RESUMO
BACKGROUND: Monoamine reuptake inhibitors exhibit unique clinical profiles that reflect distinct engagement of the central nervous system (CNS) transporters. METHODS: We used a translational strategy, including rodent pharmacokinetic/pharmacodynamic modeling and positron emission tomography (PET) imaging in humans, to establish the transporter profile of TD-9855, a novel norepinephrine and serotonin reuptake inhibitor. RESULTS: TD-9855 was a potent inhibitor of norepinephrine (NE) and serotonin 5-HT uptake in vitro with an inhibitory selectivity of 4- to 10-fold for NE at human and rat transporters. TD-9855 engaged norepinephrine transporters (NET) and serotonin transporters (SERT) in rat spinal cord, with a plasma EC50 of 11.7 ng/mL and 50.8 ng/mL, respectively, consistent with modest selectivity for NET in vivo. Accounting for species differences in protein binding, the projected human NET and SERT plasma EC50 values were 5.5 ng/mL and 23.9 ng/mL, respectively. A single-dose, open-label PET study (4-20mg TD-9855, oral) was conducted in eight healthy males using the radiotracers [(11)C]-3-amino-4- [2-[(di(methyl)amino)methyl]phenyl]sulfanylbenzonitrile for SERT and [(11)C]-(S,S)-methylreboxetine for NET. The long pharmacokinetic half-life (30-40 h) of TD-9855 allowed for sequential assessment of SERT and NET occupancy in the same subject. The plasma EC50 for NET was estimated to be 1.21 ng/mL, and at doses of greater than 4 mg the projected steady-state NET occupancy is high (>75%). After a single oral dose of 20mg, SERT occupancy was 25 (±8)% at a plasma level of 6.35 ng/mL. CONCLUSIONS: These data establish the CNS penetration and transporter profile of TD-9855 and inform the selection of potential doses for future clinical evaluation.
Assuntos
Inibidores da Captação de Neurotransmissores/farmacologia , Inibidores da Captação de Neurotransmissores/farmacocinética , Éteres Fenílicos/farmacologia , Éteres Fenílicos/farmacocinética , Piperidinas/farmacologia , Piperidinas/farmacocinética , Adulto , Compostos de Anilina , Animais , Análise Química do Sangue , Encéfalo/diagnóstico por imagem , Encéfalo/efeitos dos fármacos , Encéfalo/metabolismo , Meia-Vida , Humanos , Imageamento por Ressonância Magnética , Masculino , Modelos Biológicos , Morfolinas , Proteínas da Membrana Plasmática de Transporte de Norepinefrina/metabolismo , Tomografia por Emissão de Pósitrons , Compostos Radiofarmacêuticos , Ratos Sprague-Dawley , Reboxetina , Proteínas da Membrana Plasmática de Transporte de Serotonina/metabolismo , Medula Espinal/efeitos dos fármacos , Medula Espinal/metabolismo , SulfetosRESUMO
Tiotropium is currently the only once-daily, long-acting muscarinic antagonist (LAMA) approved in the United States and other countries for the treatment of chronic obstructive pulmonary disease (COPD). Glycopyrronium has shown promise as a LAMA and was recently approved for once-daily maintenance treatment of COPD in the European Union. Here, we describe the in vivo preclinical efficacy and lung selectivity of a novel inhaled muscarinic antagonist, TD-4208 (biphenyl-2-ylcarbamic acid 1-(2-{[4-(4-carbamoylpiperidin-1-ylmethyl)benzoyl]methylamino}ethyl)piperidin-4-yl ester) and compare its profile to tiotropium and glycopyrronium. In anesthetized dogs, TD-4208, along with tiotropium and glycopyrronium, produced sustained inhibition of acetylcholine-induced bronchoconstriction for up to 24 hours. In anesthetized rats, inhaled TD-4208 exhibited dose-dependent 24-hour bronchoprotection against methacholine-induced bronchoconstriction. The estimated 24-hour potency (expressed as concentration of dosing solution) was 45.0 µg/ml. The bronchoprotective potencies of TD-4208 and tiotropium were maintained after 7 days of once-daily dosing, whereas glycopyrronium showed a 6-fold loss in potency after repeat dosing. To assess systemic functional activity using a clinically relevant readout, the antisialagogue effect of compounds was also evaluated. The calculated lung selectivity index (i.e., ratio of antisialagogue and bronchoprotective potency) of TD-4208 was superior to glycopyrronium after both single and repeat dosing regimens and was superior to tiotropium after repeat dosing. In conclusion, the in vivo preclinical profile suggests that TD-4208 has the potential to be a long-acting bronchodilator for once-daily treatment of respiratory diseases. Its greater functional selectivity for the lung in preclinical models may translate to an improved tolerability profile compared with marketed muscarinic receptor antagonists.
