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PX-478 induces apoptosis in acute myeloid leukemia under hypoxia by inhibiting the PI3K/AKT/mTOR pathway through downregulation of GBE1. PX-478 通过下调 GBE1 抑制 PI3K/AKT/mTOR 通路,从而诱导缺氧条件下急性髓性白血病患者的细胞凋亡。
IF 5.3 2区 医学 Q1 PHARMACOLOGY & PHARMACY Pub Date : 2024-11-09 DOI: 10.1016/j.bcp.2024.116620
Wenjing Liu, Chunhui Dou, Ce Zhang, Ping Chen, Shu Zhang, Renxiang Wang, Qing Han, Hongyu Zhao, Daqi Li

Acute myeloid leukemia (AML) is a highly heterogeneous hematologic malignancy characterized by limited therapeutic options and a pronounced tendency for relapse. PX-478, a novel inhibitor of hypoxia-inducible factor 1-alpha (HIF-1α), has demonstrated antitumor activity across various cancer models, but its specific role in AML remains unexplored. This study aimed to explore the potential target and mechanism of PX-478-induced AML cell apoptosis. First, PX-478 induced AML cell apoptosis in vitro under hypoxia via modulation of the Bcl-2 family and activation of the mitochondria-mediated caspase cascade, exhibiting a concentration-dependent effect. Additionally, in vivo administration of PX-478 led to notable inhibition of subcutaneous AML xenograft growth in mice, coupled with increased tumor cell apoptosis. RNA sequencing and cellular studies revealed downregulation of the PI3K/AKT/mTOR signaling pathway in PX-478-treated cells. Consistently, cellular studies also implicated PI3K/AKT/mTOR pathway in PX-478-induced AML cell apoptosis. Furthermore, by screening for RNA sequencing differential genes and subsequent experimental verification, Glycogen branching enzyme 1 (GBE1) may be involved in PX-478-induced apoptosis in AML cells. We found that inhibiting GBE1 expression in AML cells (siGBE1) led to downregulation of the PI3K/AKT/mTOR pathway and induced apoptosis. In experiments using AML cells with reduced GBE1 expression (shGBE1), PX-478 treatment did not further downregulate the pathway or enhance apoptosis. Re-expression of GBE1 in shGBE1 cells alleviated apoptosis and reduced PX-478- induced apoptosis and pathway downregulation. In conclusion, our findings provide convincing evidence that PX-478 induces apoptosis by inhibiting the PI3K/AKT/mTOR pathway through downregulation of GBE1 in AML cells.

急性髓性白血病(AML)是一种高度异质性的血液系统恶性肿瘤,其特点是治疗方案有限且有明显的复发倾向。PX-478是一种新型低氧诱导因子1-α(HIF-1α)抑制剂,已在多种癌症模型中显示出抗肿瘤活性,但其在急性髓性白血病中的具体作用仍有待探索。本研究旨在探索PX-478诱导AML细胞凋亡的潜在靶点和机制。首先,在体外缺氧条件下,PX-478通过调节Bcl-2家族和激活线粒体介导的caspase cascade诱导AML细胞凋亡,表现出浓度依赖性效应。此外,体内给药 PX-478 能显著抑制小鼠皮下急性髓细胞白血病异种移植的生长,同时增加肿瘤细胞的凋亡。RNA 测序和细胞研究显示,PX-478 处理的细胞中 PI3K/AKT/mTOR 信号通路下调。同样,细胞研究也表明 PI3K/AKT/mTOR 通路与 PX-478 诱导的 AML 细胞凋亡有关。此外,通过对 RNA 测序差异基因的筛选和随后的实验验证,糖原分支酶 1(GBE1)可能参与了 PX-478 诱导的 AML 细胞凋亡。我们发现,抑制 AML 细胞中 GBE1 的表达(siGBE1)会导致 PI3K/AKT/mTOR 通路下调并诱导细胞凋亡。在使用减少了 GBE1 表达(shGBE1)的 AML 细胞进行的实验中,PX-478 处理并未进一步下调通路或增强细胞凋亡。在 shGBE1 细胞中重新表达 GBE1 可缓解细胞凋亡,减少 PX-478 诱导的细胞凋亡和通路下调。总之,我们的研究结果提供了令人信服的证据,证明PX-478通过抑制AML细胞中的GBE1,抑制PI3K/AKT/mTOR通路,从而诱导细胞凋亡。
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引用次数: 0
Evaluation of UCP1162, a potent propargyl-linked inhibitor of dihydrofolate reductase with potential application to cancer and autoimmune disease 对 UCP1162 进行评估,它是一种有效的二氢叶酸还原酶丙炔链抑制剂,有望应用于癌症和自身免疫性疾病。
IF 5.3 2区 医学 Q1 PHARMACOLOGY & PHARMACY Pub Date : 2024-11-09 DOI: 10.1016/j.bcp.2024.116617
Didem Ozcan Tezgin , Shan Kurkcu , Debjani Si , Jolanta Krucinska , Adriane Mosley , Pratik Mehta , Ivan Babic , Elmar Nurmemmedov , Alan Kuo , Wu He , Craig E Nelson , Lee Wright , Dennis L. Wright , Charles Giardina
