RESUMEN
Matrix metalloproteinase 11 (MMP11), a member of the MMP family involved in the degradation of the extracellular matrix, has been implicated in cancer progression. Despite the stromal expression of MMP11 in breast cancer, the prognostic significance and role of MMP11 in immune or stromal cells of breast cancer remain unclear. Based on the immunohistochemical analysis of breast cancer tissues from 497 patients, we demonstrated that MMP11 expression in mononuclear inflammatory cells (predominantly macrophages) is an independent negative prognostic factor in breast cancer, whereas MMP11 expression in tumor cells and fibroblasts is not associated with patient survival. Enforced MMP11 expression in breast cancer cells did not promote cell proliferation and migration. However, MMP11-overexpressing macrophages enhanced the migration of HER2-positive (HER2+) breast cancer cells, recruitment of monocytes, and tube formation of endothelial cells. Furthermore, we found that the chemokine CCL2 secreted from MMP11-overexpressing macrophages activated the MAPK pathway via its receptor CCR2 in breast cancer cells, thereby promoting the migration of HER2+ breast cancer cells through MMP9 upregulation. We also found that MMP11 expression in macrophages was stimulated by MMP11-overepressing HER2+ breast cancer cells. Collectively, our findings provide evidence that MMP11 in macrophages may play a pro-tumoral role in HER2+ breast cancer through interaction with cancer cells, monocytes, and endothelial cells.
Asunto(s)
Neoplasias de la Mama , Neoplasias de la Mama/patología , Quimiocina CCL2/metabolismo , Células Endoteliales/metabolismo , Femenino , Humanos , Macrófagos/metabolismo , Metaloproteinasa 11 de la Matriz/metabolismo , Monocitos/metabolismo , Receptores CCR2RESUMEN
We explored the inhibitory effect of ginsenoside compound K (CK), 20(S)-protopanaxadiol (PPD), and 20(S)-protopanaxatriol (PPT) on six uridine 5'-diphospho-glucuronosyltransferase (UGT) enzyme (UGT1A1, 1A3, 1A4, 1A6, 1A9, and 2B7) activities in human liver microsomes (HLMs) and 10 UGT enzyme (UGT1A1, 1A3, 1A4, 1A6, 1A9, 2B4, 2B7, 2B10, 2B15, and 2B17) activities in recombinant UGT isoforms.PPD was a potent inhibitor of UGT1A3 activity with half-maximal inhibitory concentration values of 5.62 and 3.38 µM in HLMs and recombinant UGT1A3, respectively. UGT1A3 inhibition by CK and PPD was competitive with inhibitory constant (Ki) values of 17.4 and 1.21 µM, respectively, and inhibition by PPT was non-competitive with a Ki value of 8.07 µM in HLMs. PPD exhibited more than 3.4-fold selectivity for UGT1A3 inhibition compared with other UGT isoforms inhibition, while CK and PPT showed more than 2.16- and 2.21-fold selectivity, respectively.PPD did not significantly increase the mRNA expression of UGT1A1, 1A3, 1A4, 1A9, and 2B7 in hepatocytes.Given the low plasma concentrations of PPD in healthy human subjects and the absence of induction potential on UGT isoforms, we conclude that PPD cause no pharmacokinetic interactions with other co-administered drugs metabolised by UGT1A3.
Asunto(s)
Glucuronosiltransferasa , Microsomas Hepáticos , Ginsenósidos , Humanos , Sapogeninas , UridinaRESUMEN
KEY POINTS: Chronic treatment with opioids, such as morphine, leads to analgesic tolerance. While postsynaptic opioid tolerance is well documented, the involvement of presynaptic mechanisms remains unclear. We show that chronic morphine reduces the ability of periaqueductal grey (PAG) neurons to maintain GABAergic transmission. This depression of GABAergic transmission was due to a reduction in the effective size of the readily releasable pool. This also led to a reduction in opioid presynaptic inhibition; these presynaptic adaptations need to be considered in the development of strategies to reduce opioid tolerance. ABSTRACT: The midbrain periaqueductal grey (PAG) plays a critical role in tolerance to the analgesic actions of opioids such as morphine. While numerous studies have identified the postsynaptic adaptations induced by chronic morphine treatment in this and other brain regions, the presence of presynaptic adaptations remains uncertain. We examined GABAergic synaptic transmission within rat PAG brain slices from animals which underwent a low dose morphine treatment protocol which produces tolerance, but not withdrawal. Evoked GABAergic IPSCs (inhibitory postsynaptic currents) were less in morphine compared to control saline treated animals. Postsynaptic GABAA receptor mediated currents and desensitization, presynaptic release probability (Pr ), and inhibition by endogenous neurotransmitters were similar in morphine and saline treated animals. By contrast, the effective size of the readily releasable pool (RRP) was smaller in morphine treated animals. While the µ-opioid agonist DAMGO produced a reduction in Pr and RRP size in saline treated animals, it only reduced Pr in morphine treated animals. Consequently, DAMGO-induced inhibition of evoked IPSCs during short burst stimulation was less in morphine, compared to saline treated animals. These results indicate that low dose chronic morphine treatment reduces presynaptic µ-opioid inhibition by reducing the size of the pool of vesicles available for action potential dependent release. This novel presynaptic adaptation may provide important insights into the development of efficacious pain therapies that can circumvent the development of opioid tolerance.