Assuntos
Benzamidas/farmacologia , Broncodilatadores/farmacologia , Carbamatos/farmacologia , Pulmão/efeitos dos fármacos , Antagonistas Muscarínicos/farmacologia , Acetilcolina/farmacologia , Administração por Inalação , Animais , Broncoconstrição/efeitos dos fármacos , Broncodilatadores/administração & dosagem , Cães , Relação Dose-Resposta a Droga , Glicopirrolato/administração & dosagem , Glicopirrolato/farmacologia , Pulmão/fisiologia , Masculino , Cloreto de Metacolina , Antagonistas Muscarínicos/administração & dosagem , Pilocarpina , Ratos , Ratos Sprague-Dawley , Receptor Muscarínico M1/antagonistas & inibidores , Receptor Muscarínico M3/antagonistas & inibidores , Salivação/efeitos dos fármacos , Derivados da Escopolamina/administração & dosagem , Derivados da Escopolamina/farmacologia , Brometo de TiotrópioRESUMO
Orofacial pain is a common complaint with multiple diagnoses. There is controversy about the effectiveness of antidepressants for the management of orofacial pain disorders. In order to be able to make a best evidence choice between available antidepressants for the treatment of orofacial pain, a systematic review was conducted of existing randomized controlled trials of antidepressants. Trials were identified from the Pubmed database up to March 2012, from references in retrieved reports and from references in review articles. Six articles were found and included in this review. Four studies were randomized placebo-controlled trials and two studies were randomized active-controlled trials. Two independent investigators reviewed these articles using a 15-item checklist. All six trials were of high quality according to the 15-item criteria. Nevertheless there was limited evidence to support the effectiveness of antidepressants in orofacial pain disorders, because of the heterogeneity of treatment modalities and the low number of randomized controlled trials per diagnose. More randomized controlled trials are needed to come to a firm conclusion for the use of antidepressants for orofacial pain disorders.
Assuntos
Antidepressivos/uso terapêutico , Dor Facial/tratamento farmacológico , Humanos , Placebos , Ensaios Clínicos Controlados Aleatórios como AssuntoRESUMO
This study describes the comparison of multiple and single pain ratings in patients after surgical removal of the third molar. Correlation and agreement analysis were performed between the average pain intensity measured three times a day over a period of 7 days and one single pain rating (designated the 'recalled average' pain, as assessed by the patient) after surgical removal of the lower third molar. The study population consisted of patients participating in three randomized trials. The results show that both measurement correlated good to excellent, but there was a large variation in agreement between the ratings. In conclusion, in patients with surgical third molar removal a single pain rating is not an accurate predictor of the average pain measured by a multiple pain-rating test. A single pain rating cannot replace a multiple pain assessment.
Assuntos
Dente Serotino/cirurgia , Medição da Dor/métodos , Adulto , Feminino , Humanos , Masculino , Mandíbula/cirurgia , Medição da Dor/estatística & dados numéricos , Dor Pós-Operatória/classificação , Ensaios Clínicos Controlados Aleatórios como Assunto , Autorrelato , Extração Dentária , Adulto JovemRESUMO
Inadequate antiprotease activity in the lungs due to alpha-1-antitrypsin (A1AT) deficiency is a factor of early-onset emphysema. We propose a new approach to gene therapy that involves the intratracheal delivery of macrophages expressing human A1AT (hA1AT). Recombinant adeno-associated virus (rAAV) plasmids encoding the hA1AT gene were packaged into virions using 293 cells, and transgenic progeny virus was purified from the cells. The murine macrophage cell line J774A.1 was infected in vitro with the recombinant hA1AT rAAV virus. The hA1AT-producing macrophages were delivered intratracheally into mechanically ventilated C57BL/6J mice, a strain with low endogenous levels of A1AT. Transcription of hA1AT mRNA was detected in the transfected cells by RT-PCR, and protein expression was verified by immunohistochemistry. Levels of hA1AT in the cell culture medium and in the bronchoalveolar lavage (BAL) were assayed by ELISA. The concentration of hA1AT in J774A.1 cell-conditioned medium increased from undetectable levels prior to transfection, to 60 mg/l at 24 h post-transfection. At 1, 3 and 7 days after intratracheal delivery of transfected macrophages, hA1AT protein in BAL from C57BL/6J mice increased from undetectable levels to 2.5+/-0.9, 2.6+/-1.1 and 2.2+/-0.8 mg/l, respectively. These results suggest that airway delivery of macrophages overexpressing hA1AT may be an effective approach to enhance alveolar protection in A1AT deficiency.