Cellular resistance can limit the effectiveness of antifolate drugs for the treatment of cancer and autoimmune diseases. We examined the biochemical and cellular effects of a propargyl linked, non-classical antifolate UCP1162 that shows exceptional potency and resilience in the background of methotrexate resistance. UCP1162 inhibited the human DHFR enzyme with affinity and kinetics comparable to methotrexate (MTX). UCP1162 also inhibited cancer cell proliferation and bound cellular DHFR at low nanomolar concentrations. Leucovorin suppressed the cellular effects of UCP1162, consistent with UCP1162 working as an antifolate. Like other antifolates, UCP1162 reduced acute inflammation in mice and inhibited FLS cell growth and motility. Single cell RNA-seq showed that MTX and UCP1162 generated overlapping gene expression changes after a 48-hour exposure. However, while leukemia cells (CCRF-CEM) resistant to MTX could be readily selected, UCP1162-resistant cells could not be obtained. Long-term exposure to UCP1162 resulted in static culture expressing stem cell genes (CD34, ABCG2, ABCB1), adaptive genes (TCN2, CDKN1A), and genes that might serve as therapeutic targets (TPBG/5T4, TNFRSF10A, ACE). These findings suggest that UCP1162 is a unique tool for studying cellular responses to long-term antifolate treatment and holds promise as a lead compound capable of overcoming some forms of antifolate resistance.
细胞抗药性会限制抗叶酸药物治疗癌症和自身免疫性疾病的效果。在这里,我们研究了一种丙炔基连接的非经典抗叶酸药 UCP1162 的生化和细胞效应,这种药在甲氨蝶呤耐药性背景下显示出卓越的效力和复原力。UCP1162 可抑制人类 DHFR 酶,其亲和力和动力学特性与甲氨蝶呤(MTX)相当。UCP1162 还能抑制癌细胞增殖,并在低纳摩尔浓度下与细胞 DHFR 结合。亮菌甲素抑制了 UCP1162 对细胞的作用,这与 UCP1162 作为一种抗叶酸的作用是一致的。与其他抗叶酸盐一样,UCP1162 也能减轻小鼠的急性炎症反应,抑制 FLS 细胞的生长和运动。单细胞 RNA 序列分析表明,MTX 和 UCP1162 在暴露 48 小时后会产生重叠的基因表达变化。然而,虽然可以很容易地筛选出抗 MTX 的白血病细胞(CCRF-CEM),但却无法获得抗 UCP1162 的细胞。长期暴露于 UCP1162 会导致静态培养表达干细胞基因(CD34、ABCG2、ABCB1)、适应性基因(TCN2、CDKN1A)和可能作为治疗靶点的基因(TPBG/5T4、TNFRSF10A、ACE)。这些研究结果表明,UCP1162 是研究细胞对长期抗叶酸治疗反应的一种独特工具,有望成为能够克服某些形式抗叶酸耐药性的先导化合物。
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引用次数: 0
Detailed functional characterization of four nanobodies as positive allosteric modulators of the human Calcium-Sensing receptor. 四种纳米抗体作为人类钙传感受体正异构调节剂的详细功能表征。
IF 5.3 2区 医学 Q1 PHARMACOLOGY & PHARMACY Pub Date : 2024-11-08 DOI: 10.1016/j.bcp.2024.116619
Wei Du, Sabrina N Rahman, Eleanor Barker, Hans Bräuner-Osborne, Jesper M Mathiesen, Donald T Ward, Anders A Jensen

The calcium-sensing receptor (CaSR) plays a key role in calcium homeostasis, and small-molecule and peptide positive allosteric modulators (PAMs) of CaSR, so-called calcimimetics, are used in the treatment of hyperparathyroidism and hypocalcemic disorders. In this study, four monovalent nanobodies - representing four distinct nanobody families with CaSR PAM activity - were subjected to elaborate pharmacological profiling at the receptor. While Nb5 displayed negligible PAM activity at CaSR in all assays, Nb4, Nb10 and Nb45 all potently potentiated Ca2+-evoked signalling through a myc epitope-tagged CaSR expressed in HEK293 or HEK293T cells in Gαq and Gαi1 protein activation assays and in a Ca2+/Fluo-4 assay. Nb4 and Nb10 also displayed comparable PAM properties at a stable CaSR-HEK293 cell line in a Ca2+/Fura-2 imaging assay, but surprisingly Nb45 was completely inactive at this cell line in both the Ca2+/Fura-2 and Ca2+/Fluo-4 assays. Investigations into this binary difference in Nb45 activity revealed that the nanobody only possesses modulatory activity at CaSRs tagged N-terminally with various epitopes (myc, HA, Flag-SNAP), whereas it is inactive at the untagged wild-type receptor. In conclusion, overall each of the nanobodies exhibit similar CaSR PAM properties in a range of assays, and thus none of them display pathway bias as modulators. However, of the four nanobodies Nb4 and Nb10 would be applicable as pharmacological tools for the wild-type CaSR, and the complete inactivity of Nb45 at the untagged CaSR serves as an reminder that epitope-tagging of a receptor, even if deemed functionally silent, can have profound implications for ligand discovery efforts.

