Cellular signalling最新文献_第7页

Puerarin protects against renal ischemia-reperfusion injury by restoring mitochondrial function and modulating the PI3K/AKT/NF-k B pathway and suppressing inflammatory responses 葛根素通过恢复线粒体功能、调节PI3K/AKT/NF-k - B通路和抑制炎症反应来预防肾缺血再灌注损伤

IF 3.7 2区生物学 Q2 CELL BIOLOGY

Cellular signalling

Pub Date : 2025-12-31 DOI: 10.1016/j.cellsig.2025.112350

Kangyu Wang , Changhong Xu , Hao Wang , Yalong Zhang , Zijian Zhang , Rui Yan , Li Wang , Jianwei Yang , Jiangwei Man , Li Yang

Background

Renal ischemia–reperfusion injury (RIRI) is a major cause of perioperative acute kidney injury and a driver of chronic kidney disease, in which mitochondrial dysfunction and inflammatory activation create a vicious cycle of oxidative stress, inflammation, and tubular cell death. Puerarin, a natural isoflavone with established cardio- and neuroprotective effects, has not been fully evaluated as a modulator of the mitochondrial–immune axis in RIRI.

Methods

Network pharmacology and RNA sequencing of mouse kidneys with bilateral RIRI were integrated to identify shared puerarin–RIRI targets and enriched pathways. Molecular docking and 100-ns molecular dynamics simulations were performed to assess puerarin–PI3K binding. In vivo, C57BL/6 mice underwent 45-min bilateral renal ischemia and 24-h reperfusion with or without puerarin pretreatment and/or the PI3K inhibitor LY294002. In vitro, HK-2 cells were subjected to hypoxia/reoxygenation. Renal injury, apoptosis, oxidative stress, inflammatory cytokines, mitochondrial ultrastructure, membrane potential, and mitochondrial dynamics proteins (Drp1, Opa1, Mfn2) were evaluated.

Results

Network pharmacology and transcriptomics consistently pointed to the PI3K/AKT/NF-κB pathway, which was enriched in oxidative stress, apoptosis, inflammatory signaling, and mitochondrial processes. In both RIRI mice and HR-treated HK-2 cells, puerarin improved renal histology and function, reduced tubular apoptosis and IL-6/IL-1β/TNF-α levels, restored SOD and GSH-Px, lowered ROS and MDA, and preserved mitochondrial membrane potential and ultrastructure. These benefits were accompanied by activation of PI3K/AKT signaling and attenuation of NF-κB, together with normalization of mitochondrial fission–fusion (decreased Drp1, increased Opa1/Mfn2), whereas LY294002 markedly blunted puerarin-induced improvements in mitochondrial dynamics, membrane potential, and injury indices.

Conclusion

Puerarin ameliorates RIRI by activating PI3K/AKT signaling, restoring mitochondrial homeostasis, and attenuating NF-κB–linked inflammatory and apoptotic responses. These findings support puerarin as a mitochondria- and immune-targeted candidate for perioperative renal protection and justify further translational evaluation.

肾缺血再灌注损伤（RIRI）是围手术期急性肾损伤的主要原因，也是慢性肾脏疾病的驱动因素，其中线粒体功能障碍和炎症激活造成氧化应激、炎症和小管细胞死亡的恶性循环。葛根素是一种天然异黄酮，具有心脏和神经保护作用，但在RIRI中作为线粒体-免疫轴的调节剂尚未得到充分评估。方法结合网络药理学和双侧RIRI小鼠肾脏的RNA测序，确定葛根素- RIRI的共同靶点和富集途径。通过分子对接和100-ns分子动力学模拟来评估葛根素- pi3k的结合。在体内，C57BL/6小鼠在葛根素预处理和/或PI3K抑制剂LY294002或不加葛根素预处理和/或不加PI3K抑制剂LY294002的情况下进行45分钟双侧肾缺血和24小时再灌注。在体外，HK-2细胞经历缺氧/再氧化。评估肾损伤、细胞凋亡、氧化应激、炎症因子、线粒体超微结构、膜电位和线粒体动力学蛋白（Drp1、Opa1、Mfn2）。结果网络药理学和转录组学一致指向PI3K/AKT/NF-κB通路，该通路在氧化应激、细胞凋亡、炎症信号和线粒体过程中富集。在RIRI小鼠和hr处理的HK-2细胞中，葛根素改善肾组织和功能，减少肾小管凋亡和IL-6/IL-1β/TNF-α水平，恢复SOD和GSH-Px，降低ROS和MDA，保存线粒体膜电位和超微结构。这些益处伴随着PI3K/AKT信号的激活和NF-κB的衰减，以及线粒体裂变融合的正常化（Drp1降低，Opa1/Mfn2增加），而LY294002明显减弱了葛根素诱导的线粒体动力学、膜电位和损伤指标的改善。结论葛根素通过激活PI3K/AKT信号，恢复线粒体稳态，减弱NF-κ b相关的炎症和凋亡反应，改善RIRI。这些发现支持葛根素作为围手术期肾脏保护的线粒体和免疫靶向候选物，并证明进一步的翻译评价是合理的。

