Pub Date : 2026-01-25DOI: 10.1016/j.phymed.2026.157884
Yijun Tian , Changjie Ren , Hongru Wang , Wei Yang , Jianqing Ye , Banglan Cai , Kuo Yang , Yongbin Chi , Denghai Zhang , Xiuwu Pan , Xue Zhang , Denglong Wu
Background
: Celastrol, a bioactive triterpenoid from Tripterygium wilfordii, exhibits antitumor properties across multiple malignancies, while the molecular mechanisms linking it to metabolic regulation and ferroptosis in clear cell renal cell carcinoma (ccRCC) remain insufficiently defined.
Purpose
: This study aimed to identify the key molecular mediator through which celastrol induces ferroptosis in ccRCC and to evaluate its clinical and prognostic relevance.
Methods
: Transcriptomics, bioinformatics, and functional assays were combined with clinical validation in ccRCC tissues (n=278) and TCGA datasets. Mechanistic analyses included chromatin immunoprecipitation, luciferase reporter assays, cellular thermal shift, molecular docking, and ubiquitination assays, with ferroptosis evaluated in vitro and in xenograft models.
Results
: ACOX2 was identified as a celastrol-responsive lipid metabolic enzyme with reduced expression in ccRCC. ACOX2 showed prognostic value for overall survival prediction (AUC = 0.8019), with low expression corresponding to unfavorable outcomes in both univariable (HR = 0.169, CI 0.064–0.440) and multivariable analyses (HR = 0.186, CI 0.067–0.517). Functionally, ACOX2 enhanced lipid peroxidation and ferroptotic sensitivity while restraining malignant phenotypes. Celastrol stabilized the transcription factor TEF by inhibiting its ubiquitination and proteasomal degradation, thereby promoting transcriptional activation of ACOX2. ChIP–qPCR demonstrated increased TEF occupancy at the ACOX2 promoter, with TEF/Input rising from 4.72 ± 1.23 to 8.08 ± 0.94 and TEF/IgG increasing from 4.71 ± 1.24 to 7.94 ± 2.02, indicating a ∼1.70-fold enhancement. In vivo, celastrol suppressed tumor growth and induced ferroptosis in an ACOX2-dependent manner without systemic toxicity.
Conclusion
: A TEF–ACOX2 regulatory axis is defined through which celastrol induces ferroptosis in ccRCC, linking peroxisomal lipid metabolism to tumor suppression. These findings identify ACOX2 as both a mechanistic mediator and a prognostic biomarker and support celastrol as a phytochemical candidate for ccRCC therapy.
背景：雷公藤红素是一种来自雷公藤的生物活性三萜，在多种恶性肿瘤中表现出抗肿瘤特性，但其与透明细胞肾细胞癌（ccRCC）代谢调节和铁凋亡相关的分子机制尚不明确。目的：本研究旨在确定雷公藤红素诱导ccRCC患者铁下垂的关键分子介质，并评估其临床和预后相关性。方法：将转录组学、生物信息学和功能分析与临床验证相结合，对278例ccRCC组织和TCGA数据集进行研究。机制分析包括染色质免疫沉淀、荧光素酶报告分析、细胞热移、分子对接和泛素化分析，并在体外和异种移植模型中评估铁下垂。结果：ACOX2在ccRCC中被鉴定为celastrol反应性脂质代谢酶，表达降低。ACOX2在总生存预测中具有预后价值（AUC = 0.8019），在单变量分析（HR = 0.169, CI 0.064-0.440）和多变量分析（HR = 0.186, CI 0.067-0.517）中，低表达均对应不良结果。在功能上，ACOX2增强了脂质过氧化和铁致敏感性，同时抑制了恶性表型。Celastrol通过抑制转录因子TEF的泛素化和蛋白酶体降解来稳定TEF，从而促进ACOX2的转录激活。ChIP-qPCR显示，ACOX2启动子的TEF占用率增加，TEF/Input从4.72±1.23增加到8.08±0.94，TEF/IgG从4.71±1.24增加到7.94±2.02，增加了约1.70倍。在体内，雷公藤红素以acox2依赖的方式抑制肿瘤生长并诱导铁下垂，无全身毒性。结论：在ccRCC中存在TEF-ACOX2调节轴，通过该调节轴，celastrol诱导铁细胞凋亡，将过氧化物酶体脂质代谢与肿瘤抑制联系起来。这些发现确定ACOX2既是一种机制介质，也是一种预后生物标志物，并支持celastrol作为ccRCC治疗的植物化学候选物。
{"title":"Celastrol induces ferroptosis in ccRCC through TEF-Driven ACOX2 upregulation and metabolic reprogramming","authors":"Yijun Tian , Changjie Ren , Hongru Wang , Wei Yang , Jianqing Ye , Banglan Cai , Kuo Yang , Yongbin Chi , Denghai Zhang , Xiuwu Pan , Xue Zhang , Denglong Wu","doi":"10.1016/j.phymed.2026.157884","DOIUrl":"10.1016/j.phymed.2026.157884","url":null,"abstract":"<div><h3>Background</h3><div><strong>:</strong> Celastrol, a bioactive triterpenoid from Tripterygium wilfordii, exhibits antitumor properties across multiple malignancies, while the molecular mechanisms linking it to metabolic regulation and ferroptosis in clear cell renal cell carcinoma (ccRCC) remain insufficiently defined.</div></div><div><h3>Purpose</h3><div><strong>:</strong> This study aimed to identify the key molecular mediator through which celastrol induces ferroptosis in ccRCC and to evaluate its clinical and prognostic relevance.</div></div><div><h3>Methods</h3><div><strong>:</strong> Transcriptomics, bioinformatics, and functional assays were combined with clinical validation in ccRCC tissues (n=278) and TCGA datasets. Mechanistic analyses included chromatin immunoprecipitation, luciferase reporter assays, cellular thermal shift, molecular docking, and ubiquitination assays, with ferroptosis evaluated in vitro and in xenograft models.