RESUMEN
OBJECTIVE: Lorecivivint (LOR; SM04690), an investigational Wnt pathway modulator, previously demonstrated patient-reported and radiographic outcome improvements vs placebo in clinically relevant subjects with moderate to severe knee osteoarthritis (OA). This study's objective was to identify effective LOR doses. DESIGN: Subjects in this 24-week, Phase 2b, multicenter, randomized, double-blind, placebo (PBO)-controlled trial received an intra-articular injection of 2 mL LOR (0.03, 0.07, 0.15, or 0.23 mg), PBO, or dry-needle sham. The primary efficacy endpoints were changes in Pain NRS [0-10], WOMAC Pain [0-100], WOMAC Function [0-100], and radiographic mJSW outcomes, which were measured using baseline-adjusted analysis of covariance at Week 24. Multiple Comparison Procedure-Modeling (MCP-Mod) was performed for dose modeling. RESULTS: In total, 695/700 subjects were treated. Pain NRS showed significant improvements vs PBO after treatment with 0.07 mg and 0.23 mg LOR at Weeks 12 (-0.96, 95% CI [-1.54, -0.37], P = 0.001; -0.78 [-1.39, -0.17], P = 0.012) and 24 (-0.70 [-1.34, -0.06], P = 0.031; -0.82 [-1.51, -0.12], P = 0.022). Additionally, 0.07 mg LOR significantly improved WOMAC Pain and Function subscores vs PBO at Week 12 (P = 0.04, P = 0.021), and 0.23 mg LOR significantly improved both WOMAC subscores at Week 24 (P = 0.031, P = 0.017). No significant differences from PBO were observed for other doses. No radiographic progression was observed in any group at Week 24. MCP-Mod identified 0.07 mg LOR as the lowest effective dose. CONCLUSION: This 24-week Phase 2b trial demonstrated the efficacy of LOR on PROs in knee OA subjects. The optimal dose for future studies was identified as 0.07 mg LOR.
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Antiinflamatorios/uso terapéutico , Imidazoles/uso terapéutico , Indazoles/uso terapéutico , Osteoartritis de la Rodilla/tratamiento farmacológico , Piridinas/uso terapéutico , Método Doble Ciego , Femenino , Humanos , Inyecciones Intraarticulares , Masculino , Persona de Mediana Edad , Osteoartritis de la Rodilla/diagnóstico por imagen , Dimensión del Dolor , Medición de Resultados Informados por el Paciente , RadiografíaRESUMEN
OBJECTIVE: Hip osteoarthritis (OA) is difficult to treat. Steroid injections reduce pain with short duration. With widespread adoption of office-based, image-guided injections, hyaluronic acid is a potentially relevant therapy. In the largest clinical trial to-date, we compared safety/efficacy of a single, 6-mL image-guided injection of hylan G-F 20 to saline in painful hip OA. METHOD: 357 patients were enrolled in a multicenter, double-blind, randomized saline placebo- controlled trial. Subjects were ≥35 years of age, with painful (Western Ontario and McMaster Universities Osteoarthritis Index [WOMAC]-A1:5.0-8.0; numeric rating scale [NRS]: 0-10) mild-to-moderate hip OA (Kellgren-Lawrence grade II/III) and minimal contralateral hip pain (WOMAC-A1 < 4). Outcome measures included "pain on walking" (WOMAC-A1 and -A), Patient Global Self-Assessment (PTGA), WOMAC-A1 responder rate (+≥2 points on NRS), and adverse events (AEs) over 26 weeks. RESULTS: 357 patients (hylan G-F 20 single:182; saline:175) were enrolled. Both groups demonstrated significant pain improvement from baseline over 26 weeks (P < 0.0001); saline-induced pain reduction was a remarkable 35%. WOMAC-A and PTGA scores also significantly improved (P < 0.0001). No statistically significant difference was observed between groups in WOMAC-A1 scores (hylan G-F 20 single:-2.19 ± 0.16; saline:-2.26 ± 0.17) or WOMAC-A1 responders (41-52%). Treatment-related AE rates at target hip were similar (hylan G-F 20 single:23 patients [12.8%]; saline:12 [7.0%]). Posthoc analysis found, despite protocol requirements, many patients had psychological (31%) or potential neuropathic pain (27.5%) conditions. CONCLUSION: A single 6-mL hylan G-F 20 injection or saline for painful hip OA resulted in similar, statistically significant/clinically relevant pain and function improvements up to 6 months following injection; no differences between hylan G-F 20 and saline placebo were observed.