Asunto(s)
Analgésicos Opioides/farmacología , Tolerancia a Medicamentos/fisiología , Morfina/farmacología , Sustancia Gris Periacueductal/efectos de los fármacos , Ácido gamma-Aminobutírico/metabolismo , Animales , Masculino , Neuronas/efectos de los fármacos , Neuronas/fisiología , Sustancia Gris Periacueductal/fisiología , Ratas Sprague-Dawley , Transmisión Sináptica/efectos de los fármacosRESUMEN
KEY POINTS: The midbrain periaqueductal grey (PAG) forms part of an endogenous analgesic system which is tightly regulated by the neurotransmitter GABA. The role of endocannabinoids in regulating GABAergic control of this system was examined in rat PAG slices. Under basal conditions GABAergic neurotransmission onto PAG output neurons was multivesicular. Activation of the endocannabinoid system reduced GABAergic inhibition by reducing the probability of release and by shifting release to a univesicular mode. Blockade of endocannabinoid system unmasked a tonic control over the probability and mode of GABA release. These findings provides a mechanistic foundation for the control of the PAG analgesic system by disinhibition. ABSTRACT: The midbrain periaqueductal grey (PAG) has a crucial role in coordinating endogenous analgesic responses to physiological and psychological stressors. Endocannabinoids are thought to mediate a form of stress-induced analgesia within the PAG by relieving GABAergic inhibition of output neurons, a process known as disinhibition. This disinhibition is thought to be achieved by a presynaptic reduction in GABA release probability. We examined whether other mechanisms have a role in endocannabinoid modulation of GABAergic synaptic transmission within the rat PAG. The group I mGluR agonist DHPG ((R,S)-3,5-dihydroxyphenylglycine) inhibited evoked IPSCs and increased their paired pulse ratio in normal external Ca2+ , and when release probability was reduced by lowering Ca2+ . However, the effect of DHPG on the coefficient of variation and kinetics of evoked IPSCs differed between normal and low Ca2+ . Lowering external Ca2+ had a similar effect on evoked IPSCs to that observed for DHPG in normal external Ca2+ . The low affinity GABAA receptor antagonist TPMPA ((1,2,5,6-tetrahydropyridin-4-yl)methylphosphinic acid) inhibited evoked IPSCs to a greater extent in low than in normal Ca2+ . Together these findings indicate that the normal mode of GABA release is multivesicular within the PAG, and that DHPG and lowering external Ca2+ switch this to a univesicular mode. The effects of DHPG were mediated by mGlu5 receptor engagement of the retrograde endocannabinoid system. Blockade of endocannabinoid breakdown produced a similar shift in the mode of release. We conclude that endocannabinoids control both the mode and the probability of GABA release within the PAG.
Asunto(s)
Endocannabinoides/fisiología , Sustancia Gris Periacueductal/fisiología , Sinapsis/fisiología , Ácido gamma-Aminobutírico/fisiología , Animales , Calcio/fisiología , Femenino , Antagonistas del GABA/farmacología , Técnicas In Vitro , Potenciales Postsinápticos Inhibidores/efectos de los fármacos , Masculino , Metoxihidroxifenilglicol/análogos & derivados , Metoxihidroxifenilglicol/farmacología , Sustancia Gris Periacueductal/efectos de los fármacos , Ácidos Fosfínicos/farmacología , Piperidinas/farmacología , Pirazoles/farmacología , Piridinas/farmacología , Ratas Sprague-Dawley , Receptores de Glutamato Metabotrópico/agonistas , Receptores de Glutamato Metabotrópico/fisiologíaRESUMEN
Serotonin (5-HT) modulates pain and anxiety from within the midbrain periaqueductal gray (PAG). In the present study, the effects of 5-HT- and 5-HT(1/2) subtype-selective ligands on rat PAG neurons were examined using whole cell patch-clamp recordings in brain slices. In voltage clamp, 5-HT produced outward and inward currents in distinct subpopulations of neurons that varied throughout different subregions of the PAG. The 5-HT(1A) agonist R(+)-8-OH-DPAT (1 µM) produced outward currents in subpopulations of PAG neurons. By contrast, sumatriptan (1 µM) and other 5-HT(1B, -D), and (-F) subtype agonists had little or no postsynaptic activity. The 5-HT(2A/C) agonists DOI (3 µM) and TCB-2 (1 µM) produced inward currents in subpopulations of PAG neurons, and DOI enhanced evoked inhibitory postsynaptic currents via a presynaptic mechanism. In current clamp, both R(+)-8-OH-DPAT and sumatriptan produced an excitatory increase in evoked mixed postsynaptic potentials (PSPs). In addition, R(+)-8-OH-DPAT, but not sumatriptan, directly hyperpolarized PAG neurons. By contrast, the 5-HT(2) agonist DOI depolarized subpopulations of neurons and produced an inhibitory decrease in evoked mixed PSPs. These findings indicate that 5-HT(1A) and 5-HT(1B/D) ligands have partly overlapping inhibitory effects on membrane excitability and synaptic transmission within the PAG, which are functionally opposed by 5-HT(2A/C) actions in specific PAG subregions.