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引用次数: 0

Selenoprotein P drives glioma malignancy via LRP8-dependent activation of Wnt/β-catenin signaling 硒蛋白P通过lrp8依赖的Wnt/β-catenin信号激活驱动胶质瘤恶性。

IF 3.7 2区生物学 Q2 CELL BIOLOGY

Cellular signalling

Pub Date : 2025-12-29 DOI: 10.1016/j.cellsig.2025.112341

Xiaoke Wu , Yao Zhang , Guang Rong , Zixiao Zhang , Jieyu Jin , Junchao Feng , Haiyang Wu , Yin Wang

Glioma is a highly aggressive central nervous system malignancy with poor prognosis and limited therapeutic options. Selenoprotein P (SeP) plays key roles in the nervous system, but its function in glioma remains unclear. Analysis of TCGA and CGGA datasets revealed that SeP is highly expressed in glioma tissues and correlates with poor patient survival. In vitro, SeP knockdown inhibited glioma cell proliferation, migration, invasion, and epithelial–mesenchymal transition (EMT), induced mitochondrial-dependent apoptosis, and reduced nuclear β-catenin and downstream targets Cyclin D1 and c-Myc, whereas SeP overexpression produced opposite effects. Mechanistically, SeP directly binds to low-density lipoprotein receptor-related protein 8 (LRP8) to activate Wnt/β-catenin signaling. Rescue experiments showed that LRP8 overexpression or Wnt agonist treatment reversed the inhibitory effects of SeP knockdown, while LRP8 knockdown or Wnt inhibition blocked SeP-induced malignancy. In subcutaneous xenograft models, SeP silencing suppressed tumor growth, decreased Ki-67 proliferation, and increased Caspase-3–mediated apoptosis, whereas LRP8 overexpression restored tumor growth and Wnt/β-catenin activity. Collectively, these results demonstrate that SeP promotes glioma malignancy via an LRP8-dependent activation of Wnt/β-catenin signaling and suggest the SeP–LRP8–Wnt/β-catenin axis as a potential therapeutic target.

胶质瘤是一种高度侵袭性的中枢神经系统恶性肿瘤，预后差，治疗选择有限。硒蛋白P （Selenoprotein P, SeP）在神经系统中起重要作用，但其在神经胶质瘤中的功能尚不清楚。对TCGA和CGGA数据集的分析显示，SeP在胶质瘤组织中高表达，并与患者生存率低相关。在体外，SeP敲低可抑制胶质瘤细胞的增殖、迁移、侵袭和上皮-间质转化（EMT），诱导线粒体依赖性细胞凋亡，降低核β-catenin及其下游靶点Cyclin D1和c-Myc，而SeP过表达则产生相反的作用。在机制上，SeP直接结合低密度脂蛋白受体相关蛋白8 （LRP8）激活Wnt/β-catenin信号。救援实验表明，LRP8过表达或Wnt激动剂治疗可逆转SeP敲低的抑制作用，而LRP8敲低或Wnt抑制可阻断SeP诱导的恶性肿瘤。在皮下异种移植物模型中，SeP沉默抑制肿瘤生长，降低Ki-67增殖，增加caspase -3介导的细胞凋亡，而LRP8过表达恢复肿瘤生长和Wnt/β-catenin活性。总之，这些结果表明SeP通过lrp8依赖性的Wnt/β-catenin信号激活促进胶质瘤恶性肿瘤，并提示SeP- lrp8 -Wnt/β-catenin轴是一个潜在的治疗靶点。

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引用次数: 0

ISG15 dysregulates endoplasmic reticulum-mitochondrial contacts and calcium homeostasis in Ataxia telangiectasia 在共济失调毛细血管扩张中，ISG15异常调节内质网-线粒体接触和钙稳态。

IF 3.7 2区生物学 Q2 CELL BIOLOGY

Cellular signalling

Pub Date : 2025-12-29 DOI: 10.1016/j.cellsig.2025.112347

Oygul Mirzalieva, Ryan E. Reed, Arthur L. Haas, Meredith A. Juncker , Patrick Logarbo, Jennifer M. Klein, David Worthylake, Shyamal D. Desai