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Ácido Hialurónico/análogos & derivados , Osteoartritis de la Cadera/tratamiento farmacológico , Viscosuplementos/administración & dosificación , Acetaminofén/uso terapéutico , Anciano , Antiinflamatorios no Esteroideos/uso terapéutico , Método Doble Ciego , Quimioterapia Combinada , Femenino , Humanos , Ácido Hialurónico/administración & dosificación , Ácido Hialurónico/efectos adversos , Ácido Hialurónico/uso terapéutico , Inyecciones Intraarticulares , Masculino , Persona de Mediana Edad , Osteoartritis de la Cadera/complicaciones , Dolor/etiología , Dimensión del Dolor/métodos , Solución Salina , Índice de Severidad de la Enfermedad , Viscosuplementos/efectos adversos , Viscosuplementos/uso terapéutico , CaminataRESUMEN
OBJECTIVE: To assess the safety, pharmacokinetics, and exploratory efficacy of SM04690, a novel Wnt pathway inhibitor, as a potential disease modifying treatment for knee osteoarthritis (OA). DESIGN: Subjects with Kellgren-Lawrence grade 2-3 knee OA were randomized in successive dose-escalation cohorts to receive a knee intra-articular (IA) injection with 0.03, 0.07, or 0.23 mg SM04690, or placebo (PBO) (4:1 ratio). Safety, pharmacokinetics, efficacy (WOMAC Total/Function/Pain, Pain VAS, Physician Global Assessment [MDGA], and OMERACT-OARSI Response), OA-related biomarker (P1NP, ß-CTX, and cartilage oligomeric matrix protein [COMP]), and radiographic/imaging data were collected at baseline and during 24-week follow-up. RESULTS: 61 subjects (SM04690 n = 50; PBO n = 11) enrolled. Two dose limiting toxicities (DLTs), increased pain following injection and paroxysmal tachycardia (also the single serious AE), were reported in the 0.07 mg cohort. A total of 72 AEs were reported; Sixteen (occurring in eight subjects) were considered related to study medication. There were three discontinuations; one due to an AE (0.03 mg cohort). Bone marrow edema (BME) remained constant for most subjects. No doses were excluded from further study due to DLT criteria. Plasma levels of SM04690 were below the limit of detection at all time points. At Week 24, improvements from baseline were seen in all cohorts for the exploratory measures WOMAC Total, WOMAC Function, WOMAC Pain, MDGA, Pain VAS, and OMERACT-OARSI response. Joint space width (JSW) improvement was observed in the 0.07 mg cohort (P = 0.02 vs PBO). CONCLUSION: SM04690 appeared safe and well tolerated, with no evidence of systemic exposure. Exploratory efficacy analyses suggested positive trends for measurements of OA pain, function and disease-modifying osteoarthritis drug (DMOAD) properties. CLINICALTRIALS. GOV REGISTRATION: NCT02095548.
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Antirreumáticos/efectos adversos , Antirreumáticos/farmacocinética , Imidazoles/efectos adversos , Imidazoles/farmacocinética , Indazoles/efectos adversos , Indazoles/farmacocinética , Osteoartritis de la Rodilla/tratamiento farmacológico , Piridinas/efectos adversos , Piridinas/farmacocinética , Vía de Señalización Wnt/efectos de los fármacos , Anciano , Antirreumáticos/administración & dosificación , Antirreumáticos/uso terapéutico , Relación Dosis-Respuesta a Droga , Método Doble Ciego , Femenino , Humanos , Imidazoles/administración & dosificación , Imidazoles/uso terapéutico , Indazoles/administración & dosificación , Indazoles/uso terapéutico , Inyecciones Intraarticulares , Masculino , Persona de Mediana Edad , Osteoartritis de la Rodilla/diagnóstico por imagen , Dimensión del Dolor/métodos , Piridinas/administración & dosificación , Piridinas/uso terapéutico , Radiografía , Índice de Severidad de la EnfermedadRESUMEN
BACKGROUND: The role of the immune system in the surveillance of the body for cancer cells is well established. Human tumor cells do not survive in mice with intact immune systems, but they propagate in athymic nude mice. Presumably, the lack of a thymus gland and consequent loss of T lymphocytes results in a seriously compromised immune system without adequate cell-mediated immunity and tumor surveillance. In patients infected with the human immunodeficiency virus (HIV), a progressive loss of cell-mediated immunity is associated with the development of malignancies and opportunistic infections. This effect may be exacerbated in patients who chronically consume alcohol. METHODS: Normal and alcoholic BALB/c mice were treated with a monoclonal antibody to deplete CD4(+) lymphocytes before orthotopic implantation of human lung adenocarcinoma xenografts. Tumor volume and weight were measured and compared between groups. RESULTS: The authors' data show that a single treatment of anti-CD4 antibody causes almost complete depletion of CD4(+) lymphocytes and permits the formation of large intrapulmonary human nonsmall lung carcinoma xenografts in 100% of treated mice. All control animals injected with heat-denatured antibody failed to produce tumors. Chronic alcohol consumption by CD4-depleted mice resulted in larger tumors, compared with mice that did not receive ethanol in their diet (P = 0.05). CONCLUSIONS: Depletion of CD4(+) lymphocytes allows for the orthotopic growth of human lung adenocarcinoma xenografts in BALB/c mice. Furthermore, the consumption of alcohol reduces the ability of the impaired immune system to reject tumors.