Assuntos
Dependovirus/genética , Pulmão/metabolismo , Macrófagos Alveolares/virologia , Transfecção/métodos , Deficiência de alfa 1-Antitripsina/terapia , alfa 1-Antitripsina/metabolismo , Administração por Inalação , Animais , Líquido da Lavagem Broncoalveolar/citologia , Ensaio de Imunoadsorção Enzimática , Terapia Genética/métodos , Vetores Genéticos/administração & dosagem , Camundongos , Camundongos Endogâmicos C57BL , Enfisema Pulmonar/terapia , alfa 1-Antitripsina/administração & dosagem , Deficiência de alfa 1-Antitripsina/metabolismoRESUMO
The amygdala is a temporal lobe region that is implicated in emotional information processing. The amygdala also is associated with the processing and modulation of pain sensation. Recently, we demonstrated that in nonhuman primates, the amygdala is necessary for the full expression of cannabinoid-induced antinociception [J Neurosci 21 (2001) 8238]. The antinociceptive effect of the cannabinoid receptor agonist (R)-(+)-[2,3-dihydro-5-methyl-3-(4-morpholinylmethyl)pyrrolo(1,2,3-de)-1,4-benzoxazin-6-yl]-1-naphthalenylmethanone (WIN55,212-2) was significantly reduced in rhesus monkeys with large bilateral lesions of the amygdaloid complex. In the present study, we investigated the contribution of the amygdala to cannabinoid-induced antinociception in the rat. Using bilateral local microinjections of the GABA(A) receptor agonist muscimol, we inactivated neurons originating from the central nucleus of the amygdala (CeA) or basolateral nucleus of the amygdala (BLA). In rats injected with intra-CeA saline, the cannabinoid receptor agonist WIN55,212-2 produced dose-dependent antinociception on the noxious heat-evoked tail flick assay. In rats treated with intra-CeA muscimol, however, the antinociceptive effect of WIN55,212-2 was significantly reduced. Rats treated with intra-BLA muscimol showed no deficit in WIN55,212-2-induced antinociception. The effect of CeA inactivation on WIN55,212-2-induced suppression of prolonged pain in the formalin test also was tested. In rats treated with intra-CeA saline, WIN55,212-2 reduced the incidence of formalin-induced nociceptive behaviors and also reduced formalin-evoked c-fos expression in both superficial and deep laminae of the spinal cord dorsal horn. In rats treated with intra-CeA muscimol, however, these effects of WIN55,212-2 were significantly reduced. The results constitute the first causal data demonstrating the necessity of descending pain-modulatory circuitry (of which the CeA is a component) for the full expression of cannabinoid-induced antinociception in the rat. Furthermore, the results complement previous findings suggesting an overlap in neural circuitry activated by opioids and cannabinoids.
Assuntos
Tonsila do Cerebelo/fisiologia , Analgésicos/farmacologia , Canabinoides/farmacologia , Tonsila do Cerebelo/anatomia & histologia , Tonsila do Cerebelo/efeitos dos fármacos , Analgésicos/uso terapêutico , Animais , Benzoxazinas , Canabinoides/uso terapêutico , Contagem de Células , Relação Dose-Resposta a Droga , Formaldeído , Agonistas GABAérgicos/farmacologia , Imuno-Histoquímica , Masculino , Microinjeções , Morfolinas/administração & dosagem , Muscimol/farmacologia , Naftalenos/administração & dosagem , Proteínas Oncogênicas v-fos/metabolismo , Dor/induzido quimicamente , Dor/tratamento farmacológico , Dor/metabolismo , Medição da Dor/efeitos dos fármacos , Ratos , Ratos Sprague-Dawley , Tempo de Reação , Medula Espinal/anatomia & histologia , Medula Espinal/metabolismoRESUMO
Targeting lung tissue is nonselective due in part to the lack of specific cell-surface receptors identified on target lung cells. We used in vivo phage display to identify a panel of peptides that can bind selectively to lung epithelial cells with less binding to nonepithelial cells. By direct intratracheal instillation of phage libraries into the lung, we isolated and identified 143 individual phage clones. Three phage clones revealed enhanced binding to the lung in vitro and in vivo. These three identified peptides were synthesized and demonstrated selective binding to epithelial cells in lung tissue versus the control peptide. Further, the peptides specifically bound to freshly isolated type II alveolar epithelial cells compared with Hep2 cells. The results suggest that the airway phage display approach could be exploited for analyzing the molecular diversity in the lower respiratory tract.
