Neurochemistry international最新文献_第4页

LC-MS/MS techniques for the analysis of steroid panel in human cerebrospinal fluid LC-MS/MS技术分析人脑脊液中类固醇成分。

IF 4 3区医学 Q2 BIOCHEMISTRY & MOLECULAR BIOLOGY

Neurochemistry international

Pub Date : 2025-10-23 DOI: 10.1016/j.neuint.2025.106080

Tereza Skodova , Jana Vitku , Ondrej Bradac , Petr Skalicky , Adela Bubenikova , Radmila Kanceva , Lucie Kolatorova

The metabolic processes within the brain are reflected in the cerebrospinal fluid (CSF). It is in close contact with the nervous system, which is both target and source of multiple steroids. The aim of our study was to develop and validate robust, sensitive LC-MS/MS methods with and without derivatization step for the analysis of unconjugated steroids from all major steroid classes in CSF. The validation of the method without derivatization was performed for ten C19- steroids (dehydroepiandrosterone (DHEA), 7α-hydroxyDHEA, 7β-hydroxyDHEA, 7-ketoDHEA, testosterone, epitestosterone, dihydrotestosterone, 11-hydroxytestosterone, 11-ketotestosterone and androstenedione), ten C21- steroids (cortisol, 11-deoxycortisol, 21-deoxycortisol, cortisone, corticosterone, 11-deoxycorticosterone, pregnenolone, progesterone, 17-hydroxyprogesterone, aldosterone) and three C18- steroids (estrone, estradiol, estriol). The method with derivatization is validated for determination of eleven C19- steroids (testosterone, 11-ketodihydrotestosterone, 11-hydroxytestosterone, DHEA, 7α-hydroxyDHEA, 7β-hydroxyDHEA, 7-ketoDHEA, androstenedione, androsterone, epiandrosterone, 7β-hydroxyepiandrosterone) and six C21- steroids (cortisol, cortisone, corticosterone, pregnenolone, 17-hydroxypregnenolone, progesterone) in CSF. The method without derivatization is applicable for the determination of the majority of steroids in CSF, except for pregnenolone, 17-hydroxypregnenolone and DHEA, for which the derivatization method provides better sensitivity. When analyzing CSF samples of normal pressure hydrocephalus (NPH) patients, 11-ketodihydrotestosterone, epitestosterone, androsterone, epiandrosterone, 7β-hydroxyepiandrosterone, 7-ketoDHEA and 21-deoxycortisol were found to be below the LLOQ, suggesting that their presence is very limited. 17-hydroxypregnenolone, and 11-deoxycortisol were quantified for the first time, their CSF levels in NPH subjects are presented. We also observed significantly increased CSF levels of testosterone and 17-hydroxyprogesterone in men compared to women, both with NPH.

脑内的代谢过程反映在脑脊液（CSF）中。它与神经系统密切接触，是多种类固醇的靶点和来源。本研究的目的是开发和验证可靠、敏感的LC-MS/MS方法，用于分析脑脊液中所有主要类固醇类的非共轭类固醇。对10种C19-甾体（脱氢表雄酮（DHEA）、7α-羟基DHEA、7-羟基DHEA、7-酮DHEA、睾酮、表雄酮、二氢睾酮、11-羟基睾酮、11-酮睾酮和雄烯二酮）、10种C21-甾体（皮质醇、11-脱氧皮质醇、21-脱氧皮质醇、可的松、皮质酮、11-脱氧皮质酮、孕烯醇酮、孕酮、17-羟孕酮、醛固酮）和3种C18-甾体(雌酮、雌二醇、雌三醇)。验证了衍生化法测定脑脊液中11种C19-甾体（睾酮、11-酮二氢睾酮、11-羟基睾酮、DHEA、7α-羟基DHEA、7β-羟基DHEA、雄烯二酮、雄酮、表雄酮、7β-羟基雄酮）和6种C21-甾体（皮质醇、可的松、皮质酮、孕烯醇酮、17-羟基孕烯醇酮、孕酮）的含量。除孕烯醇酮、17-羟基孕烯醇酮和脱氢表雄酮外，不衍生化法适用于脑脊液中大部分类固醇的测定，衍生化法对这些类固醇的灵敏度更高。在分析常压脑积水（NPH）患者脑脊液样本时，发现11-酮二氢睾酮、表甾酮、雄酮、表雄酮、7- β-羟基雄酮、7-酮脱氢表雄酮和21-脱氧皮质醇低于LLOQ，表明它们的存在非常有限。首次定量测定了17-羟基孕烯醇酮和11-脱氧皮质醇，并给出了它们在NPH受试者脑脊液中的水平。我们还观察到，与NPH患者相比，男性脑脊液中睾酮和17-羟孕酮水平显著升高。