Asunto(s)
Neuronas/efectos de los fármacos , Sustancia Gris Periacueductal/fisiología , Serotonina/farmacología , 8-Hidroxi-2-(di-n-propilamino)tetralin/farmacología , Animales , Potenciales Postsinápticos Inhibidores/efectos de los fármacos , Neuronas/fisiología , Sustancia Gris Periacueductal/citología , Ratas , Ratas Sprague-Dawley , Serotoninérgicos/farmacología , Sumatriptán/farmacologíaRESUMEN
MicroRNA (miRNA) is a class of endogenous small noncoding RNA that negatively regulates gene expression at the post-transcriptional level and plays an important role in the pathogenesis of various diseases. However, the identity and role of miRNAs involved in the development of insulin resistance resulting from mitochondrial dysfunction are largely unknown. In this study, mitochondrial dysfunction by genetic or metabolic inhibition induced an impairment of insulin signaling in SK-Hep1 cells via a reduction in the expression of IRS-1 protein. Significant up-regulation of miR-96, which was presumed to target IRS-1 3'UTR, was found in SK-Hep1 cells with mitochondrial dysfunction. Using reporter gene assay we confirmed that miR-96 authentically targeted IRS-1 3'UTR. Furthermore, the ectopic expression of miR-96 caused a substantial decrease in IRS-1 protein expression, and a consequent impairment in insulin signaling. These findings suggest that the up-regulation of miR-96 by mitochondrial dysfunction contributes to the development of insulin resistance by targeting IRS-1 in SK-Hep1 cells.
Asunto(s)
Glucógeno/biosíntesis , Proteínas Sustrato del Receptor de Insulina/genética , MicroARNs/biosíntesis , Mitocondrias/fisiología , Biosíntesis de Proteínas , Regiones no Traducidas 3' , Línea Celular Tumoral , Electroforesis en Gel de Poliacrilamida , Humanos , Insulina/metabolismo , Transducción de Señal , Regulación hacia ArribaRESUMEN
Introduction: This study investigates attention mechanisms and the accuracy of emotion judgment among South Korean children by employing Korean and American faces in conjunction with eye-tracking technology. Methods: A total of 42 participants were individually presented with photos featuring either Korean or American children, and their task was to judge the emotions conveyed through the facial expressions in each photo. The participants' eye movements during picture viewing were meticulously observed using an eye tracker. Results: The analysis of the emotion judgment task outcomes revealed that the accuracy scores for discerning emotions of joy, sadness, and anger in Korean emotional faces were found to be significantly higher than those for American children. Conversely, no significant difference in accuracy scores was observed for the recognition of fear emotion between Korean and American faces. Notably, the study also uncovered distinct patterns of fixation duration among children, depending on whether they were viewing Korean or American faces. These patterns predominantly manifested in the three main facial areas of interest, namely the eyes, nose, and mouth. Discussion: The observed phenomena can be best understood within the framework of the "other-race effect." Consequently, this prototype formation leads to heightened accuracy in recognizing and interpreting emotional expressions exhibited by faces belonging to the same racial group. The present study contributes to a deeper understanding of how attention mechanisms and other-race effects impact emotion judgment among South Korean children. The utilization of eye-tracking technology enhances the validity and precision of our findings, providing valuable insights for both theoretical models of face processing and practical applications in various fields such as psychology, education, and intercultural communication.