Asunto(s)
Adenocarcinoma/inmunología , Alcoholismo/inmunología , Antígenos CD4/inmunología , Carcinoma de Pulmón de Células no Pequeñas/inmunología , Infecciones por VIH/inmunología , Neoplasias Pulmonares/inmunología , Linfocitos T/inmunología , Adenocarcinoma/patología , Animales , Anticuerpos Monoclonales , Recuento de Linfocito CD4 , Carcinoma de Pulmón de Células no Pequeñas/patología , Femenino , Infecciones por VIH/complicaciones , Humanos , Inmunidad Celular/inmunología , Terapia de Inmunosupresión , Neoplasias Pulmonares/patología , Ratones , Ratones Endogámicos BALB C , Trasplante Heterólogo , Células Tumorales Cultivadas/trasplanteRESUMEN
BACKGROUND: Thyroid hormones are endocrine modulators of several vital processes that are crucial to tumor growth and differentiation. Several anecdotal reports in the literature suggest that some histologic types of carcinoma may remain in a dormant state for prolonged periods of time in patients with hypothyroidism, with eventual progression of the disease once the decreased thyroid function is identified and corrected. METHODS: Oral propylthiouracil (PTU) was used to induce hypothyroidism in athymic nude mice that were subsequently inoculated with lung adenocarcinoma and prostate adenocarcinoma cells. Mice were also treated with a combination of PTU and thyroxine, which resulted in hyperthyroid levels of T(4). RESULTS: Subcutaneous lung and prostate xenografts grew significantly more slowly in hypothyroid mice treated with PTU than in euthyroid or hyperthyroid mice, regardless of treatment with PTU. Tumors grew well in groups of mice that were changed from a hypothyroid state to a euthyroid state by withdrawal of oral PTU. Administration of PTU 3 weeks after tumor inoculation also caused the tumor growth to slow significantly compared with tumors in mice that did not receive PTU. Mice that received PTU and thyroxine had tumors that grew as well as the tumors in euthyroid control animals. CONCLUSIONS: Our study indicates that human lung and prostate tumors do not grow well in hypothyroid nude mice, and that rendering these animals euthyroid has a significant impact on the growth rate of these tumors. Furthermore, in vitro and in vivo data indicated that this was not a result of an interaction of the tumor cells with PTU, but rather a result of the hypothyroid state.
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Antitiroideos/farmacología , Hipotiroidismo/fisiopatología , Neoplasias Experimentales/fisiopatología , Propiltiouracilo/farmacología , Animales , División Celular , Humanos , Hipotiroidismo/inducido químicamente , Masculino , Ratones , Ratones Desnudos , Trasplante de Neoplasias , Neoplasias Experimentales/sangre , Neoplasias Experimentales/patología , Tiroxina/sangre , Tiroxina/efectos de los fármacos , Tiroxina/farmacología , Factores de Tiempo , Trasplante Heterólogo , Células Tumorales CultivadasRESUMEN
PURPOSE: Prostate cancer is the most common malignancy of males in the United States. Although the overall survival rate for early stage prostate cancer is good, if cancer recurs following curative therapies there is no adequate salvage therapy. Systemic chemotherapy has never been associated with any meaningful improvement in overall survival or overall objective benefit. There is a need to develop novel therapies for prostate cancer. MATERIALS AND METHODS: Two prostatic cancer cell lines, DU-145 and PC-3, were grown as subcutaneous xenografts in athymic nude mice. The recombinant oncotoxin AR209, formerly OLX-209 [e23(Fv)PE38KDEL]), has the specificity of an anti-p185erbB-2 antibody contained within a single-chain antibody domain (e23Fv) coupled to a portion of the Pseudomonas exotoxin A (PE38KDEL). Using Western blot analysis, the cell lines were shown to express p185erbB-2. The mice received either 3 i.v. injections, one every 2 days, of the recombinant oncotoxin AR209 or PBS, or were implanted with osmotic pumps that delivered a constant s.c. amount of AR209 or PBS. RESULTS: The oncotoxin was effective in reducing the size of s.c. prostatic xenografts in athymic nude mice. The data demonstrated that small tumors (<200 mm.3) were effectively reduced in size. However, larger tumors (>500 mm.3) were not effectively diminished. CONCLUSIONS: This study provides preliminary evidence for the utility of a recombinant oncotoxin in the treatment of prostate carcinoma. Recombinant oncotoxins may be an effective clinical addition for the management of metastatic prostate lesions in patients treated with conventional therapy.