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引用次数: 0

Astrocytic HIF-1α/VEGF induces endothelial PI3K/Akt activation to accelerate post-ischemic angiogenesis upon LCN2 inhibition 星形胶质细胞HIF-1α/VEGF诱导内皮细胞PI3K/Akt激活，在LCN2抑制下加速缺血后血管生成。

IF 4 3区医学 Q2 BIOCHEMISTRY & MOLECULAR BIOLOGY

Neurochemistry international

Pub Date : 2025-10-14 DOI: 10.1016/j.neuint.2025.106078

Ning Tian , Xiaoxia Li , Yanlin Jiang , Jungang Deng , Hao Wang , Bing Guo , Meiling Chen , Rujia Liao

Therapeutic angiogenesis represents a pivotal yet underexplored avenue for functional recovery following cerebral ischemia. Although lipocalin-2 (LCN2) participates in neuropathological processes, its cell-type-specific regulation of post-ischemic vascular remodeling remains unknown. Here, we demonstrate that CRISPR/Cas9-mediated C8D1A astrocyte-like cells-specific LCN2 knockout significantly enhances vascular network formation in endothelial co-cultures under oxygen-glucose deprivation/reperfusion (OGD/R). Clinically, elevated LCN2 (GDS4521 dataset) correlates with poor stroke prognosis. Functional analyses revealed that AAV-shRNA-mediated LCN2 knockdown in photothrombotic stroke mice reduced infarct volume, attenuated peri-infarct neuronal loss, increased peri-infarct vascular density, and improved neurobehavioral outcomes at 7 days post-ischemia. Mechanistically, transcriptomic profiling identified hypoxia-inducible factor 1α (HIF-1α) as the master regulator of ischemia-induced angiogenesis. Molecular docking confirmed LCN2-HIF1α interaction. Furthermore, LCN2 ablation unleashes a HIF-1α/VEGF signaling cascade in C8D1A astrocyte-like cells, which activates endothelial phosphatidylinositol-3-kinase/protein kinase B (PI3K/Akt) via paracrine mechanisms to drive functional revascularization. These findings not only redefine ischemic pathophysiology but also pioneers LCN2 inhibition as a translational strategy to overcome the limitations of current pro-angiogenic therapies in cerebrovascular disease.

治疗性血管生成是脑缺血后功能恢复的关键但尚未充分探索的途径。尽管脂载素-2 （LCN2）参与神经病理过程，但其对缺血后血管重构的细胞类型特异性调控尚不清楚。在这里，我们证明了CRISPR/ cas9介导的C8D1A星形细胞样细胞特异性LCN2敲除显著增强了氧糖剥夺/再灌注（OGD/R）下内皮共培养血管网络的形成。临床上，升高的LCN2 （GDS4521数据集）与卒中预后不良相关。功能分析显示，在光血栓性卒中小鼠中，aav - shrna介导的LCN2敲低减少了梗死体积，减轻了梗死周围神经元的损失，增加了梗死周围血管密度，并在缺血后7天改善了神经行为结果。在机制上，转录组学分析发现缺氧诱导因子1α (HIF-1α)是缺血诱导血管生成的主要调节因子。分子对接证实LCN2-HIF1α相互作用。此外，LCN2消融在C8D1A星形细胞样细胞中释放HIF-1α/VEGF信号级联，通过旁分泌机制激活内皮磷脂酰肌醇-3激酶/蛋白激酶B (PI3K/Akt)，驱动功能性血运重建。这些发现不仅重新定义了缺血病理生理学，而且开创了LCN2抑制作为一种翻译策略来克服当前脑血管疾病促血管生成治疗的局限性。

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引用次数: 0

Psychotropic and neurodegenerative drugs modulate platelet activity via the PAF pathway 精神药物和神经退行性药物通过PAF途径调节血小板活性。

IF 4 3区医学 Q2 BIOCHEMISTRY & MOLECULAR BIOLOGY

Neurochemistry international

Pub Date : 2025-10-13 DOI: 10.1016/j.neuint.2025.106073

Savvato Kosidou , Zisis Zannas , Anna Ofrydopoulou , Dimitra A. Lambropoulou , Alexandros Tsoupras