钙传感受体(CaSR)在钙平衡中起着关键作用,CaSR 的小分子和多肽正性异位调节剂(PAMs),即所谓的降钙药,被用于治疗甲状旁腺功能亢进症和低钙血症。在这项研究中,对四种单价纳米抗体(代表具有 CaSR PAM 活性的四个不同纳米抗体家族)在受体上进行了详细的药理学分析。在 Gαq 和 Gαi1 蛋白激活试验以及 Ca2+/Fluo-4 试验中,Nb5 在所有试验中对 CaSR 的 PAM 活性都微乎其微,而 Nb4、Nb10 和 Nb45 都能通过在 HEK293 或 HEK293T 细胞中表达的 myc 表位标记的 CaSR 有效地增强 Ca2+ 诱导的信号传导。在 Ca2+/Fura-2 成像检测中,Nb4 和 Nb10 在稳定的 CaSR-HEK293 细胞系中也显示出相似的 PAM 特性,但令人惊讶的是,在 Ca2+/Fura-2 和 Ca2+/Fluo-4 检测中,Nb45 在该细胞系中完全没有活性。对 Nb45 活性的这种二元差异进行研究后发现,纳米抗体只对 N 端标记了各种表位(myc、HA、Flag-SNAP)的 CaSR 具有调节活性,而对未标记的野生型受体则没有活性。总之,总体而言,每种纳米抗体在一系列检测中都表现出相似的 CaSR PAM 特性,因此它们都没有显示出作为调节剂的途径偏差。然而,在这四种纳米抗体中,Nb4 和 Nb10 可以作为野生型 CaSR 的药理工具,而 Nb45 在未标记的 CaSR 上完全没有活性,这提醒我们,即使认为受体的表位标记在功能上是沉默的,也会对配体的发现工作产生深远的影响。
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引用次数: 0
Heat shock protein 27 downregulation attenuates isoprenaline-induced myocardial fibrosis and diastolic dysfunction by modulating the endothelial-mesenchymal transition 通过调节内皮-间充质转化,下调热休克蛋白27可减轻异丙肾上腺素诱导的心肌纤维化和舒张功能障碍。
IF 5.3 2区 医学 Q1 PHARMACOLOGY & PHARMACY Pub Date : 2024-11-07 DOI: 10.1016/j.bcp.2024.116612
Yifei Zou, Henghe Shi, Yinghao Li, Tianyi Li, Ning Liu , Bin Liu
Heart failure (HF), an end-stage clinical syndrome secondary to cardiac impairment, significantly affects patients’ quality of life and long-term prognosis. Myocardial fibrosis leads to systolic and diastolic dysfunction, and promotes the progression of HF. Several studies involving the modulation of myocardial fibrosis have been conducted in an effort to improve cardiac function. Heat shock protein 27 (HSP27) is a small chaperone protein that is overexpressed in cellular stress states. HSP27 modulates epithelial-mesenchymal transition, playing a crucial role in the pathology of several fibrotic diseases. However, its association with myocardial fibrosis regulation is unknown. This study aimed to investigate the mechanisms by which HSP27 regulates myocardial fibrosis. We created cardiac-specific HSP25 (the murine ortholog of human HSP27) knockout mice and found that HSP25 knockdown inhibited endothelial-mesenchymal transition (EndMT), attenuated myocardial fibrosis, and ameliorated diastolic dysfunction in isoproterenol-induced HF mice via echocardiography, histology, and western bloting. In vitro, HSP27 knockdown attenuated transforming growth factor beta-induced EndMT, whereas HSP27 overexpression promoted EndMT. Furthermore, the SMAD3/SNAIL1 pathway was found to be crucial for HSP27-mediated EndMT regulation. As an essential molecule in EndMT regulation and myocardial fibrosis modulation, HSP27 may hold promise as a therapeutic target for patients with HF.
