RESUMEN
BACKGROUND: Synapses can adapt to changes in the intracerebral microenvironment by regulation of presynaptic neurotransmitter release and postsynaptic neurotransmitter receptor expression following hypoxic ischemia (HI) injury. The peptide neurotransmitter N-acetylaspartylglutamate (NAAG) exerts a protective effect on neurons after HI and may be involved in maintaining the function of synaptic networks. In this study, we investigated the changes in the expression of NAAG, glutamic acid (Glu) and metabotropic glutamate receptors (mGluRs), as well as the dynamic regulation of neurotransmitters in the brain after HI, and assessed their effects on synaptic plasticity of the cerebral cortex. METHODS: Thirty-six Yorkshire newborn pigs (3-day-old, males, 1.0-1.5 kg) were selected and randomly divided into normal saline (NS) group (n = 18) and glutamate carboxypeptidase II inhibition group (n = 18), both groups were divided into control group, 0-6 h, 6-12 h, 12-24 h, 24-48 h and 48-72 h groups (all n = 3) according to different post-HI time. The content of Glu and NAAG after HI injury were detected by 1H-MRS scanning, immunofluorescence staining of mGluRs, synaptophysin (syph) along with postsynaptic density protein-95 (PSD95) and transmission electron microscopy were performed. ANOVA, Tukey and LSD test were used to compare the differences in metabolite and protein expression levels among subgroups. Correlation analysis was performed using Pearson analysis with a significance level of α = 0.05. RESULTS: We observed that the NAAG and mGluR3 expression levels in the brain increased and then decreased after HI and was significantly higher in the 12-24 h (P < 0.05, Tukey test). There was a significant positive correlation between Glu content and the expression of mGluR1/mGluR5 after HI with r = 0.521 (P = 0.027) and r = 0.477 (P = 0.045), respectively. NAAG content was significantly and positively correlated with the level of mGluR3 expression (r = 0.472, P = 0.048). When hydrolysis of NAAG was inhibited, the expression of synaptic protein PSD95 and syph decreased significantly. CONCLUSIONS: After 12-24 h of HI injury, there was a one-time elevation in NAAG levels, which was consistent with the corresponding mGluR3 receptor expression trend; the NAAG maintains cortical synaptic plasticity and neurotransmitter homeostasis by inhibiting presynaptic glutamate vesicle release, regulating postsynaptic density proteins and postsynaptic receptor expression after pathway activation. Video abstract.
Asunto(s)
Dipéptidos , Receptores de Glutamato Metabotrópico , Animales , Corteza Cerebral/metabolismo , Dipéptidos/metabolismo , Dipéptidos/farmacología , Ácido Glutámico , Isquemia , Masculino , Plasticidad Neuronal/fisiología , Neurotransmisores/farmacología , Receptores de Glutamato Metabotrópico/metabolismo , PorcinosRESUMEN
PURPOSE: We aimed to evaluate the reproducibility of computed tomography (CT) radiomic features (RFs) about Epicardial Adipose Tissue (EAT). The features derived from coronary photon-counting computed tomography (PCCT) angiography datasets using the PureCalcium (VNCPC) and conventional virtual non-contrast (VNCConv) algorithm were compared with true non-contrast (TNC) series. METHODS: RFs of EAT from 52 patients who underwent PCCT were quantified using VNCPC, VNCConv, and TNC series. The agreement of EAT volume (EATV) and EAT density (EATD) was evaluated using Pearson's correlation coefficient and Bland-Altman analysis. A total of 1530 RFs were included. They are divided into 17 feature categories, each containing 90 RFs. The intraclass correlation coefficients (ICCs) and concordance correlation coefficients (CCCs) were calculated to assess the reproducibility of RFs. The cutoff value considered indicative of reproducible features was > 0.75. RESULTS: the VNCPC and VNCConv tended to underestimate EATVs and overestimate EATDs. Both EATV and EATD of VNCPC series showed higher correlation and agreement with TNC than VNCConv series. All types of RFs from VNCPC series showed greater reproducibility than VNCConv series. Across all image filters, the Square filter exhibited the highest level of reproducibility (ICC = 67/90, 74.4%; CCC = 67/90, 74.4%). GLDM_GrayLevelNonUniformity feature had the highest reproducibility in the original image (ICC = 0.957, CCC = 0.958), exhibiting a high degree of reproducibility across all image filters. CONCLUSION: The accuracy evaluation of EATV and EATD and the reproducibility of RFs from VNCPC series make it an excellent substitute for TNC series exceeding VNCConv series.