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Adenocarcinoma/tratamiento farmacológico , Inmunotoxinas/uso terapéutico , Neoplasias de la Próstata/tratamiento farmacológico , Adenocarcinoma/metabolismo , Animales , Anticuerpos , Exotoxinas , Humanos , Masculino , Ratones , Ratones Desnudos , Trasplante de Neoplasias , Neoplasias de la Próstata/metabolismo , Receptor ErbB-2/biosíntesis , Proteínas Recombinantes de Fusión/uso terapéutico , Anticuerpos de Cadena Única , Trasplante HeterólogoRESUMEN
Lung cancer remains a significant public health problem in the U.S.A. and will result in an estimated 160,400 deaths in 1997. This appalling number is due in large part to the lack of adequate treatment for tumours that are refractory to surgery with curable intent, or of an adequate salvage therapy for those patients who recur after surgical resection. Because non-small cell lung cancer is refractory to traditional chemotherapy, non-traditional therapies have been developed to treat patients with this disease. Recombinant oncotoxins have been designed to target cells that express certain proteins as part of their cellular membrane. One such oncotoxin, AR209 (formerly OLX-209 [e23(Fv)PE38KDEL]), has the specificity of an anti-ErbB-2 antibody contained within a single-chain antibody domain (e23v) coupled to a portion of the Pseudomonas exotoxin A (PE38KDEL). Previous studies demonstrate that this drug is capable of significantly reducing the size of orthotopic lung tumour xenografts. However, most of the treated mice developed tumours once therapy was removed. In this study, mice were treated aggressively using one of four drug treatment schedules. Mice were treated with either intravenous or subcutaneous injections of AR209 over a 2 week period. The data indicate that AR209 significantly reduced the size of tumours and upon microscopic analysis at necropsy, some mice were cured. However, despite the treatment schedule used, many mice contained residual tumour. Residual tumours expressed the ErbB-2 protein, indicating that more aggressive treatment with AR209 may have resulted in higher rates of cure.
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Anticuerpos/uso terapéutico , Carcinoma de Pulmón de Células no Pequeñas/terapia , Genes erbB-2/inmunología , Neoplasias Pulmonares/terapia , Proteínas Recombinantes/uso terapéutico , Animales , Carcinoma de Pulmón de Células no Pequeñas/metabolismo , Esquema de Medicación , Femenino , Humanos , Inmunohistoquímica , Neoplasias Pulmonares/metabolismo , Ratones , Trasplante de Neoplasias , Neoplasia ResidualRESUMEN
RATIONALE AND OBJECTIVES: The authors performed this study to determine whether intrathoracic inoculation of non-small-cell lung carcinoma with fluoroscopic guidance would provide for more accurate implantation. MATERIALS AND METHODS: A tumor cell inoculum (2 x 10(6) cells per 0.15 mL) was injected percutaneously under fluoroscopic guidance at the posterior midaxillary line in 22 athymic nude mice. The mice underwent imaging with a mammographic unit at 3, 5, and 8 weeks after implantation. The mice were sacrificed at 8 weeks, and autopsy was performed to determine tumor yield. RESULTS: The use of a percutaneous technique under fluoroscopic guidance greatly facilitated the accurate implantation of xenografts. Tumor growth was seen at radiography in 18 of the 22 (82%) mice at 8 weeks. Necropsy revealed a 100% tumor yield. Histologic examination confirmed adenocarcinoma of the lung. The average number of tumors found in the lung parenchyma was 1.05 +/- 0.35; the average number of tumors found in the mediastinum was 0.59 +/- 0.67. The average tumor weight was 389 mg +/- 64.3. The average tumor size was 300 mm3 +/- 66.23. CONCLUSION: With fluoroscopic guidance, percutaneous implantation of tumor cells in athymic nude mice is simple and effective.
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Adenocarcinoma/diagnóstico por imagen , Carcinoma de Pulmón de Células no Pequeñas/diagnóstico por imagen , Fluoroscopía/métodos , Neoplasias Pulmonares/diagnóstico por imagen , Adenocarcinoma/patología , Animales , Carcinoma de Pulmón de Células no Pequeñas/patología , Modelos Animales de Enfermedad , Femenino , Neoplasias Pulmonares/patología , Ratones , Ratones Desnudos , Trasplante de Neoplasias , Células Tumorales CultivadasRESUMEN
The recombinant oncotoxin AR209 [e23(Fv)PE38KDEL; formerly OLX-209] was developed to treat neoplasia that expresses the c-erbB-2 (HER-2/neu) protein product p185(erbB-2). The AR209 compound contains a single-chain antibody domain specific for p185(erbB-2), coupled with a portion of the Pseudomonas exotoxin. The drug has been shown to be effective in inhibiting cells that overexpress erbB-2 due to gene amplification and in cells that do not contain amplified erbB-2 but express slightly higher levels of the protein than normal cells do. To test the efficacy of AR209 on human lung tumors, athymic nude mice were inoculated intrathoracically with a cell line derived from a poorly differentiated lung adenocarcinoma. This cell line, termed 201T, expresses moderately elevated levels of p185(erbB-2) 7.6-fold over normal bronchial epithelium. Mice treated with i.v. injections of AR209 for 5 weeks after orthotopic tumor implantation had smaller tumors and in 20% of cases showed no evidence of disease. The data from this study indicate that AR209 may be an effective treatment for patients with non-small cell lung cancers that express p185(erbB-2).