Dysregulation of endoplasmic reticulum and mitochondrial (ER:Mit) contacts and mitochondrial calcium (mitCa²⁺) homeostasis are found in several neurodegenerative disorders, including Ataxia Telangiectasia (A-T). However, the cellular basis of these defects remains unclear. Previously, we demonstrated that the aberrantly elevated Interferon-Stimulated Gene 15 (ISG15) pathway inhibits protein polyubiquitylation, its dependent protein turnover, and mitophagy pathways in A-T. Literature indicates that silencing of mitochondrial ubiquitin ligase 1 (MUL1) stabilizes mitofusin2 (MFN2) and attenuates mitCa²⁺ uptake from ER to Mit (mitCa²⁺_influx) in primary neurons. We have replicated these findings in apparently healthy fibroblasts. We hypothesized that elevated ISG15 may inhibit ubiquitin-dependent MUL1-mediated degradation of MFN2 and dysregulate ER:Mit contacts and mitCa²⁺ homeostasis in A-T fibroblasts. Concurrently, MFN2 is stabilized in A-T, MUL1-silenced A-T, MUL1/ISG15-silenced A-T vs ISG15-silenced A-T fibroblasts. Moreover, the number of ER:Mit contacts is increased in A-T vs ISG15-silenced A-T fibroblasts. Notably, mitCa²⁺_efflux is significantly attenuated in A-T vs ISG15-silenced A-T fibroblasts in which mitCa²⁺_efflux is restored to levels comparable to those observed in normal fibroblasts. The mitCa²⁺_efflux remains attenuated in MUL1 and MUL1/ISG15-silenced A-T fibroblasts. We conclude that ISG15 impairs MUL1/MFN2-mediated regulation of ER:Mit contacts and attenuates mitCa²⁺_efflux, which may, in turn, cause Ca²⁺ overload-mediated mitochondrial damage in A-T. These findings suggest that ISG15 silencers may correct mitochondrial abnormalities and improve mitochondrial health in A-T patients and in those with other neurodegenerative disorders in which ISG15 is elevated, such as ALS.

内质网和线粒体（ER:Mit）接触和线粒体钙（mitCa2+）稳态失调在一些神经退行性疾病中被发现，包括共济失调毛细血管扩张症（A-T）。然而，这些缺陷的细胞基础仍不清楚。先前，我们证明了异常升高的干扰素刺激基因15 （ISG15）通路抑制蛋白多泛素化，其依赖的蛋白转换和线粒体自噬通路。文献表明，线粒体泛素连接酶1 （MUL1）的沉默稳定了线粒体分裂素2 (MFN2)，并减弱了初级神经元从ER到Mit的mitCa2+摄取（mitCa2+内流）。我们在表面健康的成纤维细胞中重复了这些发现。我们假设升高的ISG15可能抑制泛素依赖的mul1介导的MFN2降解，并在A-T成纤维细胞中失调ER:Mit接触和mitCa2+稳态。同时，MFN2在A-T、MUL1沉默的A-T、MUL1/ isg15沉默的A-T和isg15沉默的A-T成纤维细胞中是稳定的。此外，与isg15沉默的A-T成纤维细胞相比，A-T中ER:Mit接触的数量增加。值得注意的是，与isg15沉默的A-T成纤维细胞相比，A-T中mitCa2+外排明显减弱，其中mitCa2+外排恢复到与正常成纤维细胞相当的水平。mitCa2+外排在MUL1和MUL1/ isg15沉默的A-T成纤维细胞中仍然减弱。我们得出的结论是，ISG15损害了MUL1/ mfn2介导的ER:Mit接触和减弱mitCa2+外排的调节，这可能反过来导致Ca2+超载介导的线粒体损伤。这些发现表明，ISG15沉默剂可能纠正线粒体异常并改善A-T患者和其他ISG15升高的神经退行性疾病（如ALS）患者的线粒体健康。

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引用次数: 0

Targeting USP28 inhibits clear cell renal cell carcinoma growth 靶向USP28抑制透明细胞肾细胞癌生长。

IF 3.7 2区生物学 Q2 CELL BIOLOGY

Cellular signalling

Pub Date : 2025-12-26 DOI: 10.1016/j.cellsig.2025.112344

Ying Ren , Yunfang Yang , Xiaodan Zhu , Lingyi Li , Yaoyao Pan , Hong Sun , Qian Fang , Hui Xiong , Dong Guo , Yongzhan Sun , Hailong Li , Junqi Wang

Clear cell renal cell carcinoma (ccRCC), the most common type of kidney cancer, remains challenging to treat due to the lack of effective targeted therapies. Although c-Myc is frequently overexpressed in ccRCC, the mechanisms governing its stability are not well understood. Here, we identify the deubiquitinating enzyme USP28 as a key regulator of c-Myc protein stability in ccRCC. USP28 interacts with c-Myc, removes K48-linked polyubiquitin chains, and thereby prevents its proteasomal degradation, leading to c-Myc stabilization in ccRCC cells. Genetic or pharmacological inhibition of USP28 significantly reduces c-Myc expression, impairs ccRCC cell proliferation in vitro, and suppresses tumor growth in vivo. Tumors with high c-Myc expression exhibit heightened sensitivity to USP28 inhibition, underscoring the therapeutic potential of targeting this axis. Collectively, our findings position USP28 as a promising therapeutic target for ccRCC.

透明细胞肾细胞癌（ccRCC）是最常见的肾癌类型，由于缺乏有效的靶向治疗，治疗仍然具有挑战性。尽管c-Myc在ccRCC中经常过表达，但控制其稳定性的机制尚不清楚。在这里，我们发现去泛素化酶USP28是ccRCC中c-Myc蛋白稳定性的关键调节因子。USP28与c-Myc相互作用，去除k48连接的多泛素链，从而阻止其蛋白酶体降解，导致ccRCC细胞中c-Myc稳定。遗传或药理学抑制USP28可显著降低c-Myc表达，损害体外ccRCC细胞增殖，抑制体内肿瘤生长。高c-Myc表达的肿瘤表现出对USP28抑制的高度敏感性，强调了靶向该轴的治疗潜力。总的来说，我们的发现将USP28定位为ccRCC的一个有希望的治疗靶点。