</div></div><div><h3>Results</h3><div><strong>:</strong> ACOX2 was identified as a celastrol-responsive lipid metabolic enzyme with reduced expression in ccRCC. ACOX2 showed prognostic value for overall survival prediction (AUC = 0.8019), with low expression corresponding to unfavorable outcomes in both univariable (HR = 0.169, CI 0.064–0.440) and multivariable analyses (HR = 0.186, CI 0.067–0.517). Functionally, ACOX2 enhanced lipid peroxidation and ferroptotic sensitivity while restraining malignant phenotypes. Celastrol stabilized the transcription factor TEF by inhibiting its ubiquitination and proteasomal degradation, thereby promoting transcriptional activation of ACOX2. ChIP–qPCR demonstrated increased TEF occupancy at the ACOX2 promoter, with TEF/Input rising from 4.72 ± 1.23 to 8.08 ± 0.94 and TEF/IgG increasing from 4.71 ± 1.24 to 7.94 ± 2.02, indicating a ∼1.70-fold enhancement. In vivo, celastrol suppressed tumor growth and induced ferroptosis in an ACOX2-dependent manner without systemic toxicity.</div></div><div><h3>Conclusion</h3><div><strong>:</strong> A TEF–ACOX2 regulatory axis is defined through which celastrol induces ferroptosis in ccRCC, linking peroxisomal lipid metabolism to tumor suppression. These findings identify ACOX2 as both a mechanistic mediator and a prognostic biomarker and support celastrol as a phytochemical candidate for ccRCC therapy.</div></div>","PeriodicalId":20212,"journal":{"name":"Phytomedicine","volume":"152 ","pages":"Article 157884"},"PeriodicalIF":8.3,"publicationDate":"2026-01-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146080566","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2026-01-25DOI: 10.1016/j.phymed.2026.157886
Yingying Xie , Haoming He , Yike Li , Qiang Chen , Sunjing Fu , Zhe Wang , Gaiyan Feng , Yanping Li , YanXiang Gao , Jingang Zheng
Background
Heart failure with preserved ejection fraction (HFpEF) constitutes over 50% of heart failure cases but lacks disease-modifying therapies. The pathophysiological role of eicosapentaenoic acid (EPA) in HFpEF remains undefined.
Methods
Integrated lipidomics was conducted across HFpEF discovery and validation cohorts. "Two-hit" murine HFpEF model combining high-fat diet (HFD) and 0.5 g/l-NAME was established to recapitulate human metabolic-inflammatory pathology. EPA’s efficacy was evaluated through prophylactic/therapeutic interventions (160/320 mg/kg/day, human-equivalent 2/4 g/day). Mechanistic studies integrated transcriptomics, molecular docking, triggering receptor expressed on myeloid cells 2 (TREM2) knockout, and siRNA silencing.
Results
Plasma EPA deficiency correlated with diastolic dysfunction severity and conferred incremental diagnostic value. High-dose EPA (4 g/day equivalent) prevented/reversed diastolic impairment and apoptosis in HFpEF mice. EPA rescued impaired efferocytosis through dual modulation of TREM2, concurrently enhancing functional transmembrane receptor expression while suppressing pathological ectodomain shedding. TREM2 ablation attenuated EPA-mediated benefits on diastolic function and efferocytosis.
Conclusion
Our work identifies plasma EPA depletion as a potential biomarker for risk stratification and delineates the EPA-TREM2-efferocytosis axis as a putative therapeutic mechanism for HFpEF, suggesting the potential of EPA as a theranostic candidate.
{"title":"Eicosapentaenoic acid attenuates heart failure with preserved ejection fraction via promoting TREM2-dependent efferocytosis","authors":"Yingying Xie , Haoming He , Yike Li , Qiang Chen , Sunjing Fu , Zhe Wang , Gaiyan Feng , Yanping Li , YanXiang Gao , Jingang Zheng","doi":"10.1016/j.phymed.2026.157886","DOIUrl":"10.1016/j.phymed.2026.157886","url":null,"abstract":"<div><h3>Background</h3><div>Heart failure with preserved ejection fraction (HFpEF) constitutes over 50% of heart failure cases but lacks disease-modifying therapies. The pathophysiological role of eicosapentaenoic acid (EPA) in HFpEF remains undefined.</div></div><div><h3>Methods</h3><div>Integrated lipidomics was conducted across HFpEF discovery and validation cohorts. \"Two-hit\" murine HFpEF model combining high-fat diet (HFD) and 0.5 g/l-NAME was established to recapitulate human metabolic-inflammatory pathology. EPA’s efficacy was evaluated through prophylactic/therapeutic interventions (160/320 mg/kg/day, human-equivalent 2/4 g/day). Mechanistic studies integrated transcriptomics, molecular docking, triggering receptor expressed on myeloid cells 2 (TREM2) knockout, and siRNA silencing.</div></div><div><h3>Results</h3><div>Plasma EPA deficiency correlated with diastolic dysfunction severity and conferred incremental diagnostic value. High-dose EPA (4 g/day equivalent) prevented/reversed diastolic impairment and apoptosis in HFpEF mice. EPA rescued impaired efferocytosis through dual modulation of TREM2, concurrently enhancing functional transmembrane receptor expression while suppressing pathological ectodomain shedding. TREM2 ablation attenuated EPA-mediated benefits on diastolic function and efferocytosis.</div></div><div><h3>Conclusion</h3><div>Our work identifies plasma EPA depletion as a potential biomarker for risk stratification and delineates the EPA-TREM2-efferocytosis axis as a putative therapeutic mechanism for HFpEF, suggesting the potential of EPA as a theranostic candidate.</div></div>","PeriodicalId":20212,"journal":{"name":"Phytomedicine","volume":"153 ","pages":"Article 157886"},"PeriodicalIF":8.3,"publicationDate":"2026-01-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146143257","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2026-01-24DOI: 10.1016/j.phymed.2026.157883