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Antineoplásicos/uso terapéutico , Carcinoma de Pulmón de Células no Pequeñas/tratamiento farmacológico , Inmunotoxinas/uso terapéutico , Neoplasias Pulmonares/tratamiento farmacológico , Receptor ErbB-2/antagonistas & inhibidores , Animales , Anticuerpos , Carcinoma de Pulmón de Células no Pequeñas/diagnóstico por imagen , Carcinoma de Pulmón de Células no Pequeñas/patología , Exotoxinas , Humanos , Neoplasias Pulmonares/diagnóstico por imagen , Neoplasias Pulmonares/patología , Ratones , Ratones Desnudos , Trasplante de Neoplasias , Radiografía , Proteínas Recombinantes de Fusión/uso terapéutico , Proteínas Recombinantes/uso terapéutico , Anticuerpos de Cadena Única , Células Tumorales CultivadasRESUMEN
Reactive oxygen species have been implicated in the gastrointestinal pathogenesis of septic and endotoxic shock. The objective of this study was to investigate the role of inducible nitric oxide synthase during endotoxin-induced formation of oxidants by cells of the small intestine. After intravenous Escherichia coli lipopolysaccharide (LPS) (1 mg/kg) injection, nitric oxide production was measured as nitrosyl complex formation in the ileum using electron paramagnetic resonance spectroscopy. Oxidative stress biomarkers were determined as duodenal mucosal-reduced thiols, the ileal lipid peroxidation and luminal free radical production using spin trapping methodology. Demonstration of nitrosyl complex formation commenced at 3 h and diminished 24 h post-LPS. Mucosal thiol levels were decreased at 3, 6, 12, and 18 h post-LPS treatment. At these time point, the ileal lipid peroxidation also increased as did luminal formation of hydroxyl radical adduct. Nitric oxide synthase inhibitors reversed the elevation of hydroxyl radical formation and reversed the decrease in mucosal-reduced thiol levels in the LPS-treated rats. Our data indicate that nitric oxide or its oxidant product(s), such as peroxynitrite, contribute to oxidative injury in the small intestine of rats treated with endotoxin.
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Intestino Delgado/efectos de los fármacos , Lipopolisacáridos/farmacología , Óxido Nítrico Sintasa/biosíntesis , Óxido Nítrico/fisiología , Estrés Oxidativo/efectos de los fármacos , Análisis de Varianza , Animales , Óxidos N-Cíclicos , Inducción Enzimática , Radicales Libres , Intestino Delgado/citología , Intestino Delgado/metabolismo , Masculino , Óxido Nítrico/biosíntesis , Ratas , Ratas Sprague-Dawley , Marcadores de SpinRESUMEN
The rate of glucose utilization (Rg) of various tissues including lung and bronchoalveolar lavage (BAL) fluids cells was measured, using the 2-deoxyglucose technique in Sprague-Dawley rats 4 h after challenge with either 1 mg/kg intravenous or 0.3 mg/kg intratracheal lipopolysaccharide (LPS). After intravenous LPS, Rg increased in whole lung and nonrespiratory tissues, but was unaltered in BAL cells. After intratracheal LPS, the Rg of nonrespiratory tissues was unchanged, but the Rg of BAL cells increased from 3.7 +/- 0.3 to 71.5 +/- 16.0 nmol/min. This increase in the Rg of BAL cells was explained by a doubling of the macrophage specific Rg, by a 100-fold increase in polymorphonuclear leukocytes (PMN) number, and by a higher Rg in PMN than in macrophages. These results demonstrate that increased glucose utilization after intratracheal LPS is confined to the respiratory system and that intra-alveolar phagocytes participate in this increase.
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Glucosa/metabolismo , Lipopolisacáridos/administración & dosificación , Pulmón/metabolismo , Animales , Glucemia/efectos de los fármacos , Glucemia/metabolismo , Presión Sanguínea/efectos de los fármacos , Líquido del Lavado Bronquioalveolar/química , Líquido del Lavado Bronquioalveolar/citología , Desoxiglucosa/metabolismo , Glucólisis/efectos de los fármacos , Frecuencia Cardíaca/efectos de los fármacos , Inyecciones Intravenosas , Intubación Intratraqueal , Lactatos/sangre , Lipopolisacáridos/farmacología , Pulmón/efectos de los fármacos , Macrófagos Alveolares/citología , Macrófagos Alveolares/efectos de los fármacos , Masculino , Neutrófilos/citología , Neutrófilos/efectos de los fármacos , Especificidad de Órganos , Ratas , Ratas Sprague-DawleyRESUMEN
The objective of this study was to demonstrate nitric oxide (NO) production and determine its role in the rat small intestine following endotoxin treatment. By using electron paramagnetic resonance (EPR) spectroscopy, we were able to detect high concentrations of nitrosylated proteins in the small intestines of rats administered 1 mg/kg lipopolysaccharide (LPS) and sacrificed 6 h later. EPR spectra of non-heme and heme nitrosyl complexes were detected in the epithelium layer and intestinal wall. Only EPR spectra characteristic of nitrosyl hemoprotein complexes were detected in the luminal contents of these rats. LPS administration elevated the concentrations of intestinal lipid peroxidation biomarkers, thiobarbituric acid-reactive substances, and conjugated dienes. These changes were attenuated by NO synthase inhibitor treatment. We conclude that oxidants associated with NO formation were at least in part involved in the oxidation of tissue lipids. This process may be one of the mechanisms of intestinal injury induced by LPS.