RESUMEN
In the concrete industry, various R&D efforts have been devoted to self-healing technology, which can maintain the long-term performance of concrete structures, which is important in terms of sustainable development. Cracks in cement composites occur and propagate because of various internal and external factors, reducing the composite's stability. Interest in "self-healing" materials that can repair cracks has led researchers to embed self-healing capsules in cement composites. Overcoming the limitations of polymer capsules produced by chemical manufacturing methods, three-dimensional (3D) printing can produce capsules quickly and accurately and offers advantages such as high material strength, low cost, and the ability to fabricate capsules with complex geometries. We performed structural analysis simulations, experimentally evaluated the mechanical properties and solubility of poly(lactic acid) (PLA) capsules, and examined the effect of the capsule wall thickness and printing direction on cement composites embedded with these capsules. Thicker capsules withstood larger bursting loads, and the capsule rupture characteristics varied with the printing angle. Thus, the capsule design parameters must be optimized for different environments. Although the embedded capsules slightly reduced the compressive strength of the cement composites, the benefit of the encapsulated self-healing agent is expected to overcome this disadvantage.
RESUMEN
BACKGROUND: There is anatomical and behavioural evidence that µ- and δ-opioid receptors modulate distinct nociceptive modalities within the superficial dorsal horn. The aim of the present study was to examine whether µ- and δ-opioid receptor activation differentially modulates TRP sensitive inputs to neurons within the superficial dorsal horn. To do this, whole cell patch clamp recordings were made from lamina I - II neurons in rat spinal cord slices in vitro to examine the effect of opioids on TRP agonist-enhanced glutamatergic spontaneous miniature excitatory postsynaptic currents (EPSCs). RESULTS: Under basal conditions the µ-opioid agonist DAMGO (3 µM) reduced the rate of miniature EPSCs in 68% of neurons, while the δ- and κ-opioid agonists deltorphin-II (300 nM) and U69593 (300 nM) did so in 13 - 17% of neurons tested. The TRP agonists menthol (400 µM) and icilin (100 µM) both produced a Ca2+-dependent increase in miniature EPSC rate which was unaffected by the voltage dependent calcium channel (VDCC) blocker Cd2+. The proportion of neurons in which deltorphin-II reduced the miniature EPSC rate was enhanced in the presence of icilin (83%), but not menthol (0%). By contrast, the proportion of DAMGO and U69593 responders was unaltered in the presence of menthol (57%, 0%), or icilin (57%, 17%). CONCLUSIONS: These findings demonstrate that δ-opioid receptor activation selectively inhibits inputs activated by icilin, whereas µ-opioid receptor activation has a more widespread effect on synaptic inputs to neurons in the superficial dorsal horn. These findings suggest that δ-opioids may provide a novel analgesic approach for specific, TRPA1-like mediated pain modalities.
Asunto(s)
Glutamatos/metabolismo , Células del Asta Posterior/metabolismo , Receptores Opioides delta/metabolismo , Receptores Opioides mu/metabolismo , Transmisión Sináptica , Animales , Calcio/metabolismo , Canales de Calcio/metabolismo , Potenciales Postsinápticos Excitadores/efectos de los fármacos , Técnicas In Vitro , Masculino , Mentol/farmacología , Células del Asta Posterior/efectos de los fármacos , Terminales Presinápticos/efectos de los fármacos , Terminales Presinápticos/metabolismo , Pirimidinonas/farmacología , Ratas , Ratas Sprague-Dawley , Transmisión Sináptica/efectos de los fármacosRESUMEN
Acrylamide, a known carcinogen, is formed during preparation of food containing reducing sugar and asparagine. Because acrylamide exposure of the population is primarily through food, the maximum levels of acrylamide in food were set by the European Commission in 2017. Moreover, in the 2016 Korean Total Diet study, acrylamide showed the lowest margin of exposure among 23 food-processing-related chemicals, necessitating risk reduction options. Therefore, the objective of this study was to determine the variation of acrylamide content in different food items and identify the food categories, to provide options for risk management. Acrylamide was analysed using high-performance liquid chromatography coupled with tandem mass spectrometry in more than 1,000 processed food items. To estimate acrylamide exposure, the analytical data obtained herein and the food consumption data of Korean National Health and Nutrition Examination Survey from 2013 to 2017 were used. A non-parametric technique of a probabilistic model was used for exposure estimation. Confectioneries (here this category includes potato and similar savoury snacks) contained a wide range of acrylamide content. Particularly, the highest acrylamide content was detected in a tea made of Jerusalem artichoke. The presence of acrylamide in turmeric along with the Jerusalem artichoke was reported in this study for the first time. The main contributors of dietary acrylamide exposure were confectioneries for youths aged 3-18 years and coffee for adults aged 19-80 years. Therefore, risk management in confectionery and coffee could help reduce acrylamide exposure for Koreans. In addition, the mitigation strategies for food containing high acrylamide content, such as Jerusalem artichoke tea, are needed to reduce acrylamide exposure to loyal consumers.