心力衰竭(HF)是继发于心脏功能损害的终末期临床综合征,严重影响患者的生活质量和长期预后。心肌纤维化会导致收缩和舒张功能障碍,并促进心力衰竭的恶化。为了改善心脏功能,已经开展了多项有关调节心肌纤维化的研究。热休克蛋白 27(HSP27)是一种小型伴侣蛋白,在细胞应激状态下会过度表达。HSP27 可调节上皮-间质转化,在多种纤维化疾病的病理过程中发挥关键作用。然而,它与心肌纤维化调控的关系尚不清楚。本研究旨在探讨HSP27调控心肌纤维化的机制。我们创建了心脏特异性 HSP25(人类 HSP27 的鼠直向同源物)基因敲除小鼠,并通过超声心动图、组织学和 Western bloting 发现,HSP25 基因敲除抑制了内皮-间质转化(EndMT),减轻了心肌纤维化,并改善了异丙托肾上腺素诱导的高频小鼠的舒张功能障碍。在体外,HSP27敲除可减轻转化生长因子β诱导的内膜种植,而HSP27过表达则可促进内膜种植。此外,研究还发现SMAD3/SNAIL1通路对HSP27介导的EndMT调控至关重要。作为内膜移植调控和心肌纤维化调节的重要分子,HSP27有望成为心房颤动患者的治疗靶点。
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引用次数: 0
Yohimbine treatment improves pulmonary fibrosis by attenuating the inflammation and oxidative stress via modulating the MAPK pathway 育亨宾通过调节 MAPK 通路减轻炎症和氧化应激,从而改善肺纤维化。
IF 5.3 2区 医学 Q1 PHARMACOLOGY & PHARMACY Pub Date : 2024-11-06 DOI: 10.1016/j.bcp.2024.116613
Vaishnavi Kambhampati , Abhisheik Eedara , Sai Balaji Andugulapati
Idiopathic pulmonary fibrosis (IPF) is a devastating interstitial lung disorder characterized by the accumulation of extracellular matrix and collagen, resulting in significant parenchymal scarring and respiratory failure that leads to mortality. Yohimbine (YBH) is an α-2 adrenergic receptor antagonist with anti-oxidant and anti-inflammatory properties. In the current study, we aimed to investigate the anti-inflammatory, anti-oxidant and anti-fibrotic activity of YBH against LPS/TGF-β-induced differentiation in BEAS-2B/LL29 cells and bleomycin (BLMN) induced pulmonary fibrosis model in rats. Network pharmacology, gene expression, Western-blot analysis, immune-cytochemistry/immunohistochemistry, lung functional analysis, and histology techniques were used to assess the fibrotic marker expression/levels in cells or rat lung tissues. YBH treatment significantly attenuated the LPS-induced pro-inflammatory (identified through a network-pharmacology approach) and oxidative stress markers expression in lung epithelial cells. TGF-β stimulation significantly elevated the fibrotic cascade of markers and treatment with YBH attenuated these markers’ expression/levels. Intra-tracheal administration of BLMN caused a significant elevation of various inflammatory/oxidative stress and fibrotic markers expression in lung tissues and treatment with YBH significantly mitigated the same. Ashcroft score analysis revealed that BLMN exhibited severe distortion of the lungs, elevation of thickness of the alveolar walls and accumulation of collagen in tissues, further treatment with YBH significantly suppressed these events and improved the lung architecture. Lung functional parameters demonstrated that BLMN-induced stiffness and resistance were reduced considerably upon YBH treatment and restored lung function dose-dependently. Overall, this study reveals that YBH treatment significantly attenuated the BLMN-induced fibrosis by regulating the MAPK pathway and provided insightful information for progressing towards translational outcomes.
