RESUMO
AIMS: CD155 is an important ligand in triggering tumour rejection by immune cells. However, the expression of CD155 and its clinical significance in hepatocellular carcinoma (HCC) remains unknown. METHODS AND RESULTS: We examined the expression level of CD155 in 174 HCC tissue samples by immunohistochemical staining and in HCC cell lines by flow cytometry; 63.8% (111 of 174) of HCC tissue samples showed negative CD155 expression. When compared with adjacent peritumour tissues, HCC tissues exhibited a significantly lower expression of CD155 (P < 0.001). Flow cytometry analysis indicated that HCC cell lines had low levels of CD155 expression. Moreover, negative CD155 expression was associated significantly with higher serum α-fetoprotein level (P = 0.016) and a higher incidence of portal vein tumour thrombus (P = 0.050). Importantly, patients with positive CD155 expression had better overall survival after surgery than those with negative CD155 expression (P = 0.005). Furthermore, Cox regression analyses showed that CD155 expression was an independent prognostic factor for HCC (P = 0.049). CONCLUSIONS: Our findings suggest that loss of CD155 expression may play an important role in the immune escape of HCC cells and thus CD155 may serve as a prognostic marker as well as a potential therapeutic target for HCC.
Assuntos
Carcinoma Hepatocelular/metabolismo , Neoplasias Hepáticas/metabolismo , Receptores Virais/metabolismo , Adulto , Idoso , Carcinoma Hepatocelular/patologia , Feminino , Citometria de Fluxo , Regulação da Expressão Gênica/genética , Humanos , Imuno-Histoquímica , Neoplasias Hepáticas/patologia , Masculino , Pessoa de Meia-Idade , Prognóstico , Células Tumorais CultivadasRESUMO
Previous studies demonstrated that melittin, the main peptide in bee venom, could cause persistent spontaneous pain, primary heat and mechanical hyperalgesia, and enhance the excitability of spinal nociceptive neurons. However, the underlying mechanism of melittin-induced cutaneous hypersensitivity is unknown. Effects of melittin applied topically to acutely dissociated rat dorsal root ganglion neurons were studied using whole-cell patch clamp and calcium imaging techniques. Melittin induced intracellular calcium increases in 60% of small (<25 µm) and medium (<40 µm) diameter sensory neurons. In current clamp, topical application of melittin evoked long-lasting firing in 55% of small and medium-sized neurons tested. In voltage clamp, melittin evoked inward currents in sensory neurons in a concentration-dependent manner. Repeated application of melittin caused increased amplitude of the inward currents. Most melittin-sensitive neurons were capsaicin-sensitive, and 65% were isolectin B4 positive. Capsazepine, the TRPV1 receptor inhibitor, completely abolished the melittin-induced inward currents and intracellular calcium transients. Inhibitions of signaling pathways showed that phospholipase A(2), but not phospholipase C, was involved in producing the melittin-induced inward currents. Inhibitors of cyclooxygenases (COX) and lipoxygenases (LOX), two key components of the arachidonic acid metabolism pathway, each partially suppressed the inward current evoked by melittin. Inhibitors of protein kinase A (PKA), but not of PKC, also abolished the melittin-induced inward currents. These results indicate that melittin can directly excite small and medium-sized sensory neurons at least in part by activating TRPV1 receptors via PLA2-COXs/LOXs cascade pathways.