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Intestino Delgado/efectos de los fármacos , Peroxidación de Lípido/efectos de los fármacos , Lipopolisacáridos/farmacología , Óxido Nítrico/biosíntesis , Sustancias Reactivas al Ácido Tiobarbitúrico/metabolismo , Animales , Biomarcadores/química , Hemoglobinas/metabolismo , Intestino Delgado/metabolismo , Masculino , Peroxidasa/metabolismo , Ratas , Ratas Sprague-DawleyRESUMEN
The aim of the present work was to test the effect of acute in vivo alcohol administration (180-190 mg/dl plasma for 3 h) on glucose utilization by tissues under basal conditions or after a hyperinsulinemic (100-130 microU/ml) euglycemic clamp in fasted rats. In vivo glucose use by individual tissues was assessed by the tracer 2-deoxy-D-glucose technique. Alcohol administration to saline-infused rats markedly inhibited glucose use by skeletal muscles, including the soleus, white and red quadriceps, and gastrocnemius, as well as by the heart. Ethanol infusion, however, had no effect on glucose use by the diaphragm, lung, liver, skin, ileum, brain, and adipose tissue. The insulin-stimulated glucose use was also inhibited by alcohol selectively in the muscles, with no effect on other tissues tested, except a moderate inhibition in the brain. Ethanol inhibited muscle glucose use by an average of approximately 50% under both basal and insulin-stimulated conditions. However, because insulin treatment more than doubled basal glucose use by these muscles, the 50% inhibition by ethanol treatment represents a greater inhibition of absolute glucose use under insulin-stimulated rather than under basal conditions. Our data demonstrate that acute alcohol intake attenuates basal and hormone-induced glucose utilization in a tissue-specific fashion. The inhibitory effect of alcohol on skeletal muscle glucose use could contribute to the previously observed decreased glucose recycling in humans after acute alcohol intake.
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Etanol/toxicidad , Glucosa/metabolismo , Insulina/farmacología , Proteínas Musculares , Músculo Esquelético/metabolismo , Animales , Transportador de Glucosa de Tipo 4 , Masculino , Proteínas de Transporte de Monosacáridos/análisis , Ratas , Ratas Sprague-DawleyAsunto(s)
Ácido Clodrónico/administración & dosificación , Ácido Clodrónico/farmacología , Endotoxinas/farmacología , Macrófagos del Hígado/citología , Macrófagos/citología , Superóxidos/metabolismo , Análisis de Varianza , Animales , Basófilos/citología , Basófilos/metabolismo , Recuento de Células Sanguíneas , Separación Celular , Sistema Inmunológico/fisiología , Inmunohistoquímica , Macrófagos del Hígado/efectos de los fármacos , Macrófagos del Hígado/metabolismo , Leucocitos/citología , Lipopolisacáridos/farmacología , Liposomas , Macrófagos/efectos de los fármacos , Masculino , Modelos Biológicos , Neutrófilos/citología , Neutrófilos/metabolismo , Ratas , Ratas Sprague-Dawley , Factor de Necrosis Tumoral alfa/análisisRESUMEN
We tested the hypothesis that Kupffer cells modulate sinusoidal endothelial cell function in the liver. Rats were treated with Kupffer cell-depleting agents (gadolinium chloride and liposome-encapsulated dichloromethylene diphosphonate) or with inhibitors of phospholipase A2 or leukotriene A4 synthase (dexamethasone and diethylcarbamazine, respectively). Hyaluronan uptake by the isolated, perfused liver was measured as an index of the functional state of the sinusoidal endothelial cell. Plasma hyaluronan concentration was also determined. Three hours after Escherichia coli lipopolysaccharide administration (100 micrograms/100 gm body wt, intravenously) plasma hyaluronan levels were significantly increased (280% to 320%), whereas hepatic hyaluronan uptake was markedly decreased (approximately 76%). Pretreatment with gadolinium chloride (0.5 mg/100 gm body wt, intravenously, 21 hr before saline solution or lipopolysaccharide administration), liposome-encapsulated dichloromethylene diphosphonate (40 mumol/100 gm body wt, intravenously, 44 hr before saline solution or lipopolysaccharide injection), dexamethasone (40 micrograms/100 gm body wt, intravenously, 1 hr before saline solution or lipopolysaccharide administration) or diethylcarbamazine (repeated doses, 10 mg/100 gm body wt, intravenously, 1 hr before saline solution or lipopolysaccharide injection) counteracted the lipopolysaccharide inhibitory effect on hepatic hyaluronan uptake. With the exception of gadolinium chloride, all other agents also prevented the lipopolysaccharide-induced increase in plasma hyaluronan concentration. Gadolinium chloride only attenuated the lipopolysaccharide effect on plasma hyaluronan level. Taken together with earlier results from our laboratory, these data indicate that: (a) Kupffer cell activation by lipopolysaccharide results in suppression of hyaluronan uptake by sinusoidal endothelial cells and (b) such modulation of endothelial cell function is likely mediated by products of the lipoxygenase pathway of arachidonate metabolism.