Liang Li , Pengfei Liu , Chang Zhang , Xinyi Yu , Jun Tao , Zhengzheng Zhou
<div><h3>Background</h3><div>Ge-Gen-Huang-Lian Decoction (GGHLD) is a traditional Chinese herbal formula with diverse pharmacological effects, primarily attributed to puerarin (PUE, from <em>Pueraria lobata</em>) and berberine (BER, from Coptidis rhizoma). However, the limited solubility and bioavailability of PUE and BER restrict their clinical efficacy.</div></div><div><h3>Purpose</h3><div>To address these limitations, a novel PUE-BER cocrystal was engineered to enhance solubility, dissolution rate, phase stability, and <em>in vivo</em> bioavailability, with structural mechanisms elucidated through combined experimental and computational approaches.</div></div><div><h3>Study design</h3><div>A multidisciplinary comparative study integrating crystal engineering, physicochemical characterization, <em>in vitro</em> dissolution/stability testing, <em>in vivo</em> pharmacokinetics in rats, and computational lattice analysis.</div></div><div><h3>Methods</h3><div>The PUE-BER cocrystal was synthesized <em>via</em> crystal engineering and characterized by single crystal X-ray diffraction (SCXRD). Phase stability was evaluated by powder X-ray diffraction (PXRD) in water and pH 1.2/6.8 buffers. Equilibrium solubility and intrinsic dissolution rate (IDR) were determined for PUE and BER. Pharmacokinetics were assessed in Sprague-Dawley rats (n = 8/group) after oral administration of the cocrystal, pure compounds, and simulated GGHLD; plasma levels were quantified by UPLC-MS/MS, and pharmacokinetic parameters (C<sub>max</sub>, <em>AUC<sub>0</sub><sub>–</sub><sub>24</sub>, T<sub>max</sub></em>) calculated <em>via</em> non-compartmental analysis. Computational modeling included lattice energy, packing efficiency, hydrogen bond density, and channel dimensions using SCXRD data.</div></div><div><h3>Results</h3><div>SCXRD revealed a low-density orthorhombic <em>P</em> 2<sub>1</sub> 2<sub>1</sub> 2<sub>1</sub> lattice (1.502 g/cm³) with 65 % packing efficiency, a dense hydrogen bond network (0.25 H-bonds/atom), and tubular solvent channels (7.655 Å interlayer spacing). <em>In vitro</em>, PUE solubility increased 2.0-fold in water, 1.5-fold in pH 1.2 buffer, and 1.3-fold in pH 6.8 buffer <em>versus</em> pure PUE, with significantly higher IDR; BER solubility was moderately reduced. PXRD confirmed phase stability across all media. <em>In vivo</em>, the cocrystal increased PUE <em>C<sub>max</sub></em> by 1.8-fold (104.1 ± 18.1 μg/L <em>vs.</em> 57.2 ± 7.2 μg/L), <em>AUC<sub>0</sub><sub>–</sub><sub>24</sub></em> by 3.0-fold (384.4 ± 20.5 μg/L.h <em>vs.</em> 127.6 ± 10.8 μg/L.h), and prolonged <em>T<sub>max</sub></em> (1.4 ± 0.1 h <em>vs.</em> 1.1 ± 0.1 h), with secondary peaks indicating enterohepatic recirculation. For BER, <em>C<sub>max</sub></em> increased 1.2-fold (10.2 ± 0.9 μg/L <em>vs.</em> 8.8 ± 1.6 μg/L) and <em>AUC<sub>0</sub><sub>–</sub><sub>24</sub></em> 1.4-fold (55.3 ± 3.7 μg/L.h <em>vs.</em> 40.2 ± 2.8 μg/L.h). Compared with simulated GGHLD, the cocrystal
葛根黄连汤（GGHLD）是一种具有多种药理作用的传统中药配方，其主要成分是葛根素（PUE，来自葛根）和小檗碱（BER，来自黄连）。然而，PUE和BER有限的溶解度和生物利用度限制了它们的临床疗效。为了解决这些局限性，设计了一种新型PUE-BER共晶，以提高其溶解度、溶解速度、相稳定性和体内生物利用度，并通过实验和计算相结合的方法阐明了其结构机制。研究设计一项多学科比较研究，包括晶体工程、理化表征、体外溶出/稳定性测试、大鼠体内药代动力学和计算晶格分析。方法采用晶体工程方法合成PUE-BER共晶，并用单晶x射线衍射（SCXRD）对其进行表征。在pH为1.2/6.8的缓冲液中，采用粉末x射线衍射（PXRD）评价相稳定性。测定了PUE和BER的平衡溶解度和固有溶出率（IDR）。口服共晶、纯化合物和模拟GGHLD后，在Sprague-Dawley大鼠（n = 8/组）中评估药代动力学；采用超高效液相色谱-质谱联用（UPLC-MS/MS）法测定血药浓度，通过非室室分析计算药代动力学参数（Cmax、AUC0-24、Tmax）。计算模型包括晶格能量、填充效率、氢键密度和利用SCXRD数据的通道尺寸。结果scxrd结果显示，p21 21 21晶格为低密度正交晶格（1.502 g/cm³），填充效率为65%，具有致密的氢键网络（0.25个氢键/原子）和管状溶剂通道（7.655 Å层间距）。在体外，PUE在水中的溶解度比纯PUE提高2.0倍，在pH 1.2缓冲液中提高1.5倍，在pH 6.8缓冲液中提高1.3倍，IDR显著提高；BER的溶解度适度降低。PXRD证实了所有介质的相稳定性。体内PUE Cmax提高1.8倍（104.1±18.1 μg/L vs. 57.2±7.2 μg/L）， AUC0-24提高3.0倍（384.4±20.5 μg/L h vs. 127.6±10.8 μg/L）， Tmax延长（1.4±0.1 h vs. 1.1±0.1 h），次峰表明肠肝再循环。对于BER， Cmax增加1.2倍（10.2±0.9 μg/L vs 8.8±1.6 μg/L）， AUC0-24增加1.4倍（55.3±3.7 μg/L h vs 40.2±2.8 μg/L）。与模拟GGHLD相比，该共晶体显著提高了PUE的生物利用度，并适度提高了BER。计算模型预测，由于溶剂可达通道促进溶解能垒的降低，IDR增强。结论PUE-BER共晶可显著提高PUE的溶解度、IDR和口服生物利用度，同时保持稳定性并适度增加BER暴露。计算洞察力将这些增强与优化的晶格结构和溶剂通道联系起来，建立了共晶作为GGHLD衍生疗法的有前途的配方策略。