Mild psychiatric conditions such as anxiety and depression, as well as severe disorders like schizophrenia and neurodegenerative diseases, are increasingly recognized as systemic inflammatory conditions. Platelets possess both hemostatic and immunomodulatory roles in these situations, with sharing key molecular pathways with the central nervous system, offering thus a valuable peripheral model for evaluating psychotropic drug effects. Platelet-activating factor (PAF), a potent thrombo-inflammatory mediator, has emerged as a potential link between the two systems, yet its involvement in drug responses remains understudied. This study systematically investigates the effects of psychotropic drugs (i.e. antidepressants, antipsychotics and anxiolytics), and neuroprotective (anti-Alzheimer's/Anti-Parkinson's) drugs on platelet aggregation, focusing on PAF-pathway in comparison to a control platelet agonist, ADP. Using ex vivo light transmission aggregometry, we determined IC₅₀ values for each drug and analyzed the impact of selected drug combinations, in which the NSAID diclofenac was also included. Results revealed that most of the compounds assessed inhibited more effectively the PAF-induced aggregation of platelets compared to their effect on the ADP-pathway, with perphenazine showing the greatest anti-PAF potency. Several drug combinations, notably those including alprazolam and diclofenac, demonstrated significant synergistic effects. These findings suggest that commonly prescribed psychotropic drugs and medications for neurodegenerative disorders can influence platelet activity, mostly through the PAF-pathway, and that their interactions with NSAIDs may amplify their efficacy. Nevertheless, some drugs and their combinations induced lysis of platelets at much higher concentrations than their IC50 values, which stems safety concerns for their use.

焦虑和抑郁等轻度精神疾病，以及精神分裂症和神经退行性疾病等严重疾病，越来越多地被认为是全身性炎症。在这些情况下，血小板具有止血和免疫调节作用，与中枢神经系统共享关键的分子通路，因此为评估精神药物的作用提供了有价值的外周模型。血小板活化因子（PAF）是一种有效的血栓炎症介质，已成为这两个系统之间的潜在联系，但其在药物反应中的作用仍未得到充分研究。本研究系统地研究了精神药物（如抗抑郁药、抗精神病药和抗焦虑药）和神经保护药物（抗阿尔茨海默病/抗帕金森病）对血小板聚集的影响，重点研究了paf途径与对照血小板激动剂ADP的比较。使用体外光透射聚合法，我们确定了每种药物的IC50值，并分析了所选药物组合的影响，其中也包括非甾体抗炎药双氯芬酸。结果显示，与对adp通路的影响相比，大多数被评估的化合物更有效地抑制paf诱导的血小板聚集，其中奋乃嗪显示出最大的抗paf效力。几种药物组合，特别是阿普唑仑和双氯芬酸，显示出显著的协同效应。这些发现表明，常用的精神药物和神经退行性疾病药物可以影响血小板活性，主要是通过paf途径，并且它们与非甾体抗炎药的相互作用可能会增强其疗效。然而，一些药物及其组合在比IC50值高得多的浓度下诱导血小板溶解，这引起了对其使用安全性的担忧。

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引用次数: 0

Neuronal FSTL4 negatively regulates BDNF-mediated neuron-glioma interaction 神经元FSTL4负调控bdnf介导的神经元-胶质瘤相互作用。

IF 4 3区医学 Q2 BIOCHEMISTRY & MOLECULAR BIOLOGY

Neurochemistry international

Pub Date : 2025-10-11 DOI: 10.1016/j.neuint.2025.106072

Yan Sun , Mi Xiao , Xunhui Wang , Yanchun Xu , Anbin Chen , Bin Li , Baohui Feng , Bangbao Tao

Gliomas exploit various molecular pathways to promote their survival, proliferation, and invasion. Recent studies reveal the complex neuron-glioma interaction and BDNF plays a major role in this interaction. However, it's unclear whether and how the BDNF-mediated cross-talk between neurons and gliomas is regulated. FSTL4 is reported to negatively regulate BDNF maturation. Here, we hypothesized that neuronal FSTL4 may negatively regulate BDNF-mediated neuron-glioma cross-talk. By using a combination of approaches like chemogenetic activation of primary neurons and CRISPR knockout/activation of endogenous FSTL4, we show that activated primary neurons support the proliferation of co-cultured glioma cells and neuronal BDNF secretion mediates this neuron-glioma interaction via activating TrkB in glioma cells. In addition, this process is negatively regulated by neuronal FSTL4 as its CRISPR KO in primary neurons further supports the proliferation of co-cultured glioma cells. Importantly, CRISPR activation of endogenous FSTL4 expression in primary neurons results in impaired ability to support co-cultured glioma cells, highlighting the therapeutic potential of activating endogenous FSTL4 for glioma treatment. Taken together, our study shows that the FSTL4/BDNF/TrkB axis plays an essential role in fine-tuning the neuron-glioma interaction and targeting this interplay with CRISPR tools may help to develop novel therapeutic strategies.