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Tejido Adiposo , Algoritmos , Angiografía por Tomografía Computarizada , Angiografía Coronaria , Enfermedad de la Arteria Coronaria , Pericardio , Valor Predictivo de las Pruebas , Interpretación de Imagen Radiográfica Asistida por Computador , Humanos , Reproducibilidad de los Resultados , Tejido Adiposo/diagnóstico por imagen , Pericardio/diagnóstico por imagen , Femenino , Masculino , Persona de Mediana Edad , Anciano , Angiografía Coronaria/métodos , Enfermedad de la Arteria Coronaria/diagnóstico por imagen , Estudios Retrospectivos , Vasos Coronarios/diagnóstico por imagen , Tomografía Computarizada Multidetector , Adiposidad , Tejido Adiposo Epicárdico , RadiómicaRESUMEN
Following neonatal hypoxic-ischemia (HI) injury, it is crucial factor to reconstruct neural circuit and maintain neural network homeostasis for neurological recovery. A dynamic balance between the synthesis and degradation of synaptic protein is required for maintaining synaptic plasticity. Protein degradation is facilitated by autophagy. This study aimed to investigate the regulation of synaptic structural plasticity by the Notch pathway, by assessing changes in Notch pathway activation and their effects on synaptic proteins and autophagy after HI injury. The study involved 48 male newborn Yorkshire piglets, each weighing 1.0-1.5 kg and 3 days old. They were randomly assigned to two groups: the HI group and the Notch pathway inhibitor + HI group (n = 24 per group). Each group was further divided into six subgroups according to HI duration (n = 4 per group): a control subgroup, and 0-6, 6-12, 12-24, 24-48, and 48-72 h subgroups. The expression of Notch pathway-related proteins, including Notch1, Hes1, and Notch intracellular domains, increased following HI injury. The expression of autophagy proteins increased at 0-6 h and 6-12 h post-HI. The expression of synaptic proteins, such as postsynaptic density protein 95 (PSD95) and synaptophysin, increased 6-12 h and 12-24 h after HI, respectively. Notably, the increased expression of these proteins was reversed by a Notch pathway inhibitor. Transmission electron microscopy revealed the presence of autophagosome structures in synapses. These findings shed light on the underlying mechanisms of neurological recovery after HI injury and may provide insights into potential therapeutic targets for promoting neural circuit reconstruction and maintaining neural network homeostasis.
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Hipoxia-Isquemia Encefálica , Porcinos , Animales , Masculino , Animales Recién Nacidos , Hipoxia-Isquemia Encefálica/tratamiento farmacológico , Hipoxia-Isquemia Encefálica/metabolismo , Hipoxia , Isquemia , Autofagia , Plasticidad NeuronalRESUMEN
Maintaining glutamate homeostasis after hypoxic ischemia is important for synaptic function and neural cell activity, and regulation of glutamate transport between astrocyte and neuron is one of the important modalities for reducing glutamate accumulation. However, further research is needed to investigate the dynamic changes in and molecular mechanisms of glutamate transport and the effects of glutamate transport on synapses. The aim of this study was to investigate the regulatory mechanisms underlying Notch pathway mediation of glutamate transport and synaptic plasticity. In this study, Yorkshire neonatal pigs (male, age 3 days, weight 1.0-1.5 kg, n = 48) were randomly divided into control (sham surgery group) and five hypoxic ischemia subgroups, according to different recovery time, which were then further subdivided into subgroups treated with dimethyl sulfoxide or a Notch pathway inhibitor (N-[N-(3, 5-difluorophenacetyl-l-alanyl)]-S-phenylglycine t-butyl ester). Once the model was established, immunohistochemistry, immunofluorescence staining, and western blot analyses of Notch pathway-related proteins, synaptophysin, and glutamate transporter were performed. Moreover, synapse microstructure was observed by transmission electron microscopy. At the early stage (6-12 hours after hypoxic ischemia) of hypoxic ischemic injury, expression of glutamate transporter excitatory amino acid transporter-2 and synaptophysin was downregulated, the number of synaptic vesicles was reduced, and synaptic swelling was observed; at 12-24 hours after hypoxic ischemia, the Notch pathway was activated, excitatory amino acid transporter-2 and synaptophysin expression was increased, and the number of synaptic vesicles was slightly increased. Excitatory amino acid transporter-2 and synaptophysin expression decreased after treatment with the Notch pathway inhibitor. This suggests that glutamate transport in astrocytes-neurons after hypoxic ischemic injury is regulated by the Notch pathway and affects vesicle release and synaptic plasticity through the expression of synaptophysin.