引用次数: 0

BDH1 regulates cardiomyocyte apoptosis and diabetic cardiomyopathy by modulating mitochondrial dynamics and attenuating oxidative stress BDH1通过调节线粒体动力学和减轻氧化应激来调节心肌细胞凋亡和糖尿病性心肌病。

IF 3.7 2区生物学 Q2 CELL BIOLOGY

Cellular signalling

Pub Date : 2025-12-26 DOI: 10.1016/j.cellsig.2025.112345

Yuxuan Shi , Guoqiang Xu , Shuang Liu , Xin Liu , Wendi Chen , Guolong Wang , Yongzhi Cao , Yunna Ning , Yanfei Jia , Yueran Zhao

Diabetic cardiomyopathy (DCM) is a significant cardiovascular consequence of diabetes, marked by increasing myocardial dysfunction that culminates in heart failure. Current treatments prioritize glucose control but frequently neglect the fundamental metabolic dysfunction and excessive cardiomyocyte death that propel disease progression. Recent data indicates that β-hydroxybutyrate dehydrogenase 1 (BDH1), the rate-limiting enzyme in ketolysis may serve a protective function by supplying alternative energy substrates and regulating apoptosis. This study demonstrates that BDH1 overexpression in H9c2 and AC16 cardiomyocytes treated with high glucose and palmitate dramatically decreases apoptosis. In a streptozotocin-induced diabetic mouse model, cardiac-specific BDH1 overexpression via AAV9 delivery maintained cardiac structure and function by inhibiting apoptosis through two primary mechanisms: (1) interaction with KIF1Bβ to modulate mitochondrial dynamics, and (2) suppression of ROS-mediated apoptosome formation (Cytc/Apaf-1 complex). These findings designate BDH1 as an innovative therapeutic target for DCM by illustrating its dual function in metabolic regulation and apoptosis inhibition, while recognizing ROS and KIF1Bβ as pivotal mediators of DCM pathogenesis.

糖尿病性心肌病（DCM）是糖尿病的重要心血管后果，其特征是心肌功能障碍增加，最终导致心力衰竭。目前的治疗优先考虑血糖控制，但经常忽视推动疾病进展的基本代谢功能障碍和过度的心肌细胞死亡。最近的研究表明，酮解过程中的限速酶β-羟基丁酸脱氢酶1 （BDH1）可能通过提供替代能量底物和调节细胞凋亡而发挥保护作用。本研究表明，BDH1在高糖和棕榈酸盐处理的H9c2和AC16心肌细胞中过表达可显著减少细胞凋亡。在链脲霉素诱导的糖尿病小鼠模型中，通过AAV9传递心脏特异性BDH1过表达，通过两种主要机制通过抑制凋亡维持心脏结构和功能：(1)与KIF1Bβ相互作用调节线粒体动力学，(2)抑制ros介导的凋亡形成（Cytc/Apaf-1复合物）。这些发现表明BDH1具有代谢调节和细胞凋亡抑制的双重功能，是DCM的创新治疗靶点，同时认识到ROS和KIF1Bβ是DCM发病机制的关键介质。

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引用次数: 0

Cornus officinalis total glycosides modulate inflammatory response and inhibit the JAK1/STAT3 pathway within a preclinical rat model of cardiac ischemia/reperfusion-induced injury 山茱萸总苷在临床前大鼠心脏缺血/再灌注损伤模型中调节炎症反应并抑制JAK1/STAT3通路

IF 3.7 2区生物学 Q2 CELL BIOLOGY

Cellular signalling

Pub Date : 2025-12-26 DOI: 10.1016/j.cellsig.2025.112346

Ke Chen , Yujie Lin , Liudan Chen, Jianjun Li, Aizhen Pan

Objective

Cardiac ischemia/reperfusion (I/R) injury is characterized by excessive inflammation and aberrant activation of intracellular signaling pathways, including JAK/STAT. Cornus officinalis total glycosides (COTG) possess anti-inflammatory properties, but their role in myocardial I/R injury remains incompletely defined. This study aimed to investigate the cardioprotective effects of COTG against I/R -induced injury and clarify its pathway-dependent mechanism involving JAK1/STAT3 signaling.

Methods

A myocardial I/R injury model was established in male Sprague–Dawley rats, which were randomly divided into Sham control, I/R control, and COTG-treated groups (200, 400, and 600 mg/kg). Cardiac function was evaluated by echocardiography, and myocardial histopathology was assessed by hematoxylin and eosin staining. Serum myocardial injury markers were measured by ELISA. Inflammatory cytokine expression and macrophage polarization were analyzed using qRT-PCR and immunofluorescence, respectively. JAK1/2 and STAT3 phosphorylation was detected by Western blotting. An in vitro hypoxia/reoxygenation (H/R) model combined with a pharmacological JAK inhibitor was further employed to verify pathway dependency.