Assuntos
Marcação de Genes/métodos , Terapia Genética/métodos , Biblioteca de Peptídeos , Alvéolos Pulmonares/metabolismo , Sítios de Ligação , Células Cultivadas , Clonagem Molecular , Endocitose , Células Epiteliais/metabolismo , Células Epiteliais/ultraestrutura , Humanos , Microscopia Confocal , Microscopia Eletrônica , Microscopia Imunoeletrônica , Alvéolos Pulmonares/ultraestruturaRESUMO
Bleomycin is a well-established anti-tumor drug. Its major untoward effect, pulmonary toxicity, has limited its usage. In this study, we used a DNA repair protein, yeast apurinic/apyrimidinic endonuclease (APN1) to reduce the toxicity of bleomycin on lung cells. A549 cells, an alveolar epithelial cell line, were transduced by MIEG3 retroviral vector encoding both enhanced green fluorescent protein (EGFP) and APN1. Transduced cells were sorted by fluorescent-activated cell sorter (FACS) analysis and were cloned. The APN1 expression of transduced A549 cell population and four selected clones expressing different levels of EGFP was confirmed by Northern, Western, and apurinic/apyrimidinic (AP) endonuclease activity analyses. The expression of APN1 was positively correlated with the expression of EGFP. The protective effect of APN1 against bleomycin was determined by single cell gel electrophoresis/Comet assay and by clonogenic survival assay following bleomycin treatment. The A549 population expressing APN1 showed a significant reduction of DNA damage in the presence of 20, 50, and 100 microg/ml bleomycin; similarly, the APN1-expressing A549 population also demonstrated increased survival in the presence of bleomycin compared with the vector-transduced A549 population. In selected clones, three of four APN1-expressing clones resulted in significantly improved cell survival. The current study suggests that the yeast DNA repair protein, APN1, can reduce bleomycin toxicity to target lung cells.
Assuntos
Bleomicina/toxicidade , Dano ao DNA , Reparo do DNA/fisiologia , Endodesoxirribonucleases/fisiologia , Mutagênicos/toxicidade , Alvéolos Pulmonares/efeitos dos fármacos , Proteínas de Saccharomyces cerevisiae/fisiologia , Bleomicina/farmacologia , Sistema Livre de Células , Células Cultivadas/efeitos dos fármacos , Células Cultivadas/enzimologia , Células Clonais/efeitos dos fármacos , Células Clonais/metabolismo , Ensaio de Unidades Formadoras de Colônias , Ensaio Cometa , DNA/efeitos dos fármacos , DNA/metabolismo , Enzimas Reparadoras do DNA , Relação Dose-Resposta a Droga , Endodesoxirribonucleases/biossíntese , Endodesoxirribonucleases/genética , Citometria de Fluxo , Genes Reporter , Proteínas de Fluorescência Verde , Humanos , Ferro/metabolismo , Proteínas Luminescentes/análise , Proteínas Luminescentes/genética , Mutagênicos/farmacologia , Oxirredução , Alvéolos Pulmonares/citologia , Alvéolos Pulmonares/enzimologia , Proteínas Recombinantes de Fusão , Proteínas de Saccharomyces cerevisiae/biossíntese , Proteínas de Saccharomyces cerevisiae/genética , TransfecçãoRESUMO
Reversal of immunodeficiency in the lung by gene therapy is limited in part by the difficulty of transfecting lung cells in vivo. Many options exist for successfully transfecting cells in vitro, but they are not easily adapted to the in vivo condition. To overcome this limitation, we transduced macrophages in vitro with the murine IFN-gamma (mIFN-gamma) gene and intratracheally delivered the macrophages to express mIFN-gamma in vivo. A recombinant retroviral vector pSF91 system was modified to encode mIFN-gamma and enhanced green fluorescent protein (EGFP). A murine macrophage cell line J774A.1 transduced with the retroviral supernatant increased secretion from undetectable levels to 131.6 +/- 4.2 microg/ml mIFN-gamma at 24 h in vitro. The mIFN-gamma-producing macrophages were intratracheally instilled into mechanically ventilated scid mice. mIFN-gamma levels in the bronchoalveolar lavage increased from undetectable levels at baseline to 158.8 +/- 5.1 pg/ml at 48 h (P < 0.001). Analysis of the lavaged cells for EGFP expression revealed that EGFP expression was directly proportional to the number of transduced macrophages instilled into the lung. Immune function was partially restored in the alveolar spaces of scid mice with evidence of enhanced MHC class II antigen expression and increased phagocytosis (P < 0.05). Tumor necrosis factor alpha was increased from undetectable at baseline to 103.5 +/- 11.4 pg/ml. In contrast, i.p. administration of the engineered macrophages did not enhance IFN-gamma levels in the lung. Our study suggests airway delivery of genetically engineered macrophages expressing mIFN-gamma gene can partially restore significant immune activity in the lungs of immunodeficient mice.