Assuntos
Meliteno/farmacologia , Fosfolipases A2/metabolismo , Células Receptoras Sensoriais/efeitos dos fármacos , Canais de Cátion TRPV/agonistas , Animais , Células Cultivadas , Gânglios Espinais/citologia , Gânglios Espinais/efeitos dos fármacos , Masculino , Prostaglandina-Endoperóxido Sintases/metabolismo , Ratos , Ratos Sprague-Dawley , Células Receptoras Sensoriais/enzimologia , Células Receptoras Sensoriais/fisiologia , Canais de Cátion TRPV/antagonistas & inibidoresRESUMO
Manganese-porphyrin and -salen redox therapeutics catalyze redox reactions involving O2Ë-, H2O2, and other reactive oxygen species, thereby modulating cellular redox states. Many of these complexes perform catalase reactions via high-valent Mn-oxo or -hydroxo intermediates that oxidize H2O2 to O2, but these intermediates can also oxidize other molecules (e.g., thiols), which is peroxidase reactivity. Whether catalase or peroxidase reactivity predominates depends on the metal-ligand set and the local environment, complicating predictions of what therapeutic effects (e.g., promoting vs. suppressing apoptosis) a complex might produce in a given disease. We recently reported an organoruthenium complex (Ru1) that catalyzes ABTSË- reduction to ABTS2- with H2O2 as the terminal reductant. Given that H2O2 is thermodynamically a stronger oxidant than ABTSË-, we reasoned that the intermediate that reduced ABTSË- would also be able to reduce H2O2 to H2O. Herein we demonstrate Ru1-catalyzed H2O2 disproportionation into O2 and H2O, exhibiting an 8,580-fold faster catalase TOF vs. peroxidase TOF, which is 89.2-fold greater than the highest value reported for a Mn-porphyin or -salen complex. Furthermore, Ru1 was 120-fold more stable to H2O2 than the best MnSOD mimic (TON = 4000 vs. 33.4) Mechanistic studies provide evidence that the mechanism for Ru1-catalyzed H2O2 disproportionation is conserved with the mechanism for ABTSË- reduction. Therapeutic effects of redox catalysts can be predicted with greater accuracy for catalysts that exhibit exclusively catalase activity, thereby facilitating the development of future redox therapeutic strategies for diseases.
Assuntos
CatalaseRESUMO
Although the postsynaptic events responsible for development of pathological pain have been intensively studied, the relative contribution of presynaptic neurotransmitters to the whole process remains less elucidated. In the present investigation, we sought to measure temporal changes in spinal release of both excitatory amino acids (EAAs, glutamate and aspartate) and inhibitory amino acids (IAAs, glycine, ?-aminobutyric acid and taurine) in response to peripheral inflammatory pain state. The results showed that following peripheral chemical insult induced by subcutaneous bee venom (BV) injection, there was an initial, parallel increase in spinal release of both EAAs and IAAs, however, the balance between them was gradually disrupted when pain persisted longer, with EAAs remaining at higher level but IAAs at a level below the baseline. Moreover, the EAAs-IAAs imbalance at the spinal level was dependent upon the ongoing activity from the peripheral injury site. Intrathecal blockade of ionotropic (NMDA and non-NMDA) and metabotropic (mGluRI, II, III) glutamate receptors, respectively, resulted in a differential inhibition of BV-induced different types of pain (persistent nociception vs. hyperalgesia, or thermal vs. mechanical hyperalgesia), implicating that spinal antagonism of any specific glutamate receptor subtype fails to block all types of pain-related behaviors. This result provides a new line of evidence emphasizing an importance of restoration of EAAs-IAAs balance at the spinal level to prevent persistence or chronicity of pain.
Assuntos
Aminoácidos/metabolismo , Dor/metabolismo , Medula Espinal/metabolismo , Animais , Venenos de Abelha , Comportamento Animal/efeitos dos fármacos , Antagonistas de Aminoácidos Excitatórios/farmacologia , Aminoácidos Excitatórios/metabolismo , Temperatura Alta , Hiperalgesia/etiologia , Hiperalgesia/fisiopatologia , Inflamação/metabolismo , Inflamação/fisiopatologia , Masculino , Dor/induzido quimicamente , Dor/fisiopatologia , Limiar da Dor/efeitos dos fármacos , Estimulação Física , Ratos , Ratos Sprague-Dawley , Receptores de Glutamato/fisiologiaRESUMO
Some ruthenium-hydride complexes react with O2 to yield H2O2, therefore the principle of microscopic reversibility dictates that the reverse reaction is also possible, that H2O2 could transfer an H- to a Ru complex. Mechanistic evidence is presented, using the Ru-catalyzed ABTSË- reduction reaction as a probe, which suggests that a Ru-H intermediate is formed via deinsertion of O2 from H2O2 following coordination to Ru. This demonstration that H2O2 can function as an H- donor and reductant under biologically-relevant conditions provides the proof-of-concept that H2O2 may function as a reductant in living systems, ranging from metalloenzyme-catalyzed reactions to cellular redox homeostasis, and that H2O2 may be viable as an environmentally-friendly reductant and H- source in green catalysis.