胶质瘤利用多种分子途径促进其生存、增殖和侵袭。近年来的研究揭示了复杂的神经-胶质瘤相互作用，而BDNF在这种相互作用中起着重要作用。然而，神经元和胶质瘤之间的bdnf介导的串扰是否以及如何被调节尚不清楚。据报道，FSTL4负调控BDNF成熟。在这里，我们假设神经元FSTL4可能负调控bdnf介导的神经元-胶质瘤串扰。通过化学发生激活原代神经元和CRISPR敲除/激活内源性FSTL4等方法，我们发现激活的原代神经元支持共培养胶质瘤细胞的增殖，神经元BDNF分泌通过激活胶质瘤细胞中的TrkB介导这种神经元与胶质瘤的相互作用。此外，这一过程受到神经元FSTL4的负调控，因为其在原代神经元中的CRISPR KO进一步支持共培养胶质瘤细胞的增殖。重要的是，CRISPR激活原代神经元中内源性FSTL4表达会导致支持共培养胶质瘤细胞的能力受损，这突出了激活内源性FSTL4治疗胶质瘤的治疗潜力。综上所述，我们的研究表明FSTL4/BDNF/TrkB轴在微调神经元-胶质瘤相互作用中起着至关重要的作用，用CRISPR工具靶向这种相互作用可能有助于开发新的治疗策略。

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引用次数: 0

Regional and cell type-specific activation of the unfolded protein response after kainate injection in mice 海碱盐注射后未折叠蛋白反应的区域和细胞特异性激活。

IF 4 3区医学 Q2 BIOCHEMISTRY & MOLECULAR BIOLOGY

Neurochemistry international

Pub Date : 2025-10-09 DOI: 10.1016/j.neuint.2025.106071

Ly Huong Nguyen , Loc Dinh Nguyen , Dat Xuan Dao , Tsuyoshi Hattori , Hiroshi Ishii , Mika Takarada-Iemata , Osamu Hori

The unfolded protein response (UPR) is activated under different neuropathological conditions, such as brain ischemia, epilepsy, and neurodegeneration. We previously reported that a UPR transducer, activating transcription factor 6 (ATF6), and its downstream molecular chaperones in the endoplasmic reticulum (ER) have neuroprotective properties against excitotoxicity. In this study, we examined the temporal and spatial changes in the UPR activation after administration of an excitotoxic reagent, kainate (KA), into mice. RT-qPCR revealed enhanced expression of UPR genes, with peaks either on day 1 or day 3 after intrahippocampal KA injection. The status of the UPR was analyzed using ER stress-activated indicator (ERAI)-transgenic mice, in which the spliced form of XBP-1, downstream of the IRE1 branch of the UPR, can be monitored. ERAI-derived GFP signals were strongly observed in CA3 neurons and moderately observed in dentate gyrus neurons, but not in CA1 neurons, after KA injection. A small portion of the activated astrocytes was also positive for ERAI signals. Further studies revealed that ERAI signals were observed in both the soma and dendrites of neurons in regions with enhanced neuronal activity and resistance to KA toxicity. These results suggest that the UPR may be associated with the neuronal activity and survival after KA injection.

未折叠蛋白反应（UPR）在不同的神经病理条件下被激活，如脑缺血、癫痫和神经变性。我们之前报道了UPR传感器，激活转录因子6 （ATF6）及其下游内质网（ER）中的分子伴侣具有抗兴奋性毒性的神经保护特性。在这项研究中，我们检测了给药兴奋毒性试剂kainate （KA）后小鼠UPR激活的时间和空间变化。RT-qPCR显示UPR基因表达增强，在海马内注射KA后第1天或第3天达到峰值。利用ER应激激活指示剂（ERAI）转基因小鼠分析了UPR的状态，其中可以监测UPR IRE1分支下游的XBP-1的剪接形式。KA注射后，在CA3神经元中观察到erai衍生的GFP信号强烈，在齿状回神经元中观察到中度，但在CA1神经元中没有。小部分活化的星形胶质细胞ERAI信号也呈阳性。进一步的研究表明，ERAI信号在神经元活性增强和KA毒性抵抗区域的神经元体细胞和树突中都观察到。这些结果提示，KA注射后的UPR可能与神经元活动和存活有关。

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引用次数: 0

Crossing the Borders: the amino acid transporter LAT1 (SLC7A5) in the Blood-Brain Barrier 跨越边界：血脑屏障中的氨基酸转运体LAT1 （SLC7A5）。

