase the CFV at least by 20% in at least 60% of matched pairs compared with LLF. A secondary issue was to investigate whether human milk would increase the CFV compared with formula. RESULTS: The CFV was 720 mL/kg (range, 0-962 mL/kg) with HLF and 613 mL/kg (range, 3-1,283 mL/kg) with LLF feeding. There was no 20% difference. On day 14, the median feeding volume was 103 mL/kg. The CFV was 533 mL/kg (range, 0-962 mL/kg) in infants who received less than 10% of human milk and 832 mL/kg (range, 74-1,283 mL/kg) in infants who received more than 10%. Necrotizing enterocolitis (Bell stage > or =2) occurred only with LLF feeding (n = 5; P < 0.05). CONCLUSIONS: The study failed to find the hypothesized 20% advantage of the new HLF. The observed advantage of human milk supports the hypothesis that it should be the first diet in ELBW infants; however, this hypothesis still must be confirmed in a controlled, randomized trial.
Asunto(s)
Alimentos Infantiles , Recién Nacido de muy Bajo Peso/crecimiento & desarrollo , Lactosa/administración & dosificación , Leche Humana , Nutrición Enteral , Femenino , Humanos , Fenómenos Fisiológicos Nutricionales del Lactante , Recién Nacido , Recién Nacido de muy Bajo Peso/metabolismo , Unidades de Cuidado Intensivo Neonatal , Masculino , Distribución Aleatoria , Aumento de PesoRESUMEN
Since there are indications that iron influences cisplatin nephrotoxicity, we studied the role of iron in cisplatin ototoxicity in an in vitro model of the neurosensory epithelium of the guinea pig cochlea. Viability tests showed that Deiters and Hensen cells were not damaged and inner hair cells were only slightly damaged by cisplatin (50 microM). The outer hair cells were most sensitive to cisplatin toxicity. The iron chelator 2,2'-dipyridyl provided partial protection against cisplatin-induced cell death. In addition, we studied the influence of the iron chelators 2,2'-dipyridyl and deferoxamine on the chelatable iron pool in the various cells of the neurosensory epithelium using the fluorescent iron indicator Phen Green SK. Both chelators decreased the chelatable iron accessible to Phen Green SK, although the effect of deferoxamine was weaker because it entered the cells more slowly. The cellular concentration of the chelatable iron was measured using Phen Green SK and quantitative laser scanning microscopy. The concentration of chelatable iron in the inner ear cells ranged from 1.3 +/- 0.4 microM iron in inner hair cells to 3.7 +/- 1.7 microM iron in Hensen cells and did not correlate with the various cell types' susceptibility to cisplatin. Furthermore, cisplatin did not raise the intracellular chelatable iron concentration but enhanced the production of superoxide anions inside the neurosensory epithelium, especially inside the hair cells, as detected by the nitrotetrazolium blue reduction assay. Our conclusion is that cisplatin ototoxicity is partially mediated by an iron-dependent pathway and is associated with an enhanced formation of superoxide anions.
Asunto(s)
Antineoplásicos/toxicidad , Cisplatino/toxicidad , Sordera/inducido químicamente , Hierro/metabolismo , Superóxidos/metabolismo , 2,2'-Dipiridil/farmacología , Aminoquinolinas/farmacología , Animales , Calcio/metabolismo , Muerte Celular/efectos de los fármacos , Quelantes/farmacología , Cóclea/efectos de los fármacos , Cóclea/inervación , Deferoxamina/farmacología , Epitelio/efectos de los fármacos , Femenino , Colorantes Fluorescentes/farmacología , Cobayas , Células Ciliadas Auditivas Internas/efectos de los fármacos , Células Ciliadas Auditivas Externas/efectos de los fármacos , Quelantes del Hierro/farmacología , Cinética , Masculino , Microscopía Fluorescente , Nitroazul de Tetrazolio , Compuestos Orgánicos , Oxidación-ReducciónRESUMEN
Reactive oxygen species (ROS) have been postulated to be involved in drug ototoxicity and noise-induced hearing loss. Hydrogen peroxide (H(2)O(2))-induced cell damage in the inner ear was investigated using the neurosensory epithelium of a guinea pig cochlea. Hair cells and supporting cells of the epithelium incubated in Hanks' balanced salt solution were viable up to 6 h. After 2 h of treatment with 0.2 mM H(2)O(2) about 85% of the outer hair cells lost their viability. In contrast inner hair cells slowly began to die after 2 h of H(2)O(2) treatment. The Deiters cells and Hensen cells did not show any signs of damage in the presence of H(2)O(2). Nifedipine, a calcium channel blocker, Quin-2 AM, an intracellular calcium chelator, and 2,2'-dipyridyl, a membrane-permeable iron chelator, all provided partial protection against H(2)O(2)-induced outer hair cell death. The combination of both chelators showed an additional protective effect. The antioxidants N-acetylcysteine and glutathione-monoethyl ester completely protected against H(2)O(2) damage. These results suggest that calcium, iron, and thiol homeostasis play a crucial role in hair cell death caused by H(2)O(2).