Assuntos
Interferon gama/genética , Macrófagos Alveolares/imunologia , Animais , Linhagem Celular , Feminino , Expressão Gênica , Engenharia Genética , Terapia Genética , Antígenos de Histocompatibilidade Classe II/metabolismo , Técnicas In Vitro , Injeções Intraperitoneais , Interferon gama/biossíntese , Pulmão/imunologia , Pulmão/patologia , Camundongos , Camundongos Endogâmicos C57BL , Camundongos SCID , Fagocitose , Proteínas Recombinantes , Imunodeficiência Combinada Severa/genética , Imunodeficiência Combinada Severa/imunologia , Imunodeficiência Combinada Severa/terapia , Traqueia , Transdução GenéticaRESUMO
The two mammalian neuropeptides NPFF and NPAF have been shown to have important roles in nociception, anxiety, learning and memory, and cardiovascular reflex. Two receptors (FF1 and FF2) have been molecularly identified for NPFF and NPAF. We have now characterized a novel gene designated NPVF that encodes two neuropeptides highly similar to NPFF. NPVF mRNA was detected specifically in a region between the dorsomedial and ventromedial hypothalamic nuclei. NPVF-derived peptides displayed higher affinity for FF1 than NPFF-derived peptides, but showed poor agonist activity for FF2. Following intracerebral ventricular administration, a NPVF-derived peptide blocked morphine-induced analgesia more potently than NPFF in both acute and inflammatory models of pain. In situ hybridization analysis revealed distinct expression patterns of FF1 and FF2 in the rat central nervous system. FF1 was broadly distributed, with the highest levels found in specific regions of the limbic system and the brainstem where NPVF-producing neurons were shown to project. FF2, in contrast, was mostly expressed in the spinal cord and some regions of the thalamus. These results indicate that the endogenous ligands for FF1 and FF2 are NPVF- and NPFF-derived peptides, respectively, and suggest that the NPVF/FF1 system may be an important part of endogenous anti-opioid mechanism.
Assuntos
Analgésicos/farmacologia , Morfina/farmacologia , Oligopeptídeos/isolamento & purificação , Sequência de Aminoácidos , Analgésicos/antagonistas & inibidores , Analgésicos Opioides/antagonistas & inibidores , Analgésicos Opioides/farmacologia , Animais , Células CHO , Células Cultivadas , Sistema Nervoso Central , Cricetinae , Modelos Animais de Doenças , Interações Medicamentosas , Humanos , Masculino , Camundongos , Dados de Sequência Molecular , Morfina/antagonistas & inibidores , Oligopeptídeos/genética , Oligopeptídeos/farmacologia , Dor/tratamento farmacológico , Medição da Dor/efeitos dos fármacos , Ratos , Ratos Sprague-Dawley , Homologia de Sequência de AminoácidosRESUMO
In previous studies we provided evidence that the gamma isoform of protein kinase C (PKCgamma) is an important contributor to the increased pain sensitivity that occurs after injury. Here we combined electrophysiological and behavioral approaches in wild-type and PKCgamma-null mice to compare the hyperexcitability of wide dynamic range neurons in lamina V of the spinal cord dorsal horn with the behavioral hyperexcitability produced by the same injury [application of a C-fiber irritant, mustard oil (MO), to the hindpaw]. Wild-type and null mice did not differ in their response to mechanical or thermal stimuli before tissue injury, and the magnitude of the response to the MO stimuli was comparable. In wild-type mice, MO produced a dramatic and progressive enhancement of the response of lamina V neurons to innocuous mechanical and thermal stimuli. The time course of the neuronal hyperexcitability paralleled the time course of the MO-induced behavioral allodynia (nocifensive behavior in response to a previously innocuous mechanical stimulus). Neuronal hyperexcitability was also manifest in the PKCgamma-null mice, but it lasted <30 min. By contrast, the behavioral allodynia produced by MO in the PKCgamma-null mice, although reduced to approximately half that of the wild-type mice, persisted long after the lamina V hyperexcitability had subsided. Because the MO-induced behavioral allodynia was completely blocked by an NMDA receptor antagonist, we conclude that PKCgamma mediates the transition from short- to long-term hyperexcitability of lamina V nociresponsive neurons but that the persistence of injury-induced pain must involve activity within multiple NMDA-dependent spinal cord circuits.