{"title":"Herbal-derived puerarin-berberine cocrystal: Computational insights into mechanisms driving simultaneous enhanced solubility and bioavailability","authors":"Liang Li , Pengfei Liu , Chang Zhang , Xinyi Yu , Jun Tao , Zhengzheng Zhou","doi":"10.1016/j.phymed.2026.157883","DOIUrl":"10.1016/j.phymed.2026.157883","url":null,"abstract":"<div><h3>Background</h3><div>Ge-Gen-Huang-Lian Decoction (GGHLD) is a traditional Chinese herbal formula with diverse pharmacological effects, primarily attributed to puerarin (PUE, from <em>Pueraria lobata</em>) and berberine (BER, from Coptidis rhizoma). However, the limited solubility and bioavailability of PUE and BER restrict their clinical efficacy.</div></div><div><h3>Purpose</h3><div>To address these limitations, a novel PUE-BER cocrystal was engineered to enhance solubility, dissolution rate, phase stability, and <em>in vivo</em> bioavailability, with structural mechanisms elucidated through combined experimental and computational approaches.</div></div><div><h3>Study design</h3><div>A multidisciplinary comparative study integrating crystal engineering, physicochemical characterization, <em>in vitro</em> dissolution/stability testing, <em>in vivo</em> pharmacokinetics in rats, and computational lattice analysis.</div></div><div><h3>Methods</h3><div>The PUE-BER cocrystal was synthesized <em>via</em> crystal engineering and characterized by single crystal X-ray diffraction (SCXRD). Phase stability was evaluated by powder X-ray diffraction (PXRD) in water and pH 1.2/6.8 buffers. Equilibrium solubility and intrinsic dissolution rate (IDR) were determined for PUE and BER. Pharmacokinetics were assessed in Sprague-Dawley rats (n = 8/group) after oral administration of the cocrystal, pure compounds, and simulated GGHLD; plasma levels were quantified by UPLC-MS/MS, and pharmacokinetic parameters (C<sub>max</sub>, <em>AUC<sub>0</sub><sub>–</sub><sub>24</sub>, T<sub>max</sub></em>) calculated <em>via</em> non-compartmental analysis. Computational modeling included lattice energy, packing efficiency, hydrogen bond density, and channel dimensions using SCXRD data.</div></div><div><h3>Results</h3><div>SCXRD revealed a low-density orthorhombic <em>P</em> 2<sub>1</sub> 2<sub>1</sub> 2<sub>1</sub> lattice (1.502 g/cm³) with 65 % packing efficiency, a dense hydrogen bond network (0.25 H-bonds/atom), and tubular solvent channels (7.655 Å interlayer spacing). <em>In vitro</em>, PUE solubility increased 2.0-fold in water, 1.5-fold in pH 1.2 buffer, and 1.3-fold in pH 6.8 buffer <em>versus</em> pure PUE, with significantly higher IDR; BER solubility was moderately reduced. PXRD confirmed phase stability across all media. <em>In vivo</em>, the cocrystal increased PUE <em>C<sub>max</sub></em> by 1.8-fold (104.1 ± 18.1 μg/L <em>vs.</em> 57.2 ± 7.2 μg/L), <em>AUC<sub>0</sub><sub>–</sub><sub>24</sub></em> by 3.0-fold (384.4 ± 20.5 μg/L.h <em>vs.</em> 127.6 ± 10.8 μg/L.h), and prolonged <em>T<sub>max</sub></em> (1.4 ± 0.1 h <em>vs.</em> 1.1 ± 0.1 h), with secondary peaks indicating enterohepatic recirculation. For BER, <em>C<sub>max</sub></em> increased 1.2-fold (10.2 ± 0.9 μg/L <em>vs.</em> 8.8 ± 1.6 μg/L) and <em>AUC<sub>0</sub><sub>–</sub><sub>24</sub></em> 1.4-fold (55.3 ± 3.7 μg/L.h <em>vs.</em> 40.2 ± 2.8 μg/L.h). Compared with simulated GGHLD, the cocrystal","PeriodicalId":20212,"journal":{"name":"Phytomedicine","volume":"152 ","pages":"Article 157883"},"PeriodicalIF":8.3,"publicationDate":"2026-01-24","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146080058","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2026-01-24DOI: 10.1016/j.phymed.2026.157887
Chang ZHOU , Ye TANG , Fuchang LU , Cheng-hao DU , Wei-liang ZHU , Gui-xian CHEN , Hui XIA , Min ZHAO , Ze-quan ZHENG , Yuan-qi ZHAO
Background
Stroke imposes a substantial global health burden. Microglial pyroptosis promotes acute cerebral ischemia. Gasdermin D (GSDMD), the principal executor of pyroptosis, has been implicated in a newly characterized form of proinflammatory cell death known as extracellular trap death (ETosis). Thus, targeting GSDMD may represent a promising therapeutic strategy to mitigate ischemic stroke. Xing-nao-sheng-jiang powder (XNSJP) was demonstrated to inhibit GSDMD-mediated pyroptosis in cerebral ischemia-reperfusion injury (CIRI). Based on these findings, we hypothesize that XNSJP may inhibit pyroptosis-related ETosis by targeting the GSDMD, thereby alleviating neuroinflammation in CIRI.
Methods
We prepared the MCAO model in SD rats and evaluated the effects of XNSJP on anti-CIRI. The effects of XNSJP on microglia and neutrophil pyroptosis and ETosis were detected. The composition of XNSJP was identified using UPLC and MS/MS methods.