特发性肺纤维化(IPF)是一种破坏性的间质性肺部疾病,其特点是细胞外基质和胶原蛋白堆积,导致肺实质严重瘢痕形成和呼吸衰竭,从而导致死亡。育亨宾(YBH)是一种α-2肾上腺素能受体拮抗剂,具有抗氧化和抗炎特性。本研究旨在探讨育亨宾对LPS/TGF-β诱导的BEAS-2B/LL29细胞分化和博莱霉素(BLMN)诱导的大鼠肺纤维化模型的抗炎、抗氧化和抗纤维化活性。采用网络药理学、基因表达、Western-blot分析、免疫细胞化学/免疫组织化学、肺功能分析和组织学技术来评估细胞或大鼠肺组织中纤维化标志物的表达/水平。YBH治疗可明显减轻LPS诱导的肺上皮细胞促炎(通过网络药理学方法确定)和氧化应激标记物的表达。TGF-β 刺激可显著提高纤维化级联标记物的表达,而 YBH 治疗可减轻这些标记物的表达/水平。气管内给药 BLMN 会导致肺组织中各种炎症/氧化应激和纤维化标志物的表达明显升高,而使用 YBH 治疗则会明显减轻这种情况。Ashcroft 评分分析表明,BLMN 表现出肺部严重变形、肺泡壁厚度增加和组织中胶原蛋白堆积,进一步使用 YBH 治疗可明显抑制这些现象并改善肺部结构。肺功能参数表明,经 YBH 治疗后,BLMN 引起的肺僵化和阻力大大降低,肺功能的恢复与剂量有关。总之,本研究揭示了 YBH 治疗可通过调节 MAPK 通路显著减轻 BLMN 诱导的纤维化,并为取得转化成果提供了有价值的信息。
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引用次数: 0
Activated microglia secretome and proinflammatory cytokines increase neuronal mu-opioid receptor signalling and expression. 激活的小胶质细胞分泌组和促炎细胞因子会增加神经元μ-阿片受体的信号传导和表达。
IF 5.3 2区 医学 Q1 PHARMACOLOGY & PHARMACY Pub Date : 2024-11-06 DOI: 10.1016/j.bcp.2024.116608
Javier Cuitavi, Pere Duart-Abadia, Julie Sanchez, Christian M Sánchez-López, Jesús D Lorente, Antonio Marcilla, Isabel Fariñas, Meritxell Canals, Lucía Hipólito

Due to its potential role in processes which rely on mu-opioid receptor function, investigating the relationship between Mu-Opioid receptors (MORs), neuroinflammation, and glial cells has gained momentum. Traditionally, MOR activation has been associated with immunosuppression, but recent findings suggest a more nuanced, bidirectional relationship with the immune system. To further investigate this relationship, herein, we investigated the role of the activated microglia secretome and proinflammatory cytokines in neuronal MOR expression and signalling. Our results show that both microglial secretome and specific cytokines increase neuronal MOR expression and enhance the [D-Ala2, N-MePhe4, Gly-ol]-enkephalin (DAMGO)-induced MOR activation. We also show that DAMGO-induced neuroinflammation increases neuronal MOR expression, activation, and regulation. Our findings suggest a feedback loop between microglial activation, cytokine release, and neuronal MOR dynamics. Future research should delve into the temporal dynamics and functional implications of this relationship, particularly concerning clinically relevant opioids like morphine and fentanyl and pain management.

由于μ-阿片受体在依赖μ-阿片受体功能的过程中可能发挥作用,研究μ-阿片受体(MOR)、神经炎症和神经胶质细胞之间关系的势头日益强劲。传统上,MOR 的激活与免疫抑制有关,但最近的研究结果表明,MOR 与免疫系统之间存在着更加微妙的双向关系。为了进一步研究这种关系,我们在本文中研究了活化的小胶质细胞分泌组和促炎细胞因子在神经元 MOR 表达和信号传导中的作用。我们的研究结果表明,小胶质细胞分泌物和特定细胞因子都会增加神经元 MOR 的表达,并增强[D-Ala2, N-MePhe4, Gly-ol]-enkephalin (DAMGO)诱导的 MOR 激活。我们还发现,DAMGO 诱导的神经炎症会增加神经元 MOR 的表达、激活和调节。我们的研究结果表明,小胶质细胞活化、细胞因子释放和神经元 MOR 动态之间存在反馈回路。未来的研究应深入探讨这种关系的时间动态和功能影响,特别是与吗啡和芬太尼等临床相关的阿片类药物和疼痛治疗。
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引用次数: 0
Niclosamide attenuates calcification in human heart valvular interstitial cells through inhibition of the AMPK/mTOR signaling pathway 尼可刹米通过抑制 AMPK/mTOR 信号通路减轻人类心脏瓣膜间质细胞的钙化。
IF 5.3 2区 医学 Q1 PHARMACOLOGY & PHARMACY Pub Date : 2024-11-06 DOI: 10.1016/j.bcp.2024.116614
Minh Hung Vu , Saugat Shiwakoti , Ju-Young Ko , Geul Bang , Eunmi Lee , Eunmin Kim , Sin-Hee Park , Eun-Hye Park , Chan Woo Kim , Jin Young Kim , Hwan-Hee Sim , Kiyuk Chang , Min-Sik Kim , Min-Ho Oak
Calcific aortic valve disease (CAVD) is a considerable health burden with a lack of effective therapeutic options. There is an urgent need to develop interventions that inhibit the osteogenic transformation of valvular interstitial cells (VICs) and delay the calcification process. Niclosamide, an FDA-approved anti-helminthic drug, has emerged as a promising candidate that demonstrates a negative regulatory effect on porcine VICs calcification. However, its molecular mechanism in human VICs (hVICs) remains to be investigated. In this study, high-resolution mass spectrometry-based proteomics and phosphoproteomics were employed, and 8373 proteins and 3697 phosphosites were identified in hVICs treated with a pro-calcifying medium and niclosamide. The quantitative proteomic and phosphoproteomic analysis resulted in the identification of calcification markers and osteogenesis-associated proteins. Bioinformatic analysis of the protein–protein interaction network and affected kinase prediction revealed that the AMPK/mTOR/p70S6K signaling cascade was altered upon calcific induction and niclosamide treatment. Further validation indicated that niclosamide inhibited the calcification of hVICs by targeting the mammalian target of the rapamycin (mTOR) signaling pathway. This study provides the first evidence that niclosamide could prevent osteoblastic differentiation in hVICs partially through the inhibition of the AMPK/mTOR/p70S6k signaling pathway, thereby mitigating hVICs calcification. These findings present a foundation for potential therapeutic strategies to impede the progression of CAVD and provide valuable insights into the pharmacological effects of niclosamide on human VICs.