Asunto(s)
Acrilamida/análisis , Exposición Dietética/análisis , Análisis de los Alimentos , Contaminación de Alimentos/análisis , Modelos Estadísticos , Encuestas Nutricionales , Adolescente , Adulto , Anciano , Anciano de 80 o más Años , Niño , Preescolar , Femenino , Humanos , Masculino , Persona de Mediana Edad , República de Corea , Estadísticas no Paramétricas , Adulto JovenRESUMEN
BACKGROUND AND PURPOSE: While triptans are used to treat migraine, there is evidence that they also reduce inflammation-induced pain at the spinal level. The cellular mechanisms underlying this spinal enhancement are unknown. We examined whether inflammation alters sumatriptan modulation of synaptic transmission in the rat spinal dorsal horn. EXPERIMENTAL APPROACH: Three to four days following intraplantar injection of complete Freund's adjuvant (CFA) or saline, whole cell recordings of evoked glutamatergic EPSCs were made from lumbar lamina I-II dorsal horn neurons in rat spinal slices KEY RESULTS: In 2- to 3-week-old animals, sumatriptan reduced the amplitude of evoked EPSCs and this was greater in slices from CFA, compared to saline-injected rats. In CFA-injected animals, sumatriptan increased the paired pulse ratio of evoked EPSCs and reduced the rate of spontaneous miniature EPSCs. The 5-HT1B and 5-HT1D agonists CP9 3129 and PNU109291 both inhibited evoked EPSCs in CFA but not saline-injected rats. By contrast, the 5-HT1A agonist R(+)-8-OH-DPAT inhibited evoked EPSCs in saline but not CFA-injected rats. In CFA-injected rats, the sumatriptan-induced inhibition of evoked EPSCs was reduced by the 5-HT1B and 5-HT1D antagonists NAS181 and BRL-15572. Intriguingly, the difference in sumatriptan inhibition between CFA and saline-injected animals was only observed in animals less than 4 weeks old. CONCLUSION AND IMPLICATIONS: These findings indicate that inflammation induces a developmentally regulated 5-HT1B/1D presynaptic inhibition of excitatory transmission into the rat superficial dorsal horn. Thus, triptans could potentially act as spinal analgesic agents for inflammatory pain in the juvenile setting.
Asunto(s)
Sumatriptán , Transmisión Sináptica , Animales , Inflamación/inducido químicamente , Inflamación/tratamiento farmacológico , Células del Asta Posterior , Ratas , Ratas Sprague-Dawley , Médula Espinal , Sumatriptán/farmacologíaRESUMEN
A chemo- and site-selective reduction reaction of isatin derivatives using catalyst B(C6F5)3 and hydrosilanes is described. This transformation is operationally simple, proceeds under mild conditions, and is resistant to various functional groups. Thus, this efficient reaction using a combination of B(C6F5)3 and BnMe2SiH or B(C6F5)3 and Et2SiH2 could potentially be utilized to produce various indolin-3-ones and indolines, without the need for multistep procedures and metal catalysis conditions.
RESUMEN
BACKGROUND: There is evidence to suggest that the midbrain periaqueductal grey (PAG) has a role in migraine and the actions of the anti-migraine drug sumatriptan. In the present study we examined the serotonergic modulation of GABAergic and glutamatergic synaptic transmission in rat midbrain PAG slices in vitro. RESULTS: Serotonin (5-hydroxytriptamine, 5-HT, IC50 = 142 nM) and the selective serotonin reuptake inhibitor fluoxetine (30 microM) produced a reduction in the amplitude of GABAA-mediated evoked inhibitory postsynaptic currents (IPSCs) in all PAG neurons which was associated with an increase in the paired-pulse ratio of evoked IPSCs. Real time PCR revealed that 5-HT1A, 5-HT1B, 5-HT1D and 5-HT1F receptor mRNA was present in the PAG. The 5-HT1A, 5-HT1B and 5-HT1D receptor agonists 8-OH-DPAT (3 microM), CP93129 (3 microM) and L694247 (3 microM), but not the 5-HT1F receptor agonist LY344864 (1 - 3 microM) inhibited evoked IPSCs. The 5-HT (1 microM) induced inhibition of evoked IPSCs was abolished by the 5-HT1B antagonist NAS181 (10 microM), but not by the 5-HT1A and 5-HT1D antagonists WAY100135 (3 microM) and BRL15572 (10 microM). Sumatriptan also inhibited evoked IPSCs with an IC50 of 261 nM, and reduced the rate, but not the amplitude of spontaneous miniature IPSCs. The sumatriptan (1 microM) induced inhibition of evoked IPSCs was abolished by NAS181 (10 microM) and BRL15572 (10 microM), together, but not separately. 5-HT (10 microM) and sumatriptan (3 microM) also reduced the amplitude of non-NMDA mediated evoked excitatory postsynaptic currents (EPSCs) in all PAG neurons tested. CONCLUSION: These results indicate that sumatriptan inhibits GABAergic and glutamatergic synaptic transmission within the PAG via a 5-HT1B/D receptor mediated reduction in the probability of neurotransmitter release from nerve terminals. These actions overlap those of other analgesics, such as opioids, and provide a mechanism by which centrally acting 5-HT1B and 5-HT1D ligands might lead to novel anti-migraine pharmacotherapies.