Asunto(s)
Cóclea/efectos de los fármacos , Peróxido de Hidrógeno/farmacología , 2,2'-Dipiridil/farmacología , Aminoquinolinas/farmacología , Animales , Bloqueadores de los Canales de Calcio/farmacología , Supervivencia Celular/efectos de los fármacos , Quelantes/farmacología , Cóclea/citología , Cóclea/fisiología , Células Epiteliales/efectos de los fármacos , Células Epiteliales/fisiología , Cobayas , Células Ciliadas Auditivas Internas/efectos de los fármacos , Células Ciliadas Auditivas Internas/fisiología , Células Ciliadas Auditivas Externas/efectos de los fármacos , Células Ciliadas Auditivas Externas/fisiología , Técnicas In Vitro , Fármacos Neuroprotectores/farmacología , Nifedipino/farmacología , Factores de TiempoRESUMEN
Using a rat model, we report here the duration of heavy drinking necessary to produce immunosuppression, the recovery time after such alcohol-induced immunosuppression, and the variations in immune response associated with varied amounts of alcohol consumption. Immune status was evaluated by means of delayed cutaneous hypersensitivity (DCH)-like responses to phytohemagglutinin. The daily consumption of 5 g of ethanol/kg body weight/d resulted in a prompt reduction in DCH-like responses which was significant by Day 3 (p = 0.03) and maximal by Day 11 (45% of baseline). These data were consistent with a similar reduction of migration inhibitory factor activity in spleen cells from ethanol fed rats. Cessation of ethanol resulted in a return to baseline within 4 days. In a second experiment ethanol was administered daily in doses ranging from 0.5 to 6.0 g/kg. Early (Day 5) low dose ethanol (0.5-2 g/kg) stimulated immune response (153-188% of baseline) while high dose (6.0 g/kg) suppressed (79% of baseline). Continued treatments resulted in a loss of stimulation at low dose and increased suppression at higher doses. The relationship of these animal studies to human binge drinking and the possible risk for infection in the alcoholic remains to be established.
Asunto(s)
Etanol/farmacología , Inmunidad Celular/efectos de los fármacos , Animales , Relación Dosis-Respuesta a Droga , Exudados y Transudados/inmunología , Cobayas , Hipersensibilidad Tardía/inmunología , Factores Inhibidores de la Migración de Leucocitos/metabolismo , Masculino , Ratas , Ratas Endogámicas , Factores de TiempoRESUMEN
It is often necessary to have a small animal model which permits the sequential evaluation of functional immune status over a period of time. We report here the in vivo, intradermal response to phytohemagglutinin which produces an area of induration that is histologically similar to a typical delayed cutaneous hypersensitivity response, and that provides fast, quantitative, reproducible results similar to those observed with standard but more laborious and variable in vitro tests of immune function. For small animal studies this has the advantage of permitting longitudinal evaluations over time without sacrificing the animal. Using phytohemagglutinin-microprotein (0.2 mg/0.1 ml), injected intradermally, a delayed cutaneous hypersensitivity-like response is induced which is maximal at 24 hr. When immune function was altered either by treatment with a chemical immunosuppressant (ethanol) or by hormonal manipulations (hypophysectomy and rat growth hormone), the delayed cutaneous hypersensitivity-like response (area of induration) correlated closely with both macrophage migration inhibitory factor changes (r = 0.98; P less than 0.001) and mixed lymphocyte reaction changes (r = 0.99; P less than 0.05). These observations suggest that this technique correlates well with standard in vitro measures of immune response and may thus permit an in vivo estimation of immune reactivity.