Results
UPLC identified the characteristic peaks of XNSJP’s active ingredients. MS/MS and network pharmacology suggested that XNSJP has regulatory effects on atherosclerosis, coagulation, and inflammation. The XNSJP reduced the area of infarction. XNSJP could inhibit the caspase-1/11/GSDMD-mediated pyroptosis. Furthermore, XNSJP impeded microglial pyroptosis-related ETosis by GSDMD, as evidenced by the suppression of MPO, PAD4, and CitH3. We also confirmed that 24 h after CIRI, the ETosis marker CitH3 mainly co-localized with microglia.
Conclusions
We confirmed that microglia are crucial contributors to ETosis in CIRI. Furthermore, we demonstrated that XNSJP inhibited microglial pyroptosis and ETosis against CIRI, which is closely related to the inhibition of GSDMD. Mechanistically, XNSJP inhibits caspase-1/11/GSDMD-mediated microglial pyroptosis and MPO/PAD4/CitH3-mediated microglial ETosis.
背景：脑卒中造成了巨大的全球健康负担。小胶质细胞焦亡促进急性脑缺血。Gasdermin D （GSDMD）是焦亡的主要执行者，与一种新特征的促炎细胞死亡形式（称为细胞外陷阱死亡（ETosis））有关。因此，靶向GSDMD可能是缓解缺血性卒中的一种有前景的治疗策略。醒脑生姜散（XNSJP）对脑缺血再灌注损伤（CIRI）有明显的抑制作用。基于这些发现，我们假设XNSJP可能通过靶向GSDMD来抑制焦热相关的ETosis，从而减轻CIRI的神经炎症。方法：制备SD大鼠MCAO模型，评价XNSJP抗ciri的作用。观察XNSJP对小胶质细胞和中性粒细胞焦亡和ETosis的影响。采用超高效液相色谱法和质谱法对XNSJP的成分进行了鉴定。结果：超高效液相色谱法鉴定出了XNSJP有效成分的特征峰。MS/MS和网络药理学提示XNSJP对动脉粥样硬化、凝血和炎症具有调节作用。XNSJP减少了梗死面积。XNSJP能抑制caspase-1/11/ gsdmd介导的焦亡。此外，XNSJP可以抑制MPO、PAD4和CitH3，从而抑制GSDMD引起的小胶质细胞热降解相关的ETosis。我们还证实，CIRI后24小时，ETosis标记物CitH3主要与小胶质细胞共定位。结论：我们证实小胶质细胞是CIRI中ETosis的关键贡献者。此外，我们证明XNSJP抑制CIRI的小胶质细胞焦亡和ETosis，这与抑制GSDMD密切相关。机制上，XNSJP抑制caspase-1/11/ gsdmd介导的小胶质细胞焦亡和MPO/PAD4/ cith3介导的小胶质细胞凋亡。
{"title":"Xing-nao-sheng-jiang powder alleviates ischemic stroke in rats by inhibiting pyroptosis-related microglial ETosis: An emerging perspective on microglial ETosis","authors":"Chang ZHOU , Ye TANG , Fuchang LU , Cheng-hao DU , Wei-liang ZHU , Gui-xian CHEN , Hui XIA , Min ZHAO , Ze-quan ZHENG , Yuan-qi ZHAO","doi":"10.1016/j.phymed.2026.157887","DOIUrl":"10.1016/j.phymed.2026.157887","url":null,"abstract":"<div><h3>Background</h3><div>Stroke imposes a substantial global health burden. Microglial pyroptosis promotes acute cerebral ischemia. Gasdermin D (GSDMD), the principal executor of pyroptosis, has been implicated in a newly characterized form of proinflammatory cell death known as extracellular trap death (ETosis). Thus, targeting GSDMD may represent a promising therapeutic strategy to mitigate ischemic stroke. Xing-nao-sheng-jiang powder (XNSJP) was demonstrated to inhibit GSDMD-mediated pyroptosis in cerebral ischemia-reperfusion injury (CIRI). Based on these findings, we hypothesize that XNSJP may inhibit pyroptosis-related ETosis by targeting the GSDMD, thereby alleviating neuroinflammation in CIRI.</div></div><div><h3>Methods</h3><div>We prepared the MCAO model in SD rats and evaluated the effects of XNSJP on anti-CIRI. The effects of XNSJP on microglia and neutrophil pyroptosis and ETosis were detected. The composition of XNSJP was identified using UPLC and MS/MS methods.</div></div><div><h3>Results</h3><div>UPLC identified the characteristic peaks of XNSJP’s active ingredients. MS/MS and network pharmacology suggested that XNSJP has regulatory effects on atherosclerosis, coagulation, and inflammation. The XNSJP reduced the area of infarction. XNSJP could inhibit the caspase-1/11/GSDMD-mediated pyroptosis. Furthermore, XNSJP impeded microglial pyroptosis-related ETosis by GSDMD, as evidenced by the suppression of MPO, PAD4, and CitH3. We also confirmed that 24 h after CIRI, the ETosis marker CitH3 mainly co-localized with microglia.</div></div><div><h3>Conclusions</h3><div>We confirmed that microglia are crucial contributors to ETosis in CIRI. Furthermore, we demonstrated that XNSJP inhibited microglial pyroptosis and ETosis against CIRI, which is closely related to the inhibition of GSDMD. Mechanistically, XNSJP inhibits caspase-1/11/GSDMD-mediated microglial pyroptosis and MPO/PAD4/CitH3-mediated microglial ETosis.</div></div>","PeriodicalId":20212,"journal":{"name":"Phytomedicine","volume":"153 ","pages":"Article 157887"},"PeriodicalIF":8.3,"publicationDate":"2026-01-24","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146158144","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2026-01-24DOI: 10.1016/j.phymed.2026.157885
Zhixuan Deng , Woding Deng , Ning Tang , Chunmei Ban , Yupeng Wu , Mengqi Gong , Peng Wu , Xuewei Bao , Hao Zhou , Yikao Liu , Xin Wu , Qiangqiang Zhao
Background
Acute myeloid leukemia (AML) is a heterogeneous hematological malignancy characterized by genetic diversity. These features of AML contribute to immune evasion and therapeutic resistance. Relapse after chemotherapy remains a major clinical challenge in the management of AML. The bone marrow microenvironment is believed to play a critical role in resistance; however, the underlying mechanisms are not fully understood.