Asunto(s)
Sustancia Gris Periacueductal/fisiología , Agonistas de Receptores de Serotonina/farmacología , Sumatriptán/farmacología , Transmisión Sináptica/efectos de los fármacos , Animales , Ácido Glutámico , Mesencéfalo/fisiología , Ratas , Receptor de Serotonina 5-HT1B , Receptor de Serotonina 5-HT1D , Ácido gamma-AminobutíricoRESUMEN
Molecular biology approaches have identified more than 70 different K+ channel genes that assemble to form diverse functional classes of K+ channels. Although functional K+ channels are present within presynaptic nerve endings, direct studies of their precise identity and function have been generally limited to large, specialized presynaptic terminals such as basket cell terminals and Calyx of Held. In the present study, therefore, we investigated the functional K+ channel subtypes on the small glycinergic nerve endings (< 1 microm diameter) projecting to spinal sacral dorsal commissural nucleus (SDCN) neurons. In the presence of TTX, whole-cell patch recording of mIPSCs was made from mechanically dispersed SDCN neurons in which functional nerve endings remain attached. Glycinergic responses were isolated by blocking glutamatergic and GABAergic inputs with CNQX, AP5 and bicuculline. The K+ channel blockers, 4-AP, TEA, delta-dendrotoxin, margatoxin, iberiotoxin, charybdotoxin and apamin, significantly increased 'spontaneous' mIPSC frequency without affecting mIPSC amplitude. The results suggest the existence of the following K+ channel subtypes on glycinergic nerve endings that are involved in regulating 'spontaneous' glycine release (mIPSCs): the Shaker-related K+ channels Kv1.1, Kv1.2, Kv1.3, Kv1.6 and Kv1.7 and the intracellular Ca2+ -sensitive K+ channels BKCa, IKCa and SKCa. Ca2+ channel blockers by themselves, including L-type (nifedipine), P/Q-type (omega-agatoxin IVA, AgTX) and N-type (omega-conotoxin GVIA, CgTX), did not alter the 'spontaneous' mIPSC frequency or amplitude, but inhibited the increase of the mIPSC frequency evoked by 4-AP, indicating the participation of L-, P/Q- and N-type Ca2+ channels regulating 'spontaneous' glycine release from the nerve terminals.
Asunto(s)
Glicina/metabolismo , Inhibición Neural/fisiología , Neuronas/metabolismo , Bloqueadores de los Canales de Potasio/farmacología , Canales de Potasio/metabolismo , Médula Espinal/metabolismo , Animales , Bloqueadores de los Canales de Calcio/farmacología , Canales de Calcio/efectos de los fármacos , Canales de Calcio/metabolismo , Potenciales Postsinápticos Inhibidores/efectos de los fármacos , Potenciales Postsinápticos Inhibidores/fisiología , Inhibición Neural/efectos de los fármacos , Vías Nerviosas/efectos de los fármacos , Vías Nerviosas/metabolismo , Neuronas/efectos de los fármacos , Técnicas de Cultivo de Órganos , Técnicas de Placa-Clamp , Canales de Potasio/efectos de los fármacos , Canales de Potasio Calcio-Activados/efectos de los fármacos , Canales de Potasio Calcio-Activados/metabolismo , Canales de Potasio con Entrada de Voltaje/efectos de los fármacos , Canales de Potasio con Entrada de Voltaje/metabolismo , Terminales Presinápticos/efectos de los fármacos , Terminales Presinápticos/metabolismo , Ratas , Ratas Wistar , Médula Espinal/citología , Médula Espinal/efectos de los fármacos , Transmisión Sináptica/efectos de los fármacos , Transmisión Sináptica/fisiologíaRESUMEN
SCOPE: Obesity increases intracellular lipid accumulation in key tissues or organs, which often leads to metabolic dysfunction and insulin resistance. Diets rich in saturated fatty acid (SFA) exacerbate obesity and hepatic steatosis, which accentuate the risk of insulin resistance and type 2 diabetes (T2DM). Although microRNAs (miRNAs) play a critical role in the regulation of gene expression, the implication of obesity-induced miRNAs in metabolic disorders particularly in the development of insulin resistance is largely unknown. Here, we investigated the implication of miR-15b, which is induced by SFA palmitate or obesity, in hepatic insulin resistance. METHODS AND RESULTS: Diet-induced obesity (DIO) in mice developed hyperglycemia and insulin resistance, accompanying with a reduction of insulin receptor (INSR) expression. Palmitate impaired insulin signaling as well as a decrease of INSR in hepatocytes. The expression of miR-15b was upregulated by DIO or palmitate in hepatocytes. Furthermore, the overexpression of miR-15b suppressed the protein expression of INSR through targeting INSR 3' untranslated region directly, resulting in an impairment of the insulin signaling and glycogen synthesis in hepatocytes. CONCLUSION: These results unveil a novel mechanism whereby miR-15b is linked causally to the pathogenesis of hepatic insulin resistance in SFA-induced obesity.