Asunto(s)
Comunicación Celular , Macrófagos del Hígado/fisiología , Hígado/fisiología , Animales , Ácido Araquidónico/metabolismo , Ácido Clodrónico/administración & dosificación , Ácido Clodrónico/farmacología , Dexametasona/farmacología , Dietilcarbamazina/farmacología , Portadores de Fármacos , Endotelio/citología , Endotelio/fisiología , Escherichia coli , Gadolinio/farmacología , Ácido Hialurónico/sangre , Ácido Hialurónico/metabolismo , Macrófagos del Hígado/efectos de los fármacos , Lipopolisacáridos/farmacología , Liposomas , Hígado/citología , Masculino , Técnicas de Cultivo de Órganos , Ratas , Ratas Sprague-DawleyRESUMEN
The present study was performed to determine the time-course for the development of peripheral and hepatic insulin resistance in rats as a result of an increasing tumor burden. Animals were inoculated with Yoshida ascites hepatoma, and studies were conducted during the early phase of tumor growth (day 4) at which time there was no change in food intake and at a later time point (day 8) when the tumor burden was increased and rats demonstrated anorexia. In vivo insulin action was accessed under euglycemic hyperinsulinemic conditions, in which insulin was infused at rates sufficient to produce arterial insulin levels that represent high physiological (3.5 ng/ml) or maximally stimulating values (180 ng/ml). On day 4, tumor-bearing (TB) rats were euglycemic, and whole body glucose turnover was elevated 32%. Insulin-mediated glucose uptake (IMGU) in TB rats was similar to control values at the low insulin infusion rate but reduced by 53% under maximally stimulating conditions. The insulin-induced suppression of glucose production was similar in TB and control animals at this time point. In contrast, on day 8, TB rats were hypoglycemic and glucose turnover was reduced 35%. The impairment in IMGU was more severe than seen earlier, with glucose uptake being reduced 39 and 61% at both levels of hyperinsulinemia. At this time point, the ability of insulin to inhibit glucose production was also impaired. These results indicate that the insulin resistance induced by the Yoshida hepatoma was manifested initially by a reduction in IMGU by peripheral tissues. As the tumor burden increased peripheral insulin resistance became more severe and an impairment in hepatic insulin action was observed.
Asunto(s)
Glucosa/metabolismo , Insulina/farmacología , Neoplasias Hepáticas Experimentales/metabolismo , Neoplasias Hepáticas/metabolismo , Hígado/metabolismo , Animales , Peso Corporal , Ingestión de Alimentos , Hemodinámica , Cinética , Neoplasias Hepáticas/sangre , Neoplasias Hepáticas/patología , Neoplasias Hepáticas Experimentales/sangre , Neoplasias Hepáticas Experimentales/patología , Masculino , Trasplante de Neoplasias , Ratas , Ratas Sprague-DawleyRESUMEN
The purpose of this investigation was to examine the effect of gram-negative bacterial lipopolysaccharide on hyaluronan concentration in blood plasma, hyaluronan removal from the blood and hyaluronan uptake by isolated, perfused rat liver. Intravenous administration of Escherichia coli lipopolysaccharide to rats markedly increased plasma hyaluronan concentration in a dose-dependent manner. One day after lipopolysaccharide challenge (0.1 or 1.0 mg per 100 gm body wt), plasma hyaluronan levels were 570.7 +/- 66.8 ng x ml-1 and 1,951.0 +/- 120.3 ng x ml-1, respectively, as compared with 94.2 +/- 12.2 ng x ml-1 in the time-matched control animals. Removal of intravenously injected hyaluronan (30 micrograms per 100 gm body wt) was suppressed 32% by lipopolysaccharide administration (100 micrograms per 100 gm body wt). At the same dose, lipopolysaccharide induced a severe inhibition (60% to 80%) of hyaluronan uptake by perfused livers isolated 3 or 24 hr after lipopolysaccharide administration. The inhibitory effect of lipopolysaccharide on hyaluronan uptake by the isolated, perfused liver was not abolished by pretreatment with either antibodies to tumor necrosis factor-alpha IgG or indomethacin, an inhibitor of the cyclooxygenase pathway. Continuous intravenous infusion of recombinant murine tumor necrosis factor-alpha for 18 to 20 hr did not affect plasma hyaluronan concentration. These data suggest that neither tumor necrosis factor-alpha, an early cytokine induced by lipopolysaccharide, nor prostaglandins are involved in the mechanism of lipopolysaccharide-induced inhibition of hyaluronan uptake by the perfused rat liver.(ABSTRACT TRUNCATED AT 250 WORDS)
Asunto(s)
Escherichia coli , Ácido Hialurónico/metabolismo , Lipopolisacáridos/farmacología , Hígado/metabolismo , Animales , Anticuerpos , Ácido Hialurónico/sangre , Inmunoglobulina G , Indometacina/farmacología , Cinética , Masculino , Prostaglandinas/fisiología , Ratas , Ratas Sprague-Dawley , Proteínas Recombinantes/farmacología , Factor de Necrosis Tumoral alfa/inmunología , Factor de Necrosis Tumoral alfa/farmacología , Factor de Necrosis Tumoral alfa/fisiologíaRESUMEN