钙化性主动脉瓣病(CAVD)是一种严重的健康负担,目前缺乏有效的治疗方案。目前迫切需要开发干预措施,抑制瓣膜间质细胞(VICs)的成骨转化,延缓钙化过程。尼可刹米是美国食品及药物管理局批准的一种抗蠕虫药物,它对猪瓣膜间质细胞的钙化具有负面调节作用,是一种很有前景的候选药物。然而,它在人VICs(hVICs)中的分子机制仍有待研究。本研究采用基于高分辨质谱的蛋白质组学和磷酸化蛋白质组学方法,在使用促钙化培养基和烟酰胺处理的人VICs中鉴定出8373个蛋白质和3697个磷酸化位点。定量蛋白质组学和磷酸化蛋白质组学分析鉴定了钙化标志物和成骨相关蛋白。蛋白-蛋白相互作用网络的生物信息学分析和受影响的激酶预测显示,AMPK/mTOR/p70S6K 信号级联在钙化诱导和尼可洛沙胺处理后发生了改变。进一步的验证表明,尼可刹米通过靶向雷帕霉素哺乳动物靶标(mTOR)信号通路抑制了 hVICs 的钙化。这项研究首次证明,尼可刹米可通过抑制AMPK/mTOR/p70S6k信号通路阻止hVICs的成骨细胞分化,从而减轻hVICs的钙化。这些发现为制定潜在的治疗策略以阻止 CAVD 的进展奠定了基础,并为了解烟酰胺对人类 VICs 的药理作用提供了宝贵的见解。
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引用次数: 0
Chemical proteomics accelerates the target discovery of natural products 化学蛋白质组学加速了天然产品的目标发现。
IF 5.3 2区 医学 Q1 PHARMACOLOGY & PHARMACY Pub Date : 2024-11-05 DOI: 10.1016/j.bcp.2024.116609
Shu-Jie He , Jun Li , Jie-Chun Zhou , Zhi-You Yang , Xi Liu , Yue-Wei Ge
More than half of the global novel drugs are directly or indirectly derived from natural products (NPs) because of their better selectivity towards proteins. Traditional medicines perform multiple bioactivities through various NPs binding to drug targets, which highlights the opportunities of target discovery for drug development. However, detecting the binding relationship between NPs and targets remains challenging. Chemical proteomics, an interdisciplinary field of chemistry, proteomics, biology, and bioinformatics, has emerged as a potential approach for uncovering drug-target interactions. This review summarizes the principles and characteristics of the current widely applied chemical proteomic technologies, while delving into their latest applications in the target discovery of natural medicine. These endeavours demonstrate the potential of chemical proteomics for target discovery to supply dependable methodologies for the target elucidation of NPs.