Methods
We combined spatial single-cell analyzes, ligand–receptor network inference, and Treg pseudotime modeling with functional genetics and pharmacology. A full experimental cascade (reverse transcription [RT]-quantitative polymerase chain reaction [qPCR], western blotting, immunofluorescence, and flow cytometry/function) and in vivo xenografts were used to validate these mechanisms. Structure-guided modeling supported the direct recognition of TRIM44 by the natural alkaloid sinomenine, complemented by cellular evidence of its on-target activity.
Results
Single-cell profiling revealed increased immune heterogeneity and enrichment of regulatory T cells (Tregs) in relapsed AML. TRIM44 was overexpressed during relapse and is associated with poor outcomes. Spatial mapping and trajectory analysis revealed a TRIM44-driven transition from Tregnaïve to Tregeffector, and tumor–Treg co-culture showed that TRIM44 knockdown in AML cells dampens Treg activation/effectorization. Mechanistically, TRIM44 coordinated a dual resistance program—apoptosis escape and epithelial–mesenchymal transition (EMT)-like features in leukemic blasts—converging with enhanced tumor→Treg communication. Sinomenine disrupted this TRIM44-dependent axis, suppressed Treg activity, and re-sensitized resistant AML cells to cytarabine, yielding synergistic anti-leukemic effects in vitro and efficacy in xenograft models.
Conclusions
This study coupled spatial single-cell multiomics with functional genetics and pharmacology to decode an immune-metabolic driver of AML relapse, providing experimental evidence that pharmacological TRIM44 blockade overturns Treg-mediated immune evasion and chemoresistance. We nominate sinomenine as a tractable TRIM44 inhibitor and propose the TRIM44–Treg axis as an actionable therapeutic entry point for precision combinations with cytarabine in relapsed AML.
背景：急性髓系白血病（AML）是一种以遗传多样性为特征的异质性血液系统恶性肿瘤。AML的这些特征有助于免疫逃避和治疗抵抗。化疗后复发仍然是AML治疗的主要临床挑战。骨髓微环境被认为在耐药性中起关键作用；然而，其潜在机制尚未完全了解。方法：将空间单细胞分析、配体-受体网络推断、Treg伪时间模型与功能遗传学和药理学相结合。完整的实验级联（逆转录[RT]-定量聚合酶链反应[qPCR]， western blotting，免疫荧光和流式细胞术/功能）和体内异种移植物用于验证这些机制。结构引导模型支持天然生物碱青藤碱对TRIM44的直接识别，并辅以其靶向活性的细胞证据。结果：单细胞分析显示，复发性AML患者的免疫异质性增加，调节性T细胞（Tregs）富集。TRIM44在复发期间过表达，并与不良预后相关。空间定位和轨迹分析揭示了TRIM44驱动的从Tregnaïve到tregeeffector的转变，肿瘤-Treg共培养表明，AML细胞中TRIM44敲低抑制Treg的激活/效应。在机制上，TRIM44协调了白血病细胞的双重耐药程序-细胞凋亡逃逸和上皮-间质转化（EMT）样特征-与增强的肿瘤→Treg通讯趋同。青藤碱破坏了这条依赖trim44的轴，抑制Treg活性，并使耐药的AML细胞对阿糖胞苷重新敏感，在体外和异种移植模型中产生协同抗白血病作用。结论：本研究将空间单细胞多组学与功能遗传学和药理学结合，解码了AML复发的免疫代谢驱动因素，为药物阻断TRIM44推翻treg介导的免疫逃避和化疗耐药提供了实验证据。我们提名青藤碱作为一种易于处理的TRIM44抑制剂，并提出TRIM44- treg轴作为复发性AML精确联合阿糖胞苷的可行治疗切入点。
{"title":"Spatial-single cell multiomics reveals TRIM44-driven Treg differentiation and drug resistance in AML: Therapeutic reversal by Sinomenine","authors":"Zhixuan Deng , Woding Deng , Ning Tang , Chunmei Ban , Yupeng Wu , Mengqi Gong , Peng Wu , Xuewei Bao , Hao Zhou , Yikao Liu , Xin Wu , Qiangqiang Zhao","doi":"10.1016/j.phymed.2026.157885","DOIUrl":"10.1016/j.phymed.2026.157885","url":null,"abstract":"<div><h3>Background</h3><div>Acute myeloid leukemia (AML) is a heterogeneous hematological malignancy characterized by genetic diversity. These features of AML contribute to immune evasion and therapeutic resistance. Relapse after chemotherapy remains a major clinical challenge in the management of AML. The bone marrow microenvironment is believed to play a critical role in resistance; however, the underlying mechanisms are not fully understood.</div></div><div><h3>Methods</h3><div>We combined spatial single-cell analyzes, ligand–receptor network inference, and Treg pseudotime modeling with functional genetics and pharmacology. A full experimental cascade (reverse transcription [RT]-quantitative polymerase chain reaction [qPCR], western blotting, immunofluorescence, and flow cytometry/function) and in vivo xenografts were used to validate these mechanisms. Structure-guided modeling supported the direct recognition of TRIM44 by the natural alkaloid sinomenine, complemented by cellular evidence of its on-target activity.</div></div><div><h3>Results</h3><div>Single-cell profiling revealed increased immune heterogeneity and enrichment of regulatory T cells (Tregs) in relapsed AML. TRIM44 was overexpressed during relapse and is associated with poor outcomes. Spatial mapping and trajectory analysis revealed a TRIM44-driven transition from Treg<sub>naïve</sub> to Treg<sub>effector</sub>, and tumor–Treg co-culture showed that TRIM44 knockdown in AML cells dampens Treg activation/effectorization. Mechanistically, TRIM44 coordinated a dual resistance program—apoptosis escape and epithelial–mesenchymal transition (EMT)-like features in leukemic blasts—converging with enhanced tumor→Treg communication. Sinomenine disrupted this TRIM44-dependent axis, suppressed Treg activity, and re-sensitized resistant AML cells to cytarabine, yielding synergistic anti-leukemic effects in vitro and efficacy in xenograft models.</div></div><div><h3>Conclusions</h3><div>This study coupled spatial single-cell multiomics with functional genetics and pharmacology to decode an immune-metabolic driver of AML relapse, providing experimental evidence that pharmacological TRIM44 blockade overturns Treg-mediated immune evasion and chemoresistance. We nominate sinomenine as a tractable TRIM44 inhibitor and propose the TRIM44–Treg axis as an actionable therapeutic entry point for precision combinations with cytarabine in relapsed AML.</div></div>","PeriodicalId":20212,"journal":{"name":"Phytomedicine","volume":"153 ","pages":"Article 157885"},"PeriodicalIF":8.3,"publicationDate":"2026-01-24","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146137879","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