IF 4 3区医学 Q2 BIOCHEMISTRY & MOLECULAR BIOLOGY

Neurochemistry international

Pub Date : 2025-10-08 DOI: 10.1016/j.neuint.2025.106070

Mariafrancesca Scalise , Raffaella Scanga , Lara Console , Michele Galluccio , Flaviana Marzano , Andrea Magrì , Lorena Pochini , Cesare Indiveri

The blood-brain barrier is an anatomical structure responsible for controlling the flux of nutrients, metabolites, and xenobiotics into and out of the brain. This fundamental function is carried out through the coordinated action of specific ion channels and membrane transporters belonging to the SLC and ABC superfamilies. Indeed, membrane transporter expression in the BBB is less redundant than in other parts of the body. Therefore, any alteration to one of these proteins may pose a threat to the brain. The fifth member of the SLC7 family, which is expressed at the BBB has been the subject of much research over the years. SLC7A5, also known as LAT1, is a plasma membrane transporter of essential amino acids, whose role in brain development is well recognised. The protein is expressed in the membranes of BBB vessels, neurons, and microglia, creating a connection between different areas of the human brain. LAT1 received significant attention in the context of brain tumor treatment, particularly for glioblastoma multiforme, a malignancy with a poor prognosis characterised by fatal relapses. Since several drugs are also substrates of LAT1, its expression at the BBB could be exploited to deliver drugs that target brain diseases. This review describes the functional, structural, and regulatory features of LAT1, focusing on pharmacology in the context of brain homeostasis.

血脑屏障是一种解剖学结构，负责控制营养物质、代谢物和异种生物进出大脑的流动。这一基本功能是通过属于SLC和ABC超家族的特定离子通道和膜转运体的协调作用来实现的。的确，血脑屏障中的膜转运蛋白表达比身体其他部位的表达少得多。因此，这些蛋白质的任何改变都可能对大脑构成威胁。SLC7家族的第五个成员，在BBB表达，多年来一直是许多研究的主题。SLC7A5，也被称为LAT1，是一种必需氨基酸的质膜转运蛋白，其在大脑发育中的作用已得到充分认识。这种蛋白质在血脑屏障、神经元和小胶质细胞的膜上表达，在人类大脑的不同区域之间建立联系。LAT1在脑肿瘤治疗中受到了极大的关注，特别是对于多形性胶质母细胞瘤，这是一种以致命复发为特征的预后不良的恶性肿瘤。由于几种药物也是LAT1的底物，因此可以利用其在血脑屏障上的表达来递送针对脑部疾病的药物。本文介绍了LAT1的功能、结构和调控特征，重点介绍了LAT1在脑内稳态中的药理学作用。

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引用次数: 0

Reprogramming activated astrocytes into GABAergic neurons to treat trigeminal neuralgia 活化星形胶质细胞重编程为gaba能神经元治疗三叉神经痛。

IF 4 3区医学 Q2 BIOCHEMISTRY & MOLECULAR BIOLOGY

Neurochemistry international

Pub Date : 2025-10-08 DOI: 10.1016/j.neuint.2025.106067

Shuangyin Xia , Kai Chen , Xin Li , Dingquan Zou , Meng Wang , Yaping Wang

Trigeminal neuralgia (TN) is the most common type of cranial neuralgia. Currently, there remains a significant gap in the availability of effective and safe treatment options in clinical practice. Transdifferentiation of proliferating activated astrocytes into inhibitory neurons is a potential therapeutic strategy for central nervous system diseases. GABAergic neurons are one of the most type of prevalent inhibitory neurons. This study aims to reprogram proliferating astrocytes in the spinal trigeminal subnucleus caudalis (SpVc) into GABAergic neurons, could improve neuronal excitation-inhibition balance, alleviate pain, which serve as a potential treatment for trigeminal neuralgia. A chronic constriction injury of the distal infraorbital nerve (CCI-dION) was induced in the infraorbital branch of the trigeminal nerve to create a rat model of TN. Adeno-associated viruses were used to overexpress transcription factors Sox2 and Mash1 in astrocytes. The changes in astrocytes and GABAergic neurons in the SpVc region were detected by immunofluorescence, Western blotting, qPCR, and electron microscopy. The mechanical pain threshold testing was used to assess rat TN. In the SpVc region of CCI-dION rats, astrocytes showed proliferation and activation, and the number of GABAergic neurons decreased significantly. Overexpressing Sox2 and Mash1 in astrocytes led to a significant transdifferentiation into GABAergic neurons, which − improved the mechanical pain threshold in CCI-dION rats. Furthermore, fluorocitrate-mediated astrocyte deactivation abolished both the neuronal reprogramming and the analgesic effects, underscoring the essential role of astrocytes in this process. These findings suggest that overexpressing Sox2 and Mash1 in astrocytes led to a significant transdifferentiation into GABAergic neurons, which significantly improved the mechanical pain threshold in CCI-dION rats. Thus, this approach has the potential to provide a new treatment for TN.