由于天然产物(NPs)对蛋白质具有更好的选择性,全球一半以上的新型药物都直接或间接来自天然产物。传统药物通过各种 NPs 与药物靶点的结合发挥多种生物活性,这为药物开发带来了靶点发现的机遇。然而,检测 NPs 与靶点之间的结合关系仍然具有挑战性。化学蛋白质组学是化学、蛋白质组学、生物学和生物信息学的跨学科领域,已成为揭示药物与靶点相互作用的潜在方法。本综述总结了目前广泛应用的化学蛋白质组学技术的原理和特点,同时深入探讨了这些技术在天然药物靶点发现中的最新应用。这些努力证明了化学蛋白质组学在靶点发现方面的潜力,为阐明 NPs 靶点提供了可靠的方法。
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引用次数: 0
Pharmacological characterization of prostaglandin E2 EP2 and EP4 receptors in male rat locus coeruleus neurons ex vivo 雄性大鼠体外脑室神经元中前列腺素 E2 EP2 和 EP4 受体的药理学特征。
IF 5.3 2区 医学 Q1 PHARMACOLOGY & PHARMACY Pub Date : 2024-11-05 DOI: 10.1016/j.bcp.2024.116602
Amaia Nazabal, Aitziber Mendiguren, Joseba Pineda
The inflammatory mediator prostaglandin E2 (PGE2) binds to Gs-coupled EP2 and EP4 receptors. These receptors are located in the locus coeruleus (LC), the principal noradrenergic nucleus in the brain, but their functional role remains unknown. In this study, the PGE2 EP2 and EP4 receptors in LC cells from male rat brain slices were pharmacologically characterized by single-unit extracellular electrophysiology. The EP2 receptor agonists butaprost (0.01–10 μM) and treprostinil (0.03–10 µM) and the EP4 receptor agonists rivenprost (0.01 nM–1 µM) and TCS2510 (0.20 nM–2 µM) increased the firing rate of LC neurons in a concentration-dependent manner. The EP2 receptor antagonist PF-04418948 (10 nM) hindered the excitatory effect of butaprost and treprostinil while the EP4 receptor antagonist L-161,982 (30 and 300 nM) blocked the excitatory effect caused by rivenprost and TCS2510. The effects of butaprost and rivenprost were prevented by extracellular sodium replacement but were not modified by the protein kinase A (PKA) activator 8-Br-cAMP (1 mM) or the inhibitor H-89 (10 μM). However, the Gβγ subunit blocker gallein (20 μM) hindered the stimulatory effect of butaprost while the Gαs subunit inhibitor NF449 (10 µM) prevented that of rivenprost. Finally, rivenprost-induced stimulation (30 nM) was not occluded by butaprost (1 µM).
In conclusion, activation of EP2 and EP4 receptors excites LC noradrenergic neurons through sodium-dependent currents via different G protein subunits in male rat brain slices. EP2 and EP4 in the LC may constitute pharmacological targets for the treatment of pain, fever, drug addiction, anxiety and neuroinflammatory diseases.
炎症介质前列腺素 E2(PGE2)与 Gs 偶联的 EP2 和 EP4 受体结合。这些受体位于大脑中主要的去甲肾上腺素能神经核(LC),但其功能作用仍不清楚。本研究通过单体胞外电生理学对雄性大鼠脑切片 LC 细胞中的 PGE2 EP2 和 EP4 受体进行了药理学表征。EP2 受体激动剂丁前列素(0.01-10 μM)和曲普前列素(0.03-10 µM)以及 EP4 受体激动剂瑞文前列素(0.01 nM-1 µM)和 TCS2510(0.20 nM-2 µM)以浓度依赖的方式增加了 LC 神经元的发射率。EP2受体拮抗剂PF-04418948(10 nM)阻碍了丁前列素和曲前列素的兴奋作用,而EP4受体拮抗剂L-161982(30 nM和300 nM)阻断了里文前列素和TCS2510引起的兴奋作用。细胞外钠置换可阻止丁前列素和瑞文前列素的作用,但蛋白激酶 A(PKA)激活剂 8-Br-cAMP(1 mM)或抑制剂 H-89(10 μM)不会改变丁前列素和瑞文前列素的作用。然而,Gβγ亚基阻断剂加林(20 μM)阻碍了丁前列素的刺激作用,而Gαs亚基抑制剂NF449(10 μM)则阻止了利文前列素的刺激作用。最后,丁前列素(1 µM)不能阻止里文前列素诱导的刺激(30 nM)。总之,在雄性大鼠脑片中,EP2 和 EP4 受体通过不同的 G 蛋白亚基激活钠依赖性电流,从而兴奋 LC 去甲肾上腺素能神经元。LC中的EP2和EP4可能是治疗疼痛、发热、药物成瘾、焦虑和神经炎症性疾病的药理学靶点。
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引用次数: 0
Regulatory roles of histamine receptor in astrocytic glutamate clearance under conditions of increased glucose variability 组胺受体在葡萄糖变异性增加条件下对星形胶质细胞谷氨酸清除的调节作用
IF 5.3 2区 医学 Q1 PHARMACOLOGY & PHARMACY Pub Date : 2024-11-05 DOI: 10.1016/j.bcp.2024.116611
Yu Zhou , Wenhuo Xie , Chenghua Kong , Wei Luo , Hong Wei , Jiaping Zheng
In diabetic patients, repeated episodes of hypoglycemia can increase glucose variability (GV), which may lead to glutamate neurotoxicity in the brain and consequently affect cognitive functions. Astrocytes play a crucial role in regulating the balance of glutamate within the brain, and their function is influenced by the histamine receptor (HR) signaling pathway. However, the specific role of this mechanism under conditions of high GV is not yet clear. The results showed that increased GV resulted in decreased expression of HRs in mice hippocampus and astrocytes cultured in vitro. Additionally, a decrease in the expression of proteins related to glutamate metabolic clearance was observed, accompanied by a reduction in glutamate reuptake capacity. Notably, the intervention with histidine/histamine was able to reverse the above changes. Further mechanistic studies showed that inhibition of HRs that increased GV led to significant disturbances in astrocytic mitochondrial function. These abnormalities encompassed increased fragmentation morphology and the accumulation of reactive oxygen species, accompanied by decreased mitochondrial respiratory capacity and dysregulation of dynamics. Distinct HR subtypes exhibited variations in the modulation of mitochondrial function, with H3R demonstrating the most pronounced impact. The overexpression of H3R could enhance glutamate metabolic by reversing disturbances in mitochondrial dynamics. Therefore, this study suggests that H3R is able to maintain glutamate metabolic clearance capacity and exert neuroprotective effects in astrocytes that increased GV by regulating mitochondrial dynamic balance. This provides an important basis for potential therapeutic targets for diabetes-related cognitive dysfunction.