RESUMO
A recent report from our laboratory shows that subcutaneous (s.c.) injection of melittin could induce persistent spontaneous nociception (PSN) and primary thermal or mechanical hyperalgesia. However, the exact peripheral mechanisms underlying melittin-induced multiple pain-related behaviors remain unclear. In this study, behavioral tests combined with pharmacological manipulations were used to explore potential roles of local P2X and P2Y receptors in melittin-induced inflammatory pain and hyperalgesia. Post-treatment of the primary injury site with s.c. injection of A-317491 (a potent P2X(3)/P2X(2/3) receptor antagonist) and Reactive Blue 2 (a potent P2Y receptor antagonist) could significantly suppress the development of melittin-evoked PSN and hypersensitivity (thermal and mechanical). Our control experiments demonstrated that local administration of either antagonist into the contralateral hindpaw produced no significant effect on any kind of pain-associated behaviors. Taken together, these data indicate that activation of P2X and P2Y receptors might be essential to the maintenance of melittin-induced primary thermal and mechanical hyperalgesia as well as on-going pain.
Assuntos
Meliteno/farmacologia , Nociceptores/fisiologia , Dor/fisiopatologia , Receptores Purinérgicos P2/fisiologia , Animais , Hipersensibilidade/fisiopatologia , Masculino , Fenóis/farmacologia , Compostos Policíclicos/farmacologia , Ratos , Ratos Sprague-Dawley , Receptores Purinérgicos P2XRESUMO
BACKGROUND: Extracellular signal-regulated kinase (ERK), one member of the mitogen-activated protein kinase (MAPK) family, has been suggested to regulate a diverse array of cellular functions, including cell growth, differentiation, survival, as well as neuronal plasticity. Recent evidence indicates a role for ERKs in nociceptive processing in both dorsal root ganglion and spinal cord. However, little literature has been reported to examine the differential distribution and activation of ERK isoforms, ERK1 and ERK2, at different levels of pain-related pathways under both normal and pain states. In the present study, quantitative blot immunolabeling technique was used to determine the spatial and temporal expression of ERK1 and ERK2, as well as their activated forms, in the spinal cord, primary somatosensory cortex (SI area of cortex), and hippocampus under normal, transient pain and persistent pain states. RESULTS: In naïve rats, we detected regional differences in total expression of ERK1 and ERK2 across different areas. In the spinal cord, ERK1 was expressed more abundantly than ERK2, while in the SI area of cortex and hippocampus, there was a larger amount of ERK2 than ERK1. Moreover, phosphorylated ERK2 (pERK2), not phosphorylated ERK1 (pERK1), was normally expressed with a high level in the SI area and hippocampus, but both pERK1 and pERK2 were barely detectable in normal spinal cord. Intraplantar saline or bee venom injection, mimicking transient or persistent pain respectively, can equally initiate an intense and long-lasting activation of ERKs in all three areas examined. However, isoform-dependent differences existed among these areas, that is, pERK2 exhibited stronger response than pERK1 in the spinal cord, whereas ERK1 was more remarkably activated than ERK2 in the S1 area and hippocampus. CONCLUSION: Taken these results together, we conclude that: (1) under normal state, while ERK immunoreactivity is broadly distributed in the rat central nervous system in general, the relative abundance of ERK1 and ERK2 differs greatly among specific regions; (2) under pain state, either ERK1 or ERK2 can be effectively phosphorylated with a long-term duration by both transient and persistent pain, but their response patterns differ from each other across distinct regions; (3) The long-lasting ERKs activation induced by bee venom injection is highly correlated with our previous behavioral, electrophysiological, morphological and pharmacological observations, lending further support to the functional importance of ERKs-mediated signaling pathways in the processing of negative consequences of pain associated with sensory, emotional and cognitive dimensions.