<div><h3>Background</h3><div>Qingfei Jiedu Huatan Formula (QJHF) has demonstrated efficacy in reducing the clinical stabilization time for pneumonia patients, decreasing ICU durations, and enhancing patients' quality of life.</div></div><div><h3>Objective</h3><div>Protecting the alveolar epithelial barrier is a proven strategy for improving pneumonia outcomes. This study aims to investigate the protective effects and underlying mechanisms of QJHF on the alveolar epithelial barrier in pneumonia.</div></div><div><h3>Methods</h3><div>A mouse model of pneumonia and acute lung injury was established via tracheal instillation of Klebsiella pneumoniae and LPS to evaluate the therapeutic effects of QJHF. Additionally, a TNF-α-induced alveolar epithelial cell barrier injury model was employed to explore the molecular mechanisms underlying QJHF's protective effects. Transcriptomics combined with network analysis was utilized to identify the active components and mechanisms of action for QJHF. Molecular docking and molecular dynamics analyses elucidated the binding interactions between Ginsenoside Rb1 (GRb1) and target proteins, while the application of Nrf2 inhibitor and siRNA clarified the role of Nrf2 activation in GRb1-mediated alveolar barrier protection.</div></div><div><h3>Results</h3><div>QJHF significantly ameliorates Klebsiella pneumoniae-induced pneumonia in mice by improving lung tissue damage, reducing inflammatory cell infiltration, and decreasing levels of pro-inflammatory factors such as TNF-α, IL-1β, and IL-6. Additionally, QJHF upregulates the expression of cell junction proteins ZO-1 and OCLN, thereby protecting the alveolar epithelial barrier. Similarly, QJHF mitigates LPS-induced lung damage, inflammation, and alveolar epithelial barrier dysfunction. Through integrating serum component profiling, network pharmacology, and lung transcriptomics revealed that 12 constituents in QJHF may interact with Nrf2 to modulate oxidative stress and apoptosis signaling pathways. Both in vitro and in vivo studies demonstrate that GRb1, one of the 12 components, significantly protects against TNF-α-induced alveolar epithelial cell barrier dysfunction and LPS-induced lung inflammation and alveolar barrier damage in mice. Furthermore, GRb1 can inhibit TNF-α-induced ROS levels and apoptosis in alveolar epithelial cells. Molecular dynamics stimulation and luciferase assays demonstrate that GRb1 can bind to and activate Nrf2, promoting its interaction with antioxidant response elements. Additionally, rotenone can counteract the protective effect against apoptosis in the alveolar epithelial cell barrier by increasing cellular ROS levels. Similarly, siRNA can significantly impair GRb1′s protective effects against TNF-α-induced damage to the alveolar epithelial cell barrier. Nrf2 inhibitors notably attenuated GRb1′s amelioration of LPS-induced lung inflammation and alveolar barrier damage in mice.</div></div><div><h3>Conclusion</h3><div>This study confirms that
{"title":"Qingfei Jiedu Huatan formula protects against pneumonia by activating Nrf2 to suppress alveolar epithelial apoptosis","authors":"Yuhang Jiang , Qin Zhang , Yumeng Cheng , Haiyang Cao , Xiaoxiang Xing , Baixi Shan , Peng Zhao , Jiansheng Li","doi":"10.1016/j.phymed.2026.157879","DOIUrl":"10.1016/j.phymed.2026.157879","url":null,"abstract":"<div><h3>Background</h3><div>Qingfei Jiedu Huatan Formula (QJHF) has demonstrated efficacy in reducing the clinical stabilization time for pneumonia patients, decreasing ICU durations, and enhancing patients' quality of life.</div></div><div><h3>Objective</h3><div>Protecting the alveolar epithelial barrier is a proven strategy for improving pneumonia outcomes. This study aims to investigate the protective effects and underlying mechanisms of QJHF on the alveolar epithelial barrier in pneumonia.</div></div><div><h3>Methods</h3><div>A mouse model of pneumonia and acute lung injury was established via tracheal instillation of Klebsiella pneumoniae and LPS to evaluate the therapeutic effects of QJHF. Additionally, a TNF-α-induced alveolar epithelial cell barrier injury model was employed to explore the molecular mechanisms underlying QJHF's protective effects. Transcriptomics combined with network analysis was utilized to identify the active components and mechanisms of action for QJHF. Molecular docking and molecular dynamics analyses elucidated the binding interactions between Ginsenoside Rb1 (GRb1) and target proteins, while the application of Nrf2 inhibitor and siRNA clarified the role of Nrf2 activation in GRb1-mediated alveolar barrier protection.</div></div><div><h3>Results</h3><div>QJHF significantly ameliorates Klebsiella pneumoniae-induced pneumonia in mice by improving lung tissue damage, reducing inflammatory cell infiltration, and decreasing levels of pro-inflammatory factors such as TNF-α, IL-1β, and IL-6. Additionally, QJHF upregulates the expression of cell junction proteins ZO-1 and OCLN, thereby protecting the alveolar epithelial barrier. Similarly, QJHF mitigates LPS-induced lung damage, inflammation, and alveolar epithelial barrier dysfunction. Through integrating serum component profiling, network pharmacology, and lung transcriptomics revealed that 12 constituents in QJHF may interact with Nrf2 to modulate oxidative stress and apoptosis signaling pathways. Both in vitro and in vivo studies demonstrate that GRb1, one of the 12 components, significantly protects against TNF-α-induced alveolar epithelial cell barrier dysfunction and LPS-induced lung inflammation and alveolar barrier damage in mice. Furthermore, GRb1 can inhibit TNF-α-induced ROS levels and apoptosis in alveolar epithelial cells. Molecular dynamics stimulation and luciferase assays demonstrate that GRb1 can bind to and activate Nrf2, promoting its interaction with antioxidant response elements. Additionally, rotenone can counteract the protective effect against apoptosis in the alveolar epithelial cell barrier by increasing cellular ROS levels. Similarly, siRNA can significantly impair GRb1′s protective effects against TNF-α-induced damage to the alveolar epithelial cell barrier. Nrf2 inhibitors notably attenuated GRb1′s amelioration of LPS-induced lung inflammation and alveolar barrier damage in mice.