在糖尿病患者中,反复发作的低血糖会增加葡萄糖变异性(GV),从而可能导致大脑中的谷氨酸神经毒性,进而影响认知功能。星形胶质细胞在调节脑内谷氨酸平衡方面起着至关重要的作用,其功能受组胺受体(HR)信号通路的影响。然而,这一机制在高GV条件下的具体作用尚不清楚。研究结果表明,GV 的增加导致体外培养的小鼠海马和星形胶质细胞中 HRs 的表达减少。此外,还观察到与谷氨酸代谢清除相关的蛋白质表达减少,同时谷氨酸再摄取能力降低。值得注意的是,组氨酸/组胺的干预能够逆转上述变化。进一步的机理研究表明,抑制增加 GV 的 HRs 会导致星形胶质细胞线粒体功能的显著紊乱。这些异常包括碎片形态的增加和活性氧的积累,同时还伴随着线粒体呼吸能力的下降和动态失调。不同的 HR 亚型在线粒体功能调节方面表现出不同的差异,其中 H3R 的影响最为明显。H3R 的过表达可通过逆转线粒体动力学的紊乱来增强谷氨酸代谢。因此,本研究表明,H3R 能够维持谷氨酸代谢清除能力,并通过调节线粒体动态平衡,在 GV 增加的星形胶质细胞中发挥神经保护作用。这为糖尿病相关认知功能障碍的潜在治疗靶点提供了重要依据。
{"title":"Regulatory roles of histamine receptor in astrocytic glutamate clearance under conditions of increased glucose variability","authors":"Yu Zhou ,&nbsp;Wenhuo Xie ,&nbsp;Chenghua Kong ,&nbsp;Wei Luo ,&nbsp;Hong Wei ,&nbsp;Jiaping Zheng","doi":"10.1016/j.bcp.2024.116611","DOIUrl":"10.1016/j.bcp.2024.116611","url":null,"abstract":"<div><div>In diabetic patients, repeated episodes of hypoglycemia can increase glucose variability (GV), which may lead to glutamate neurotoxicity in the brain and consequently affect cognitive functions. Astrocytes play a crucial role in regulating the balance of glutamate within the brain, and their function is influenced by the histamine receptor (HR) signaling pathway. However, the specific role of this mechanism under conditions of high GV is not yet clear. The results showed that increased GV resulted in decreased expression of HRs in mice hippocampus and astrocytes cultured in vitro. Additionally, a decrease in the expression of proteins related to glutamate metabolic clearance was observed, accompanied by a reduction in glutamate reuptake capacity. Notably, the intervention with histidine/histamine was able to reverse the above changes. Further mechanistic studies showed that inhibition of HRs that increased GV led to significant disturbances in astrocytic mitochondrial function. These abnormalities encompassed increased fragmentation morphology and the accumulation of reactive oxygen species, accompanied by decreased mitochondrial respiratory capacity and dysregulation of dynamics. Distinct HR subtypes exhibited variations in the modulation of mitochondrial function, with H<sub>3</sub>R demonstrating the most pronounced impact. The overexpression of H<sub>3</sub>R could enhance glutamate metabolic by reversing disturbances in mitochondrial dynamics. Therefore, this study suggests that H<sub>3</sub>R is able to maintain glutamate metabolic clearance capacity and exert neuroprotective effects in astrocytes that increased GV by regulating mitochondrial dynamic balance. This provides an important basis for potential therapeutic targets for diabetes-related cognitive dysfunction.</div></div>","PeriodicalId":8806,"journal":{"name":"Biochemical pharmacology","volume":"230 ","pages":"Article 116611"},"PeriodicalIF":5.3,"publicationDate":"2024-11-05","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142603100","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
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Biochemical pharmacology
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