三叉神经痛（TN）是最常见的脑神经痛类型。目前，在临床实践中有效和安全的治疗方案的可得性方面仍然存在重大差距。将增殖激活的星形胶质细胞转分化为抑制性神经元是中枢神经系统疾病的一种潜在治疗策略。gaba能神经元是最常见的抑制性神经元之一。本研究旨在将三叉神经痛脊髓尾侧亚核（SpVc）中增殖的星形胶质细胞重编程为gaba能神经元，改善神经元兴奋-抑制平衡，减轻疼痛，为三叉神经痛的治疗提供一种潜在的方法。采用三叉神经眶下支慢性收缩性眶下远端神经（CCI-dION）损伤模型，利用腺相关病毒在星形胶质细胞中过表达转录因子Sox2和Mash1。采用免疫荧光、western blotting、qPCR和电镜观察SpVc区星形胶质细胞和gaba能神经元的变化。采用机械痛阈测试评估大鼠TN。CCI-dION大鼠SpVc区星形胶质细胞增殖活化，gaba能神经元数量明显减少。在星形胶质细胞中过表达Sox2和Mash1，可显著转分化为gaba能神经元，显著提高CCI-dION大鼠的机械痛阈值。此外，氟柠檬酸介导的星形胶质细胞失活消除了神经元重编程和镇痛作用，强调了星形胶质细胞在这一过程中的重要作用。上述结果提示，星形胶质细胞过表达Sox2和Mash1可显著转分化为gaba能神经元，显著提高CCI-dION大鼠的机械痛阈值。因此，这种方法有可能为TN提供一种新的治疗方法。

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引用次数: 0

Histone lactylation is associated with METTL3-dependent LCN2 m6A modification and astrocyte activation after intracerebral hemorrhage 脑出血后，组蛋白乳酸化与mettl3依赖性LCN2 m6A修饰和星形胶质细胞活化有关。

IF 4 3区医学 Q2 BIOCHEMISTRY & MOLECULAR BIOLOGY

Neurochemistry international

Pub Date : 2025-10-06 DOI: 10.1016/j.neuint.2025.106069

Ling Gao , Xiaobin Zheng , Cong Tan , Li Peng , Chuang Wang , Zhongtao Zheng , Jiangli Han , Jian Wang , Zhao Yang , Weiming Chen

Background

Intracerebral hemorrhage (ICH) is a major cause of secondary brain injury (SBI), which results in severe neurological deficits and poor clinical outcomes. Elevated serum lactate levels have been associated with unfavorable outcome in ICH patients. However, the role of lactate in ICH-induced SBI remain poorly understood.

Method

An autologous blood injection mouse model of ICH and lactate-treated C8D1A cells were employed as the in vivo and in vitro models, respectively. The establishment of ICH model was validated by behavior tests, and brain injury was assessed by H&E and Nissel staining. qRT-PCR, Western blot and IHC analysis were used to detect the expression of key molecules. Immunofluorescent (IF) staining was employed to evaluate astrocyte activation. Pro-inflammatory cytokine release was monitored by ELISA assay. The interaction between H3K18la and METTL3 was assessed by ChIP assay, and the association between METTL3 and LCN2 mRNA was assessed by RNA immunoprecipitation (RIP) assay.

Results

The levels of lactate, METTL3 and LCN2 are elevated in ICH model in mice. The inhibition of lactate decreased METTL3 expression and alleviated ICH-induced SBI. Mechanistically, histone H3K18 lactylation was associated with the upregulated levels of METTL3 and m⁶A in mouse brains. METTL3 regulated the m⁶A modification of LCN2 and upregulated its expression. In ICH mice, silencing of LCN2 inhibited A1 astrocyte activation. Histone lactylation-modulated LCN2 m⁶A modification is involved in astrocyte activation and the regulation of SBI in ICH mice.

Conclusion

These results suggested a mechanism whereby histone lactylation is implicated in the activation of A1 astrocytes through METTL3-mediated LCN2 m⁶A modification.