Asunto(s)
Hepatocitos/metabolismo , Resistencia a la Insulina , MicroARNs/fisiología , Obesidad/metabolismo , Receptor de Insulina/fisiología , Animales , Dieta Alta en Grasa , Células Hep G2 , Humanos , Hígado/metabolismo , Masculino , Ratones , Ratones Endogámicos C57BL , Palmitatos/farmacología , Receptor de Insulina/análisis , Receptor de Insulina/genéticaRESUMEN
MicroRNAs (miRNAs) play an important role in insulin signaling and insulin secretion, but the role of miRNAs in the association between obesity and hepatic insulin resistance is largely unknown. This study reports that saturated fatty acid (SFA) and high fat diet (HFD) significantly induce miR-195 expression in hepatocytes, and that the insulin receptor (INSR), not insulin receptor substrate-1 (IRS-1), is a direct target of miR-195. Furthermore, the ectopic expression of miR-195 suppresses the expression of INSR, thereby impairing the insulin signaling cascade and glycogen synthesis in HepG2 cells. These findings suggest that the dysregulation of miR-195 by SFA is a detrimental factor for hepatic insulin sensitivity.
Asunto(s)
Ácidos Grasos/farmacología , Glucógeno/metabolismo , Insulina/metabolismo , MicroARNs/genética , Transducción de Señal/efectos de los fármacos , Regiones no Traducidas 3'/genética , Animales , Sitios de Unión , Dieta Alta en Grasa/efectos adversos , Células Hep G2 , Hepatocitos/citología , Hepatocitos/efectos de los fármacos , Hepatocitos/metabolismo , Hepatocitos/patología , Humanos , Resistencia a la Insulina , Ratones , Obesidad/metabolismo , Obesidad/patología , Receptor de Insulina/genética , Regulación hacia Arriba/efectos de los fármacosRESUMEN
MicroRNAs have been shown to play an important role in insulin signaling but their biological function in insulin resistance induced by saturated fatty acids (SFA) remains largely unknown. Here, we report that SFA palmitate and high fat diet (HFD) significantly increase expression of miR-29a in myocytes. miR-29a targets IRS-1 3'UTR directly and represses IRS-1 expression at the translational level. Furthermore, the ectopic expression of miR-29a impairs insulin signaling and glucose uptake in myocytes through a substantial decrease in IRS-1. These findings suggest that the up-regulation of miR-29a by SFA is causally related to the development of insulin resistance in myocytes.
Asunto(s)
Ácidos Grasos/farmacología , Glucosa/metabolismo , Proteínas Sustrato del Receptor de Insulina/genética , Insulina/metabolismo , MicroARNs/biosíntesis , MicroARNs/genética , Fibras Musculares Esqueléticas/efectos de los fármacos , Fibras Musculares Esqueléticas/metabolismo , Regiones no Traducidas 3' , Animales , Secuencia de Bases , Línea Celular , Dieta Alta en Grasa , Ácidos Grasos/metabolismo , Resistencia a la Insulina , Ratones , Ratones Endogámicos C57BL , Ácido Palmítico/metabolismo , Ácido Palmítico/farmacología , Biosíntesis de Proteínas/efectos de los fármacos , Transducción de Señal/efectos de los fármacos , Regulación hacia Arriba/efectos de los fármacosRESUMEN
Diabetes is a complex metabolic disease with an etiology that includes genetic, epigenetic and environmental factors that lead to several different defects of glucose homeostasis, primarily in the pancreatic ß-cells, liver, muscle, and adipose tissues. MicroRNAs (miRNAs) have recently emerged as important regulators in post-transcriptional gene expression. Although the target genes and biological functions of individual miRNAs remain largely unknown, previous studies have shown them to be important regulators of diverse biological processes, in both normal and pathological states. In the past decade, an increasing number of studies have focused on the regulatory roles of miRNAs in metabolism; thus, miRNAs play an important role in the pathogenesis of diabetes. This review summarizes recent findings related to the roles of miRNAs in diabetes. The information presented herein might be useful for the future development of miRNAs as diagnostic and therapeutic targets in diabetes.