Results

COTG treatment significantly improved cardiac function, as evidenced by increased left ventricular ejection fraction and fractional shortening and reduced ventricular dilation. Histological damage and serum levels of cardiac troponin I, cardiac troponin T, and creatine kinase-MB were markedly attenuated. COTG suppressed pro-inflammatory markers (iNOS, IL-1β, IL-6) while enhancing anti-inflammatory mediators (Arg-1, IL-10). Mechanistically, COTG dose-dependently inhibited JAK1, JAK2, and STAT3 phosphorylation without altering total protein levels. Pharmacological inhibition experiments confirmed that JAK/STAT suppression by COTG was pathway-dependent. Moreover, COTG reduced myocardial apoptosis by increasing Bcl-2 and decreasing Bax and cleaved caspase-3 expression.

Conclusion

COTG protects against myocardial I/R injury by pathway-dependent inhibition of JAK/STAT signaling, modulation of inflammatory responses, and attenuation of cardiomyocyte apoptosis, highlighting its therapeutic potential for ischemic heart disease.

目的：心脏缺血再灌注（I/R）损伤以过度炎症和细胞内信号通路异常激活为特征，包括JAK/STAT。山茱萸总苷（COTG）具有抗炎作用，但其在心肌I/R损伤中的作用尚未完全确定。本研究旨在探讨COTG对I/R诱导损伤的心脏保护作用，并阐明其涉及JAK1/STAT3信号通路依赖的机制。方法：建立雄性Sprague-Dawley大鼠心肌I/R损伤模型，随机分为Sham对照组、I/R对照组和cotg治疗组（200、400、600 mg/kg）。超声心动图评价心功能，苏木精和伊红染色评价心肌组织病理学。采用ELISA法测定血清心肌损伤标志物。分别用qRT-PCR和免疫荧光分析炎症细胞因子表达和巨噬细胞极化。Western blotting检测JAK1/2和STAT3磷酸化水平。进一步采用体外缺氧/再氧化（H/R）模型联合药理JAK抑制剂验证通路依赖性。结果：COTG治疗可显著改善心功能，左心室射血分数和缩短分数增加，心室舒张降低。组织学损伤和心肌肌钙蛋白I、心肌肌钙蛋白T和肌酸激酶- mb的血清水平明显减弱。COTG抑制促炎标志物（iNOS, IL-1β, IL-6），增强抗炎介质（Arg-1, IL-10）。在机制上，COTG剂量依赖性地抑制JAK1、JAK2和STAT3磷酸化，而不改变总蛋白水平。药理抑制实验证实，COTG对JAK/STAT的抑制具有通路依赖性。此外，COTG通过增加Bcl-2、降低Bax和cleaved caspase-3的表达来减少心肌凋亡。结论：COTG通过通路依赖性抑制JAK/STAT信号通路、调节炎症反应和抑制心肌细胞凋亡来保护心肌I/R损伤，突出了其对缺血性心脏病的治疗潜力。

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引用次数: 0

Hsp70/CHIP E3 ligase complex triggers K149-linked ubiquitination and degradation of BEST1 mutants p.P233L and p.P346H, impairing chloride channel function and retinal integrity Hsp70/CHIP E3连接酶复合物触发k149连接的泛素化和降解BEST1突变体p.P233L和p.P346H，损害氯通道功能和视网膜完整性。

IF 3.7 2区生物学 Q2 CELL BIOLOGY

Cellular signalling

Pub Date : 2025-12-26 DOI: 10.1016/j.cellsig.2025.112343

Zhongxue Zhou , Hongxia Tian , Ning Ma , Wen Fang , Qingxia Lu , Huiyang , HaiyingTian , Yu Tang , Ling Tian , Xu Jia , Yuanju Zhang , Yuqian Li , Xiaoyan Zhu , Qiao Yu , Ding'an Zhou

The mechanisms underlying ubiquitination-mediated degradation of bestrophinopathy-causing mutants and their in vivo effects on retinal pigment epithelium (RPE) localization and retinal structure remain poorly understood. Furthermore, the upstream signaling cascade that induces degradation of these mutants and the detailed ubiquitination mechanism are unknown. Here, we report a c.1037C > A (p.P346H) mutation and a c.698C > T (p.P233L) mutation that co-segregate with the phenotypes of pedigrees affected by RP50 and Best vitelliform macular dystrophy (BVMD), respectively. The BEST1 mutants p.P233L and p.P346H reduced chloride channel activity and induced mislocalization of bestrophin-1 in polarized MDCK II cells, significantly affecting the channel activity of wild-type bestrophin-1. Lys149 was identified as the site responsible for ubiquitination of p.P346H- and p.P233L-bestrophin-1, mediated by Hsp70 and the C-terminal Hsp70-interacting protein (CHIP). Mutant bestrophin-1 proteins p.P346H and p.P233L undergo ubiquitination and degradation, preventing their localization to the cell membrane of MDCK II cells and the RPE of zebrafish, thereby reducing chloride channel activity. Mislocalization of mutant bestrophin-1 to the RPE impaired the multicellular layered structure of the retina. Our study reveals a ubiquitination signaling pathway mediated by Hsp70 and CHIP that depends on Lys149 of bestrophin-1. Aberrant activation of this pathway leads to loss of function in the p.P233L and p.P346H mutants and triggers retinopathy.