Assuntos
Isoenzimas/deficiência , N-Metilaspartato/metabolismo , Neuralgia/fisiopatologia , Proteína Quinase C/deficiência , Medula Espinal/fisiopatologia , Potenciais de Ação/efeitos dos fármacos , Potenciais de Ação/genética , Animais , Comportamento Animal/efeitos dos fármacos , Modelos Animais de Doenças , Antagonistas de Aminoácidos Excitatórios/farmacologia , Hiperalgesia/induzido quimicamente , Hiperalgesia/fisiopatologia , Hiperalgesia/prevenção & controle , Isoenzimas/genética , Camundongos , Camundongos Knockout , Mostardeira , Medição da Dor/efeitos dos fármacos , Estimulação Física , Extratos Vegetais/farmacologia , Óleos de Plantas , Células do Corno Posterior/fisiopatologia , Proteína Quinase C/genética , Receptores de N-Metil-D-Aspartato/antagonistas & inibidoresRESUMO
1,3-Bis(2-chloroethyl)-1-nitrosourea (BCNU) is an important cause of pulmonary toxicity. BCNU alkylates DNA at the O(6) position of guanine. O(6)-methylguanine-DNA methyltransferase (MGMT) is a DNA repair protein that removes alkyl groups from the O(6) position of guanine. To determine whether overexpression of MGMT in a lung cell reduces BCNU toxicity, the MGMT gene was transfected into A549 cells, a lung epithelial cell line. Transfected A549 cell populations demonstrated high levels of MGMT RNA, MGMT protein, and DNA repair activity. The overexpression of MGMT in lung epithelial cells provided protection from the cytotoxic effects of BCNU. Control A549 cells incubated with 100 microM BCNU had a cell survival rate of 12.5 +/- 1.2%; however, A549 cells overexpressing MGMT had a survival rate of 71.8 +/- 2.7% (P < 0.001). We also demonstrated successful transfection of MGMT into human pulmonary artery endothelial cells and a primary culture of rat type II alveolar epithelial cells with overexpression of MGMT, resulting in significant protection from BCNU toxicity. These data suggest that overexpression of DNA repair proteins such as MGMT in lung cells may protect the lung cells from cytotoxic effects of cancer chemotherapy drugs such as BCNU.
Assuntos
Carmustina/intoxicação , Pulmão/efeitos dos fármacos , O(6)-Metilguanina-DNA Metiltransferase/farmacologia , Animais , Sobrevivência Celular/efeitos dos fármacos , Células Cultivadas , Reparo do DNA , Endotélio Vascular/citologia , Endotélio Vascular/metabolismo , Humanos , Pulmão/citologia , O(6)-Metilguanina-DNA Metiltransferase/genética , Alvéolos Pulmonares/citologia , Alvéolos Pulmonares/efeitos dos fármacos , Alvéolos Pulmonares/metabolismo , Alvéolos Pulmonares/fisiologia , Artéria Pulmonar/citologia , Artéria Pulmonar/metabolismo , RNA Mensageiro/metabolismo , Ratos , TransfecçãoRESUMO
During the last 25 years, there have been substantial advances in our understanding of the physiology and pathophysiology of pain. The development of animal models that more closely mimic clinical pain in humans has helped elucidate the putative mechanisms by which chronic pain develops and is maintained. However, our increased understanding of the neurobiology of pain has not translated into breakthrough treatments for pain management. As such, chronic pain is still primarily managed by drugs whose primary indication does not include pain (eg, antidepressants, anticonvulsants, antiarrhythmics, local anesthetics). These adjuvant analgesics have come into favor despite the fact that the mechanisms through which these drugs provide pain relief remain either largely unknown or are not selective for a single target. Moreover, the efficacy of adjuvant analgesics in animal models of pain is often validated only after case studies or clinical trials have been reported. This retrospective validation of "novel" analgesics in animal models of pain raises a question of the predictive validity of these models. This article reviews the use of several adjuvant and standard analgesics currently used to treat difficult-to-manage pain. What can these drugs teach us about the development of novel pain medicines? Within this context, the use of animal models of pain to predict analgesic efficacy in clinical pain conditions is considered.