Increased hepatic glucose production and glucose utilization involving multiple tissues occur in response to administration of bacterial lipopolysaccharide (LPS) and are metabolic hallmarks of hypermetabolic sepsis. As a proximal mediator in the host response to infection-like challenges, tumor necrosis factor (TNF) may enhance glucose metabolism by directly interacting with cells or by initiating a cascade of events leading to changes in glucose production and utilization. To determine if endogenous TNF is an important mediator in LPS- or sepsis-induced changes in glucose metabolism, rats were pretreated with a neutralizing goat anti-TNF IgG antibody prior to intravenous LPS or subcutaneous live Escherichia coli administration. Whereas high levels of plasma TNF were observed in rats not pretreated with anti-TNF, TNF was not detected 90 min after LPS in rats receiving the antibody. Pretreatment with anti-TNF attenuated the increase in plasma lactate and glucagon levels in LPS-challenged rats but failed to ameliorate the LPS-induced hyperglycemia and increase in glucose rate of appearance (Ra). The LPS-stimulated increase of in vivo glucose metabolic rate (Rg) of examined tissues, measured with [14C]-2-deoxyglucose, was not altered by anti-TNF. Likewise, rats treated with anti-TNF prior to induction of hypermetabolic infection exhibited usual increases in whole-body glucose Ra and metabolic clearance rate. Although neutralizing TNF failed to prevent the sepsis-induced augmentation of Rg in any tissue examined, it reduced the increase in the lung (P < 0.05) and tended to decrease it in other barrier tissues as well as in the spleen.(ABSTRACT TRUNCATED AT 250 WORDS)
Asunto(s)
Endotoxinas/farmacología , Infecciones por Escherichia coli/metabolismo , Glucosa/metabolismo , Factor de Necrosis Tumoral alfa/fisiología , Animales , Inmunoglobulina G/inmunología , Inmunoglobulina G/farmacología , Cinética , Masculino , Ratas , Ratas Sprague-Dawley , Factor de Necrosis Tumoral alfa/inmunologíaRESUMEN
Administration of tumor necrosis factor (TNF-alpha) increases whole body glucose kinetics and stimulates in vivo glucose uptake by several tissues. Because circulating catecholamines are also increased after TNF-alpha administration, the present study was conducted to examine the potential role of the adrenergic system in eliciting these changes. Rats given 150 micrograms TNF-alpha/kg by intravenous infusion over a 30-min period exhibited an increased rate of glucose appearance (glucose Ra). Combined alpha- and beta-adrenergic blockade (phentolamine and propranolol infusion) prevented the TNF-alpha-induced increase in glucose Ra without influencing plasma glucagon or corticosterone levels. TNF-alpha infusion also increased in vivo glucose utilization (Rg), measured with 2-deoxy-[14C]glucose, in spleen (86%), liver (80%), skin (47%), ileum (71%), lung (53%), and heart (112%). Adrenergic blockade prevented the tissue Rg increase in the spleen, liver, and skin; partially reduced it in the ileum; but did not abrogate it in the lung or heart. The effect of blockade was primarily due to inhibition of the TNF-alpha-induced increase in hepatic glucose output. Whereas the adrenergic system plays a major role on the effect of TNF-alpha on whole body glucose production, its importance in directly mediating TNF-alpha's effect on tissue glucose uptake is minimal.
Asunto(s)
Glucosa/metabolismo , Simpaticolíticos/farmacología , Factor de Necrosis Tumoral alfa/farmacología , Animales , Glucemia/análisis , Cinética , Masculino , Tasa de Depuración Metabólica , Concentración Osmolar , Ratas , Ratas Endogámicas , Distribución Tisular , Factor de Necrosis Tumoral alfa/antagonistas & inhibidores , Factor de Necrosis Tumoral alfa/metabolismoRESUMEN
Epinephrine produces smaller incremental increases in plasma glucose concentration and rate of glucose appearance (Ra) in septic rats compared with nonseptic animals. In the present study, we investigated the role of insulin in the diminished response of septic rats to epinephrine-induced increases in glucose turnover. Glucose kinetics were assessed by the infusion of [6-3H]-glucose in conscious catheterized rats made septic by subcutaneous injections of live Escherichia coli. Epinephrine was infused at 1 micrograms/min/kg for 2 hours in the presence and absence of somatostatin and mannoheptulose (SRIF + MH). In comparison to nonseptic control animals, epinephrine-induced increases in plasma glucose concentration and glucose Ra were blunted by more than 50% in the septic rats. Infusion of SRIF + MH with epinephrine restored the blunted response to normal. During the infusion of epinephrine alone, the plasma insulin concentration in the septic rats was 2.8-fold higher than the nonseptic controls. SRIF + MH lowered the plasma insulin concentrations in both the nonseptic and septic rats to less than 10 microU/mL. SRIF + MH reversed the sepsis-induced hyperglucagonemia, but did not prevent a slight increase in glucagon levels during the epinephrine infusion in the nonseptic rats. In a second study, septic rats infused with SRIF + MH and replacement insulin showed a smaller increase in glucose concentration and glucose production in response to epinephrine than did septic animals administered SRIF + MH and no insulin. These results indicate that insulin plays an important role in the diminished response of septic rats to epinephrine.