泛素化介导的白视病突变体降解的机制及其对视网膜色素上皮（RPE）定位和视网膜结构的体内影响尚不清楚。此外，诱导这些突变体降解的上游信号级联和详细的泛素化机制尚不清楚。在这里，我们报道了一个c.1037C > a （p.P346H）突变和一个c.698C > T （p.P233L）突变，它们分别与受RP50和BVMD影响的家系表型共分离。BEST1突变体p.P233L和p.P346H降低了MDCK II细胞中氯离子通道的活性，诱导了bestrophin-1的错定位，显著影响了野生型bestrophin-1的通道活性。Lys149被鉴定为由Hsp70和c端Hsp70相互作用蛋白（CHIP）介导的p.P346H-和p.p 233l - strophin-1泛素化的位点。突变的bestrophin-1蛋白p.P346H和p.P233L发生泛素化和降解，阻止其定位于MDCK II细胞的细胞膜和斑马鱼的RPE，从而降低氯离子通道活性。突变的strophin-1在RPE上的错误定位损害了视网膜的多细胞分层结构。我们的研究揭示了一个由Hsp70和CHIP介导的泛素化信号通路，它依赖于bestrophin-1的Lys149。该通路的异常激活导致p.P233L和p.P346H突变体功能丧失，并引发视网膜病变。

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引用次数: 0

The anti-aging drug Senolytics alleviates vascular endothelial cell senescence by antagonizing the JPX-BRD4-p65 complex 抗衰老药物Senolytics通过拮抗JPX-BRD4-p65复合物缓解血管内皮细胞衰老。

IF 3.7 2区生物学 Q2 CELL BIOLOGY

Cellular signalling

Pub Date : 2025-12-23 DOI: 10.1016/j.cellsig.2025.112342

Kaili Mao , Wensi Zhu , Huayu Sun , Hongliang Zheng , Zhenling Fu

Senescence of vascular endothelial cells (ECs) and smooth muscle cells is pivotal in vascular aging. While the long non-coding RNA JPX has been shown to induce smooth muscle cell senescence, its role in EC senescence remains unknown. Senolytics (dasatinib + quercetin, D + Q) can eliminate senescent cells and attenuate the senescence-associated secretory phenotype (SASP), but their mechanisms in ECs are unclear. Here, we investigated the interplay between Senolytics and JPX in human umbilical vein endothelial cells (HUVECs). Using a lipopolysaccharide (LPS)-induced senescence model, we found that Senolytics alleviated endothelial senescence while suppressing the upregulation of JPX observed in senescent HUVECs. Functional assays showed that JPX knockdown inhibited, whereas JPX overexpression aggravated, EC senescence. Mechanistically, JPX interacted with bromodomain-containing protein 4 (BRD4) and transcription factor p65 to form a JPX-BRD4-p65 complex that promoted SASP expression. In vivo, Senolytics disrupted this complex and effectively mitigated endothelial senescence. Collectively, our study identifies JPX as a key regulator of vascular endothelial senescence and reveals that Senolytics exert anti-senescent effects by antagonizing the JPX-BRD4-p65 complex, providing novel mechanistic insights and potential therapeutic targets for vascular aging–related diseases.

血管内皮细胞（ECs）和平滑肌细胞的衰老是血管衰老的关键。虽然长链非编码RNA JPX已被证明可诱导平滑肌细胞衰老，但其在EC衰老中的作用尚不清楚。抗衰老药物（达沙替尼+槲皮素，D + Q）可以消除衰老细胞并减轻衰老相关分泌表型（SASP），但其在ECs中的机制尚不清楚。在这里，我们研究了Senolytics和JPX在人脐静脉内皮细胞（HUVECs）中的相互作用。通过脂多糖（LPS）诱导的衰老模型，我们发现Senolytics可以缓解内皮细胞的衰老，同时抑制衰老HUVECs中JPX的上调。功能分析表明，JPX的下调抑制了EC的衰老，而JPX的过表达则加剧了EC的衰老。机制上，JPX与含溴结构域蛋白4 （BRD4）和转录因子p65相互作用，形成JPX-BRD4-p65复合物，促进SASP的表达。在体内，Senolytics破坏了这个复合体，有效地减轻了内皮细胞的衰老。总之，我们的研究确定了JPX是血管内皮衰老的关键调节因子，并揭示了Senolytics通过拮抗JPX- brd4 -p65复合物发挥抗衰老作用，为血管衰老相关疾病提供了新的机制见解和潜在的治疗靶点。

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引用次数: 0

Roxadustat induces erythroid differentiation of erythroleukemia cells through the hypoxia inducible factor-α/GATA binding protein 1 axis 罗沙司他通过缺氧诱导因子-α / GATA结合蛋白1轴诱导红系白血病细胞分化。

IF 3.7 2区生物学 Q2 CELL BIOLOGY

Cellular signalling

Pub Date : 2025-12-22 DOI: 10.1016/j.cellsig.2025.112340

Liqing Yang , Zhehan Luo , Shuang Wang , Jia Lin , Yuanzhong Chen , Yong Wu

Background

Acute erythroleukemia (AEL) is a rare type of acute myeloid leukemia characterized by severe anemia. Roxadustat is a marketed first-generation hypoxia inducible factor-proline hydroxylase inhibitor (HIF-PHI), currently widely used in anemia associated with chronic kidney disease. Studies on roxadustat in anemia related to myeloid neoplasms remains limited.