Assuntos
Macrófagos Alveolares/imunologia , Pneumonia por Pneumocystis/imunologia , Infecções Oportunistas Relacionadas com a AIDS/complicações , Infecções Oportunistas Relacionadas com a AIDS/imunologia , Animais , Linfócitos T CD4-Positivos/imunologia , Citocinas/imunologia , Infecções por HIV/complicações , Infecções por HIV/imunologia , HumanosRESUMO
Silicosis is an interstitial lung disease caused by the inhalation of crystalline silicon dioxide. Current concepts suggest that a crucial step in the development of silicosis is silica-induced injury of alveolar macrophages (AM). The adhesive protein vitronectin is a natural constituent of the lung, in which its function is largely unexplored. This study investigated a possible role for vitronectin in protecting AM from silica exposure. In this study, the concentration of vitronectin was shown to be increased in the bronchoalveolar lavage fluid of silica-treated rats. Vitronectin affinity for silica was shown both in vitro and in vivo by immunostaining. Vitronectin reduced silica-induced injury to cultured AM as determined with the (51)Cr release assay. Vitronectin reduced silica-induced free radical production as determined with a cell-free thiobarbituric acid assay. Additionally, vitronectin reduced the silica-induced respiratory burst in AM as determined with chemiluminescence. This study suggests that vitronectin may protect AM during the initial exposure to silica.
Assuntos
Macrófagos Alveolares/fisiologia , Dióxido de Silício/toxicidade , Vitronectina/fisiologia , Animais , Western Blotting , Líquido da Lavagem Broncoalveolar/química , Células Cultivadas , Radicais Livres/metabolismo , Imuno-Histoquímica , Macrófagos Alveolares/efeitos dos fármacos , Ratos , Ratos Sprague-Dawley , Explosão Respiratória/efeitos dos fármacos , Vitronectina/análise , Vitronectina/imunologiaRESUMO
The US28 gene of human cytomegalovirus (HCMV) codes a cell surface receptor for both beta chemokine and fractalkine molecules. This receptor facilitates HCMV-induced cell fusion and virus dissemination and influences susceptibility to infection with other viruses, including the human immunodeficiency virus. Five adjacent but divergent open reading frames that potentially code for molecules related to the US28 protein of HCMV are present in an African green monkey simian cytomegalovirus-derived stealth virus. This finding implies a role for chemokines in the pathogenicity of at least some stealth-adapted viruses. It may also help explain the apparent therapeutic benefit achieved in certain stealth virus-infected patients treated with agents that downregulate chemokine production.
Assuntos
Quimiocina CCL2/genética , Chlorocebus aethiops/virologia , Citomegalovirus/genética , Receptores de Quimiocinas/genética , Sequência de Aminoácidos , Animais , Clonagem Molecular , Citomegalovirus/isolamento & purificação , Infecções por Citomegalovirus/virologia , DNA Viral/análise , Síndrome de Fadiga Crônica/virologia , Humanos , Dados de Sequência Molecular , Ratos , Receptores CCR2RESUMO
Apoptosis is a form of cell death that has gained enormous attention during the past few years, and its mechanisms, important to biology and medicine, are being unraveled at an accelerating pace. Apoptosis of lung cells occurs during lung infections and may be either a host defense mechanism or reflect the pathogenesis of the infection. In the first part of this review, the biochemistry and physiology of apoptotic pathways and its regulators are discussed. This is followed by an overview of apoptotic mechanisms in selected lung infections. The implications of apoptosis in host immunity, pathogenesis, and treatment of pulmonary infections will be discussed in this context.