Asunto(s)
Diabetes Mellitus/etiología , Diabetes Mellitus/genética , Terapia Genética/tendencias , Enfermedades Metabólicas/genética , Enfermedades Metabólicas/metabolismo , MicroARNs/fisiología , Animales , Diabetes Mellitus/terapia , Silenciador del Gen/fisiología , Humanos , Enfermedades Metabólicas/terapia , MicroARNs/antagonistas & inhibidoresRESUMEN
BACKGROUND AND PURPOSE: 5-HT receptor agonists have variable nociceptive effects within the spinal cord. While there is some evidence for 5-HT(1A) spinally-mediated analgesia, the role of other 5-HT(1) receptor subtypes remains unclear. In the present study, we examined the spinal actions of a range of 5-HT(1) agonists, including sumatriptan, on acute pain, plus their effect on afferent-evoked synaptic transmission onto superficial dorsal horn neurons. EXPERIMENTAL APPROACH: For in vivo experiments, 5-HT agonists were injected via chronically implanted spinal catheters to examine their effects in acute mechanical and thermal pain assays using a paw pressure analgesymeter and a Hargreave's device. For in vitro experiments, whole-cell patch-clamp recordings of primary afferent-evoked glutamatergic EPSC were made from lamina II neurons in rat lumbar spinal slices. KEY RESULTS: Intrathecal (i.t.) delivery of the 5-HT(1A) agonist R ± 8-OH-DPAT (30-300 nmol) produced a dose-dependent thermal, but not mechanical, analgesia. Sumatriptan and the 5-HT(1B), 5-HT(1D), 5-HT(1F) agonists CP93129, PNU109291 and LY344864 (100 nmol) had no effect on either acute pain assay. R ± 8-OH-DPAT (1 µM) and sumatriptan (3 µM) both reduced the amplitude of the evoked EPSC. In contrast, CP93129, PNU109291 and LY344864 (0.3-3 µM) had no effect on the evoked EPSC. The actions of both R ± 8-OH-DPAT and sumatriptan were abolished by the 5-HT(1A) antagonist WAY100635 (3 µM). CONCLUSIONS AND IMPLICATIONS: These findings indicate that the 5-HT(1A) receptor subtype predominantly mediates the acute antinociceptive and cellular actions of 5-HT(1) ligands within the rat superficial dorsal horn.
Asunto(s)
Dolor Agudo/fisiopatología , Células del Asta Posterior/fisiología , Receptores de Serotonina 5-HT1/fisiología , Transmisión Sináptica/fisiología , Dolor Agudo/tratamiento farmacológico , Analgésicos/farmacología , Analgésicos/uso terapéutico , Animales , Células del Asta Posterior/efectos de los fármacos , Ratas , Ratas Sprague-Dawley , Agonistas de Receptores de Serotonina/farmacología , Agonistas de Receptores de Serotonina/uso terapéutico , Transmisión Sináptica/efectos de los fármacosRESUMEN
Cholecystokinin modulates pain and anxiety via its functions within brain regions such as the midbrain periaqueductal gray (PAG). The aim of this study was to examine the cellular actions of cholecystokinin on PAG neurons. Whole-cell patch clamp recordings were made from rat midbrain PAG slices in vitro to examine the postsynaptic effects of cholecystokinin and its effects on synaptic transmission. Sulfated cholecystokinin-(26-33) (CCK-S, 100-300 nM), but not non-sulfated cholecystokinin-(26-33) (CCK-NS, 100-300 nM) produced an inward current in a sub-population of opioid sensitive and insensitive PAG neurons, which did not reverse over a range of membrane potentials. The CCK-S-induced current was abolished by the CCK1 selective antagonist devazepide (100 nM), but not by the CCK2 selective antagonists CI988 (100 nM, 1 µM) and LY225910 (1 µM). CCK-S, but not CCK-NS produced a reduction in the amplitude of evoked GABA(A)-mediated inhibitory postsynaptic currents (IPSCs) and an increase in the evoked IPSC paired-pulse ratio. By contrast, CCK-S had little effect on the rate and amplitude of TTX-resistant miniature IPSCs under basal conditions and when external K(+) was elevated. The CCK-S-induced inhibition of evoked IPSCs was abolished by the cannabinoid CB1 receptor antagonist AM251 (3 µM), the mGluR5 antagonist MPEP (10 µM) and the 1, 2-diacylglycerol lipase (DAGLα) inhibitor tetrahydrolipstatin (10 µM). In addition, CCK-S produced an increase in the rate of spontaneous non-NMDA-mediated, TTX-dependent excitatory postsynaptic currents (EPSCs). These results suggest that cholecystokinin produces direct neuronal depolarisation via CCK1 receptors and inhibits GABAergic synaptic transmission via action potential-dependent release of glutamate and mGluR5-induced endocannabinoid signaling. Thus, cholecystokinin has cellular actions within the PAG that can both oppose and reinforce opioid and cannabinoid modulation of pain and anxiety within this brain structure.