Methods

Erythroid differentiation levels of HEL and K562 cells after roxadustat treatment were assessed via flow cytometry and benzidine staining. Real timequantitative PCR and western blot were performed to detect mRNA and protein expression levels of erythroid differentiation-related genes. Small interfering RNA (siRNA) was used to knock down HIF-1α or HIF-2α to elucidate the primary target of roxadustat-induced erythropoiesis. To study the role of GATA binding protein 1 (GATA1) in roxadustat-induced erythropoiesis, CRISPR/Cas9-mediated GATA1 knockout cell lines were established. Chromatin immunoprecipitation-PCR and dualluciferase reporter assay were conducted to determine the interaction between HIF-α and GATA1.

Results

Roxadustat significantly promoted erythroid differentiation in HEL and K562 cells. Knockdown of HIF-2α rather than HIF-1α, and knockout of GATA1 both effectively inhibited this effect. Both HIF-1α and HIF-2α are responsible for the direct upregulation of GATA1 expression under hypoxia, with HIF-1α as the dominant transcription factor. Despite potentially different roles of HIF-1α and HIF-2α in leukemogenesis, roxadustat combined with ruxolitinib or imatinib exhibited synergistic anti-leukemia effect in HEL and K562 cells, respectively.

Conclusions

Roxadustat induces erythropoiesis in erythroleukemia cells primarily by targeting HIF-2α and was partially GATA1-dependent. Besides upregulating erythropoietin, our study revealed the HIF-α/GATA1 axis as another critical regulatory mechanism for hypoxia-driven erythropoiesis.

背景：急性红细胞白血病（AEL）是一种罕见的急性髓系白血病，以严重贫血为特征。Roxadustat是已上市的第一代缺氧诱导因子-脯氨酸羟化酶抑制剂（HIF-PHI），目前广泛用于慢性肾脏疾病相关贫血。关于罗沙司他治疗髓系肿瘤相关性贫血的研究仍然有限。方法：采用流式细胞术和联苯胺染色检测罗沙他他治疗后HEL和K562细胞的红系分化水平。采用实时定量PCR和western blot检测红系分化相关基因mRNA和蛋白的表达水平。利用小干扰RNA （siRNA）敲除细胞中HIF-1α或HIF-2α的表达，以阐明罗昔杜司他诱导红细胞生成的主要靶点。为了研究GATA结合蛋白1 （GATA1）在罗昔杜司他诱导的红细胞生成中的作用，我们建立了CRISPR/ cas9介导的GATA1敲除细胞系。采用染色质免疫沉淀- pcr和双荧光素酶报告基因法检测HIF-α与GATA1的相互作用。结果：罗沙司他显著促进HEL和K562细胞红细胞分化。敲除HIF-2α而非HIF-1α和敲除GATA1均能有效抑制这种作用。缺氧条件下，HIF-1α和HIF-2α都直接上调GATA1的表达，HIF-1α是主要的转录因子。尽管HIF-1α和HIF-2α在白血病发生中的作用可能不同，但罗胥达联合鲁索利替尼或伊马替尼在HEL和K562细胞中均表现出协同抗白血病作用。结论：罗沙司他主要通过靶向HIF-2α诱导红细胞生成，部分依赖于gata1。除了促红细胞生成素的上调，我们的研究揭示了HIF-α/GATA1轴是缺氧驱动的红细胞生成的另一个关键调节机制。

{"title":"Roxadustat induces erythroid differentiation of erythroleukemia cells through the hypoxia inducible factor-α/GATA binding protein 1 axis","authors":"Liqing Yang , Zhehan Luo , Shuang Wang , Jia Lin , Yuanzhong Chen , Yong Wu","doi":"10.1016/j.cellsig.2025.112340","DOIUrl":"10.1016/j.cellsig.2025.112340","url":null,"abstract":"<div><h3>Background</h3><div>Acute erythroleukemia (AEL) is a rare type of acute myeloid leukemia characterized by severe anemia. Roxadustat is a marketed first-generation hypoxia inducible factor-proline hydroxylase inhibitor (HIF-PHI), currently widely used in anemia associated with chronic kidney disease. Studies on roxadustat in anemia related to myeloid neoplasms remains limited.</div></div><div><h3>Methods</h3><div>Erythroid differentiation levels of HEL and K562 cells after roxadustat treatment were assessed via flow cytometry and benzidine staining. Real timequantitative PCR and western blot were performed to detect mRNA and protein expression levels of erythroid differentiation-related genes. Small interfering RNA (siRNA) was used to knock down <em>HIF-1α</em> or <em>HIF-2α</em> to elucidate the primary target of roxadustat-induced erythropoiesis. To study the role of GATA binding protein 1 (GATA1) in roxadustat-induced erythropoiesis, CRISPR/Cas9-mediated <em>GATA1</em> knockout cell lines were established. Chromatin immunoprecipitation-PCR and dualluciferase reporter assay were conducted to determine the interaction between HIF-α and GATA1.</div></div><div><h3>Results</h3><div>Roxadustat significantly promoted erythroid differentiation in HEL and K562 cells. Knockdown of <em>HIF-2α</em> rather than <em>HIF-1α</em>, and knockout of <em>GATA1</em> both effectively inhibited this effect. Both HIF-1α and HIF-2α are responsible for the direct upregulation of GATA1 expression under hypoxia, with HIF-1α as the dominant transcription factor. Despite potentially different roles of HIF-1α and HIF-2α in leukemogenesis, roxadustat combined with ruxolitinib or imatinib exhibited synergistic anti-leukemia effect in HEL and K562 cells, respectively.</div></div><div><h3>Conclusions</h3><div>Roxadustat induces erythropoiesis in erythroleukemia cells primarily by targeting HIF-2α and was partially GATA1-dependent. Besides upregulating erythropoietin, our study revealed the HIF-α/GATA1 axis as another critical regulatory mechanism for hypoxia-driven erythropoiesis.</div></div>","PeriodicalId":9902,"journal":{"name":"Cellular signalling","volume":"139 ","pages":"Article 112340"},"PeriodicalIF":3.7,"publicationDate":"2025-12-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145826989","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Bioinformatic analysis identifies RGS10 as a modulator of BV-2 microglia migration 生物信息学分析表明RGS10是BV-2小胶质细胞迁移的调节剂。