Assuntos
Apoptose/imunologia , Pneumonia Bacteriana/imunologia , Pneumonia Viral/imunologia , Animais , Proteína Ligante Fas , Humanos , Mediadores da Inflamação/fisiologia , Glicoproteínas de Membrana/fisiologia , Mitocôndrias/fisiologia , Alvéolos Pulmonares/fisiopatologiaRESUMO
The capsaicin (vanilloid) receptor VR1 is a cation channel expressed by primary sensory neurons of the "pain" pathway. Heterologously expressed VR1 can be activated by vanilloid compounds, protons, or heat (>43 degrees C), but whether this channel contributes to chemical or thermal sensitivity in vivo is not known. Here, we demonstrate that sensory neurons from mice lacking VR1 are severely deficient in their responses to each of these noxious stimuli. VR1-/- mice showed normal responses to noxious mechanical stimuli but exhibited no vanilloid-evoked pain behavior, were impaired in the detection of painful heat, and showed little thermal hypersensitivity in the setting of inflammation. Thus, VR1 is essential for selective modalities of pain sensation and for tissue injury-induced thermal hyperalgesia.
Assuntos
Capsaicina/farmacologia , Neurônios Aferentes/fisiologia , Nociceptores/fisiologia , Dor/fisiopatologia , Receptores de Droga/fisiologia , Animais , Temperatura Corporal/efeitos dos fármacos , Cálcio/metabolismo , Capsaicina/metabolismo , Células Cultivadas , Diterpenos/farmacologia , Gânglios Espinais/citologia , Marcação de Genes , Temperatura Alta , Concentração de Íons de Hidrogênio , Inflamação/fisiopatologia , Camundongos , Camundongos Knockout , Fibras Nervosas/fisiologia , Neurônios/fisiologia , Limiar da Dor , Medula Espinal/citologia , Medula Espinal/fisiologia , Canais de Cátion TRPVRESUMO
Silicosis is a serious occupational lung disease associated with irreversible pulmonary fibrosis. The interaction between inhaled crystalline silica and the alveolar macrophage (AM) is thought to be a key event in the development of silicosis and fibrosis. Silica can cause direct injury to AMs and can induce AMs to release various inflammatory mediators. Acute silicosis is also characterized by a marked elevation in surfactant apoprotein A (SP-A); however, the role of SP-A in silicosis is unknown. We investigated whether SP-A directly affects the response of AMs to silica. In this study, the degree of silica toxicity to cultured rat AMs as assessed by a (51)Cr cytotoxicity assay was shown to be dependent on the time of exposure and the concentration and size of the silica particles. Silica directly injured rat AMs as evidenced by a cytotoxic index of 32.9 +/- 2.5, whereas the addition of rat SP-A (5 microg/ml) significantly reduced the cytotoxic index to 16.6 +/- 1.2 (P < 0. 001). This effect was reversed when SP-A was incubated with either polyclonal rabbit anti-rat SP-A antibody or D-mannose. These data indicate that SP-A mitigates the effect of silica on AM viability, and this effect may involve the carbohydrate recognition domain of SP-A. The elevation of SP-A in acute silicosis may serve as a normal host response to prevent lung cell injury after exposure to silica.
Assuntos
Macrófagos Alveolares/efeitos dos fármacos , Proteolipídeos/farmacologia , Surfactantes Pulmonares/farmacologia , Dióxido de Silício/antagonistas & inibidores , Dióxido de Silício/intoxicação , Animais , Anticorpos/farmacologia , Sobrevivência Celular/efeitos dos fármacos , Feminino , Manose/farmacologia , Concentração Osmolar , Proteolipídeos/imunologia , Proteína A Associada a Surfactante Pulmonar , Proteínas Associadas a Surfactantes Pulmonares , Surfactantes Pulmonares/imunologia , Coelhos , Ratos , Ratos Sprague-Dawley , Fatores de TempoRESUMO
Accurate serum iron and total iron binding capacity (TIBC) measurements may be useful in acute iron overdoses. Two alumina column TIBC methods were found to measure increased TIBC when free iron was present. A homogeneous TIBC method gave consistent results until iron concentrations exceeded 500 micrograms/dL (90 mumol/L), when it began to underestimate the TIBC. Serious iron overdoses require chelation therapy with deferoxamine. Iron recovery was reduced by up to 50% for all 3 methods with clinically achievable concentrations of deferoxamine 8,400 micrograms/dL (150 mumol/L). TIBC measurements by both alumina column methods were reduced by deferoxamine in the presence of free iron and unaffected when the iron concentration was less than the TIBC. The homogeneous TIBC method yielded falsely elevated results in the presence of free deferoxamine. Procedures that measure TIBC by addition of excess ferric iron followed by alumina adsorption are not suitable for monitoring TIBC in acute iron overdose. The homogeneous TIBC assay can be used in acute iron overdose but underestimates TIBC when iron concentrations exceed 500 micrograms/dL (90 mumol/L). None of the methods examined are useful for measuring iron or TIBC in the presence of deferoxamine.