背景：脑出血（Intracerebral hemorrhage， ICH）是继发性脑损伤（secondary brain injury， SBI）的主要原因，可导致严重的神经功能缺损和较差的临床预后。血清乳酸水平升高与脑出血患者的不良预后有关。然而，乳酸盐在ich诱导的SBI中的作用仍然知之甚少。方法：采用自体血液注射小鼠脑出血模型和乳酸处理的C8D1A细胞作为体内模型和体外模型。行为学实验验证脑缺血模型的建立，H&E和Nissel染色评价脑损伤程度。采用qRT-PCR、western blot和免疫组化分析检测关键分子的表达。免疫荧光（IF）染色评价星形胶质细胞活化。ELISA法检测促炎细胞因子释放情况。采用ChIP法评估H3K18la与METTL3的相互作用，采用RNA免疫沉淀（RIP）法评估METTL3与LCN2 mRNA的相关性。结果：小鼠脑出血模型中乳酸、METTL3、LCN2水平升高。乳酸的抑制降低了METTL3的表达，减轻了ich诱导的SBI。机制上，组蛋白H3K18乳酸化与小鼠大脑中METTL3和m6A水平上调有关。METTL3调控LCN2的m6A修饰，上调其表达。在脑出血小鼠中，LCN2的沉默抑制了A1星形胶质细胞的激活。组蛋白乳酸化调控的LCN2 m6A修饰参与脑出血小鼠星形细胞活化和SBI的调控。结论：这些结果提示了一种机制，即组蛋白乳酸化通过mettl3介导的LCN2 m6A修饰参与A1星形胶质细胞的激活。

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引用次数: 0

Differences in presynaptic hippocampal GABAergic terminals at the early stage of life in female and male mice: effect of an acute early inflammatory challenge 雌性和雄性小鼠早期海马突触前gaba能终端的差异：急性早期炎症的影响

IF 4 3区医学 Q2 BIOCHEMISTRY & MOLECULAR BIOLOGY

Neurochemistry international

Pub Date : 2025-10-04 DOI: 10.1016/j.neuint.2025.106062

Cristina Benatti , Alessandra Roggeri , Ylenia Toscano , Veronica Torre , Nicoletta Brunello , Fabio Tascedda , Johanna Maria Catharina Blom , Anna Pittaluga

GABA dictates the efficiency of synaptic connection, influencing its developmental complexity, but its role is tuned by developmental sex differences which affect the efficiency of its innervation. We investigated the efficiency of mechanisms of GABA storage and exocytosis in hippocampal terminals of male and female mice during the juvenile period (PND21), adolescence (PND36) or adulthood (PND90). The expression of mRNA encoding for the presynaptic GABA transporter type 1, (GAT1) and the vesicular GABA transporter (VGAT1) was analysed. A significant scaling-down in the GAT1 mRNA levels (SLC6A1) was detected at PND21 in both sexes until adulthood, while the SLC32A1-VGAT mRNA level was conserved. We also analysed the density of GAT1 and VGAT proteins. Western blot analysis unveiled the presence of a monomeric and an oligomeric form of GAT1. The density of the monomeric form was conserved at the different stages of development in both sexes. Differently, the oligomeric assembly was significantly overexpressed in hippocampal synaptosomal lysates from PND21 male and female mice, but recovered at PND36. VGAT density was largely conserved in PND21 and PND36 male hippocampal synaptosomal lysates when compared to adult particles, but significantly lower in PND21 female particles. Notably, these changes are consistent and support the altered vesicular storage of newly taken-up [³H]GABA detected in PND21 male and female hippocampal synaptosomes as well as the different responsiveness of GABAergic male and female synaptosomes to increasing depolarizing stimuli (12, 20 and 30 mM KCl-enriched solutions) measured as efficiency of the [³H]GABA exocytosis. Interstingly, an acute LPS treatment affects the efficiency of GABA exocytosis at PND36 in a sex-dependent manner. These results add new knowledge on the role of GABA as effector of central inhibitory plasticity at the early stage of development and its relevance in dimorphic adaptation in physio pathological conditions.

GABA决定突触连接的效率，影响其发育的复杂性，但其作用受发育性别差异的调节，而发育性别差异影响其神经支配的效率。我们研究了雄性和雌性小鼠在幼年期（PND21）、青春期（PND36）和成年期（PND90）海马末端GABA储存和胞外分泌的效率机制。分析突触前GABA转运蛋白1型（GAT1）和囊状GABA转运蛋白VGAT1的mRNA表达。在PND21，直到成年，两性都检测到GAT1 mRNA水平（SLC6A1）的显著下降，而SLC32A1-VGAT mRNA水平则保持不变。我们还分析了GAT1和VGAT蛋白的密度。Western blot分析揭示了GAT1的单体和寡聚形式的存在。在两性的不同发育阶段，单体形态的密度是保守的。不同的是，在PND21雄性和雌性小鼠的海马突触体溶解物中，低聚物组装显著过表达，但在PND36时恢复。与成体颗粒相比，PND21和PND36雄性海马突触体溶解物中的VGAT密度基本保守，但PND21雌性颗粒中的VGAT密度明显降低。值得注意的是，这些变化是一致的，并支持PND21雄性和雌性海马突触体中新摄取的[3H]GABA的囊泡储存改变，以及GABA能雄性和雌性突触体对增加的去极化刺激（12、20和30 mM氯化钾富集溶液）的不同反应性（以[3H]GABA胞分泌效率衡量）。有趣的是，急性LPS治疗以性别依赖的方式影响PND36的GABA胞吐效率。这些结果为GABA在发育早期中枢抑制性可塑性的作用及其在生理病理条件下二形适应的相关性提供了新的认识。