IF 3.7 2区生物学 Q2 CELL BIOLOGY

Cellular signalling

Pub Date : 2025-12-20 DOI: 10.1016/j.cellsig.2025.112338

Shwetal Talele , Menbere Wendimu , Shelley B. Hooks , Benita Sjögren

Regulator of G protein signaling 10 (RGS10) is a GTPase activating protein, selective for Gα_i, that has been proposed to play a role in suppressing microglia-driven neuroinflammation. RGS10 expression in microglia is suppressed by inflammatory stimuli and aging, and loss of RGS10 is associated with increased cytokine expression and neurodegeneration. Conversely, RGS10 overexpression provides protection against inflammatory stimuli in rodent models, however the mechanisms by which RGS10 exerts this protective effect are unknown. To understand the neuroprotective functions of RGS10 in microglia, we completed RNA-Seq analysis in the murine microglial cell line BV-2 with intact (BV-2^WT) and absent (RGS10^−/−) RGS10 expression, with or without interferon-γ (IFNγ)-stimulation. We identified genes whose expression were altered as a result of RGS10 loss, IFNγ stimulation, or both. As expected, RGS10 loss resulted in changes in processes such as GPCR signaling and synaptic signaling. However, this analysis also revealed that processes such as cell migration and adhesion were altered in RGS10^−/− cells, under both basal and IFNγ-stimulated conditions. Key cell adhesion genes such as Mmp9 and Thbs1 were downregulated in RGS10^−/− cells regardless of stimuli. Experimentally, loss of RGS10 increased BV-2 cell migration, and IFNγ stimulation led to a reduction in migration of both BV-2^WT and RGS10^−/− cells. The effect of RGS10 on migration could only partially be blocked by the Gα_i inhibitor Pertussis toxin (PTX), suggesting involvement of both G protein-dependent and -independent mechanisms. Altogether, these results suggest a novel role for RGS10 in reducing microglial migration through regulation of cell adhesion genes.

G蛋白信号传导10调节因子（RGS10）是一种GTPase激活蛋白，对Gαi具有选择性，已被提出在抑制小胶质细胞驱动的神经炎症中发挥作用。RGS10在小胶质细胞中的表达受到炎症刺激和衰老的抑制，RGS10的缺失与细胞因子表达增加和神经退行性变有关。相反，在啮齿动物模型中，RGS10过表达提供了抗炎症刺激的保护，但RGS10发挥这种保护作用的机制尚不清楚。为了了解RGS10在小胶质细胞中的神经保护功能，我们对小鼠小胶质细胞系BV-2进行了RNA-Seq分析，在有或没有干扰素-γ (IFNγ)刺激的情况下，RGS10表达完整（BV-2WT）和缺失（RGS10-/-）。我们确定了由于RGS10缺失、IFNγ刺激或两者兼而有之而改变表达的基因。正如预期的那样，RGS10的缺失导致了GPCR信号传导和突触信号传导等过程的改变。然而，该分析还显示，在基础和ifn γ刺激条件下，RGS10-/-细胞的细胞迁移和粘附等过程发生了改变。在RGS10-/-细胞中，无论受到何种刺激，关键细胞粘附基因Mmp9和Thbs1均下调。实验表明，RGS10的缺失增加了BV-2细胞的迁移，而IFNγ刺激导致BV-2WT和RGS10-/-细胞的迁移减少。RGS10对迁移的影响只能被Gαi抑制剂百日咳毒素（Pertussis toxin， PTX）部分阻断，提示其参与了G蛋白依赖性和非依赖性机制。总之，这些结果表明RGS10通过调节细胞粘附基因在减少小胶质细胞迁移中的新作用。

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引用次数: 0