Assuntos
Encéfalo/enzimologia , Proteína Quinase 1 Ativada por Mitógeno/metabolismo , Proteína Quinase 3 Ativada por Mitógeno/metabolismo , Dor/enzimologia , Dor/patologia , Análise de Variância , Animais , Venenos de Abelha , Comportamento Animal , Encéfalo/efeitos dos fármacos , Ativação Enzimática/efeitos dos fármacos , Masculino , Dor/induzido quimicamente , Fosforilação/efeitos dos fármacos , Ratos , Ratos Sprague-Dawley , Fatores de TempoRESUMO
Our previous studies have established the idea that different types of pain induced by subcutaneous bee venom (BV) injection might be mediated by different spinal signaling pathways. To further testify this hypothesis, the present investigation was designed to detect whether spinal p38 and c-Jun N-terminal kinase (JNK) pathways are equally or differentially involved in the development of persistent spontaneous nociception (PSN), primary heat and mechanical hyperalgesia, and mirror-image heat (MIH) hypersensitivity in the BV model, by evaluating the effects of intrathecal (i.t.) pre-administration of a p38 inhibitor SB239063 and a JNK inhibitor SP600125 in the conscious rat. The results showed that i.t. pre-treatment with either SB239063 or SP600125 caused a significant prevention of BV-induced persistent paw flinching reflex in a dose-related manner, with the former exhibiting much stronger inhibition than the latter. Moreover, the same doses of SB239063 and SP600125 also exhibited different suppressive actions on the induction of primary heat hyperalgesia and MIH hypersensitivity. That is, SP600125 produced a larger increase of thermal latency than SB239063 in the injected paw, whereas SB239063 mainly affected the value measured in the non-injected paw. Pre-treatment with neither SB239063 nor SP600125 had any effect on BV-evoked mechanical hyperalgesia. Taken together, these data suggest that activation of p38 in the spinal cord preferentially contributes to the development of PSN and MIH hypersensitivity under pathological state, while spinal JNK signaling pathways might play more important roles in inducing primary heat hyperalgesia.
Assuntos
Vias Aferentes/enzimologia , Proteínas Quinases JNK Ativadas por Mitógeno/metabolismo , Nociceptores/fisiologia , Dor/enzimologia , Medula Espinal/enzimologia , Proteínas Quinases p38 Ativadas por Mitógeno/metabolismo , Vias Aferentes/efeitos dos fármacos , Animais , Venenos de Abelha/farmacologia , Relação Dose-Resposta a Droga , Ativação Enzimática/efeitos dos fármacos , Ativação Enzimática/fisiologia , Inibidores Enzimáticos/farmacologia , Hiperalgesia/induzido quimicamente , Hiperalgesia/enzimologia , Hiperalgesia/fisiopatologia , Masculino , Nociceptores/efeitos dos fármacos , Dor/induzido quimicamente , Dor/fisiopatologia , Medição da Dor/efeitos dos fármacos , Limiar da Dor/efeitos dos fármacos , Limiar da Dor/fisiologia , Ratos , Ratos Sprague-Dawley , Transdução de Sinais/efeitos dos fármacos , Transdução de Sinais/fisiologia , Medula Espinal/efeitos dos fármacosRESUMO
The precise relationship between the degree of pain and the degree of inflammation in the individual remains debated. A quantitative analysis simultaneously applied to the immediate and prolonged painful consequences of inflammation has not yet been done. Thus, the correlations between edema, nociception and hypersensitivity following an inflammatory insult were assessed in rodents. To better understand the therapeutic value of modifying specific aspects of inflammation, the effects of an anti-inflammatory drug were compared to the results. Inbred strains of mice and outbred rats received an intraplantar injection of honeybee venom and the between-group and within-group correlations were calculated for spontaneous nociceptive measures, thermal and mechanical hypersensitivity, and edema and temperature. The effect of indomethacin on the pain and inflammation measures was examined. Edema correlated with spontaneous flinching, licking and lifting of the injected paw (P< or =0.003), and not with thermal or mechanical hypersensitivity. Indomethacin affected edema and spontaneous nociception dose-dependently, and affected hypersensitivity only at the highest dose tested (P< 0.05). These results suggest that edema may contribute only to immediate spontaneous nociceptive responses to an inflammatory insult, and not to the more clinically relevant prolonged hypersensitivity. This analysis represents a method for determining which inflammatory processes are the most promising therapeutic targets against the multiple painful consequences of inflammation.
Assuntos
Anti-Inflamatórios não Esteroides/farmacologia , Edema/complicações , Indometacina/farmacologia , Inflamação/complicações , Dor/fisiopatologia , Animais , Venenos de Abelha , Edema/induzido quimicamente , Inflamação/induzido quimicamente , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Camundongos Endogâmicos C57BL , Nociceptores/fisiologia , Dor/etiologia , Medição da Dor , Ratos , Ratos Sprague-DawleyRESUMO
Carbon dots, generally small carbon nanoparticles with various forms of surface passivation, have achieved the performance level of semiconductor quantum dots in the green spectral region, but their absorption and fluorescence in red/near-IR are relatively weaker. Conceptually similar to endofullerenes, host-guest carbon dots were designed and prepared with red/near-IR dyes encapsulated as guest in the carbon nanoparticle core. Beyond the desired enhancement in optical properties, the host-guest configuration may significantly broaden the field of carbon dots.