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引用次数: 0

TREM2 deficiency aggravates neuroinflammatory response and cognitive impairment via disease-associated microglia in Parkinson's disease models 在帕金森病模型中，TREM2缺乏通过疾病相关的小胶质细胞加重神经炎症反应和认知障碍。

IF 4 3区医学 Q2 BIOCHEMISTRY & MOLECULAR BIOLOGY

Neurochemistry international

Pub Date : 2025-10-04 DOI: 10.1016/j.neuint.2025.106068

Zhang Piao , Zhu Baoyu , Feng Jiezhu , Liang Xiaomei , Huang Peiting , He Chentao , Deng Yiyu , Lu Jiahong , Wang Lijuan , Zhang Yuhu

This study explores whether Triggering Receptor Expressed on Myeloid Cells 2 (TREM2) regulates the distinct disease-related microglia (DAM) phenotype and exerts a protective role in cognitive impairment in Parkinson's disease (PD). Adeno-associated virus carrying TREM2 shRNA (AAV-TREM2-shRNA) was injected into the bilateral hippocampus of the A53T α-Synuclein (α-Syn) transgenic PD mouse model; Additionally, lentivirus was transduced into BV2 microglial cells to knock out the expression of TREM2, which were subsequently stimulated with α-Syn preformed fibrils (PFF). Furthermore, cognitive status of mice, α-Syn aggregation, microglia status, expression of inflammatory factors, pro-inflammatory and anti-inflammatory DAM markers, MAPK and NF- κB pathway activation status and neuron apoptosis were evaluated. TREM2 deficiency induced cognitive impairment in A53T α-Syn PD mice by decreased performance in the novel objective recognition and Morris water maze tests. TREM2 knockdown resulted in synaptic loss, microglial activation, increased inflammatory factors, and MAPK and NF- κB pathway activation in the hippocampus of mice. In vitro, TREM2 deficiency exacerbated the inflammatory response of BV2 cells stimulated by α-Syn PFF by inhibiting anti-inflammatory DAM, and promoting neuronal apoptosis and Ser129-phosphorylation of α-Syn. TREM2 knockdown also promoted pro-inflammatory DAM activation and increased inflammatory factors expression via the ERK1/2 signaling pathway. Our findings suggest that TREM2 plays a protective role in cognitive impairment and promotes anti-inflammatory DAM activation via the ERK1/2 signaling pathway in PD mice, providing novel insight into the immunopathogenesis of cognitive impairments in PD.

本研究探讨骨髓细胞触发受体2 （TREM2）是否调节不同的疾病相关小胶质细胞（DAM）表型，并在帕金森病（PD）的认知障碍中发挥保护作用。将携带TREM2 shRNA的腺相关病毒（AAV-TREM2-shRNA）注射到A53T α-突触核蛋白（α-Syn）转基因PD小鼠模型的双侧海马；此外，将慢病毒转导到BV2小胶质细胞中，敲除TREM2的表达，随后用α-Syn预形成原纤维（PFF）刺激TREM2。进一步观察小鼠的认知状态、α-Syn聚集、小胶质细胞状态、炎症因子表达、促炎和抗炎DAM标志物、MAPK和NF- κB通路激活状态和神经元凋亡。TREM2缺乏导致A53T α-Syn PD小鼠在新型客观识别和Morris水迷宫测试中的表现下降，从而导致认知障碍。TREM2敲低导致小鼠海马突触丢失、小胶质细胞激活、炎症因子增加、MAPK和NF- κB通路激活。在体外，TREM2缺乏通过抑制抗炎DAM，促进神经元凋亡和α-Syn ser129磷酸化，加重α-Syn PFF刺激的BV2细胞的炎症反应。TREM2敲低还通过ERK1/2信号通路促进促炎DAM激活和炎症因子表达增加。我们的研究结果表明，TREM2在PD小鼠认知障碍中具有保护作用，并通过ERK1/2信号通路促进抗炎DAM的激活，为PD认知障碍的免疫发病机制提供了新的见解。

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引用次数: 0