</div></div><div><h3>Conclusion</h3><div>This study confirms that","PeriodicalId":20212,"journal":{"name":"Phytomedicine","volume":"152 ","pages":"Article 157879"},"PeriodicalIF":8.3,"publicationDate":"2026-01-24","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146113969","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2026-01-24DOI: 10.1016/j.phymed.2026.157888
Qing Yin , Zhengqin Xiong , Xinya Wang , Tong-You Wade Wei , Jiayue Chao , Wu Jiang , Ying Chen , Ruijun Dong , Hu Zhang , Yi Yang , Kaizheng Gong , Hui Shen
Background
Pulmonary arterial hypertension (PAH) is characterized by vascular remodeling and right ventricular failure. Paeonol, a bioactive compound extracted from the root bark of Paeonia suffruticosa, exhibits anti-inflammatory, antioxidant, and anticancer effects. However, its efficacy on pulmonary vasculature in the context of PAH remains unknown.
Purpose
This study aims to evaluate the protective effects of paeonol in murine pulmonary hypertension (PH) model and identify its underlying molecular mechanism.
Methods
A murine PH model was used to assess the effect of paeonol on hemodynamics, echocardiography and pathology. In vitro, pulmonary arterial smooth muscle cells (PASMCs) proliferation and apoptosis were evaluated. RNA sequencing and network pharmacology were analyzed for pathway changes. Molecular docking and surface plasmon resonance (SPR) were employed to identify paeonol targets, which were further validated by Western blotting and immunofluorescence.
Results
Paeonol administration significantly reduced right ventricular systolic pressure (RVSP), right ventricular hypertrophy, and pulmonary vascular remodeling in PH mice. Echocardiography further demonstrated that paeonol preserved right ventricular structural and functional integrity. In vitro, paeonol inhibited PASMCs proliferation and promoted apoptosis. RNA sequencing and network pharmacology revealed that the drug effect was on the bone morphogenetic protein (BMP)/transforming growth factor-β (TGF-β) pathway. SPR analysis confirmed a high-affinity direct binding between paeonol and BRCA1/BRCA2-containing complex subunit 3 (BRCC3). Paeonol binding promotes the stability of BMP receptors and restores the BMP/TGF-β signaling balance.
Conclusion
Paeonol attenuates PH by rebalancing the BMP/TGF-β signaling through interacting with BRCC3 activation, thereby inhibiting vascular remodeling. These findings suggest that paeonol is a promising therapeutic candidate for PAH.
{"title":"Paeonol alleviates pulmonary arterial hypertension by activation of BRCC3","authors":"Qing Yin , Zhengqin Xiong , Xinya Wang , Tong-You Wade Wei , Jiayue Chao , Wu Jiang , Ying Chen , Ruijun Dong , Hu Zhang , Yi Yang , Kaizheng Gong , Hui Shen","doi":"10.1016/j.phymed.2026.157888","DOIUrl":"10.1016/j.phymed.2026.157888","url":null,"abstract":"<div><h3>Background</h3><div>Pulmonary arterial hypertension (PAH) is characterized by vascular remodeling and right ventricular failure. Paeonol, a bioactive compound extracted from the root bark of <em>Paeonia suffruticosa</em>, exhibits anti-inflammatory, antioxidant, and anticancer effects. However, its efficacy on pulmonary vasculature in the context of PAH remains unknown.</div></div><div><h3>Purpose</h3><div>This study aims to evaluate the protective effects of paeonol in murine pulmonary hypertension (PH) model and identify its underlying molecular mechanism.</div></div><div><h3>Methods</h3><div>A murine PH model was used to assess the effect of paeonol on hemodynamics, echocardiography and pathology. <em>In vitro,</em> pulmonary arterial smooth muscle cells (PASMCs) proliferation and apoptosis were evaluated. RNA sequencing and network pharmacology were analyzed for pathway changes. Molecular docking and surface plasmon resonance (SPR) were employed to identify paeonol targets, which were further validated by Western blotting and immunofluorescence.</div></div><div><h3>Results</h3><div>Paeonol administration significantly reduced right ventricular systolic pressure (RVSP), right ventricular hypertrophy, and pulmonary vascular remodeling in PH mice. Echocardiography further demonstrated that paeonol preserved right ventricular structural and functional integrity. <em>In vitro</em>, paeonol inhibited PASMCs proliferation and promoted apoptosis. RNA sequencing and network pharmacology revealed that the drug effect was on the bone morphogenetic protein (BMP)/transforming growth factor-β (TGF-β) pathway. SPR analysis confirmed a high-affinity direct binding between paeonol and BRCA1/BRCA2-containing complex subunit 3 (BRCC3). Paeonol binding promotes the stability of BMP receptors and restores the BMP/TGF-β signaling balance.</div></div><div><h3>Conclusion</h3><div>Paeonol attenuates PH by rebalancing the BMP/TGF-β signaling through interacting with BRCC3 activation, thereby inhibiting vascular remodeling. These findings suggest that paeonol is a promising therapeutic candidate for PAH.</div></div>","PeriodicalId":20212,"journal":{"name":"Phytomedicine","volume":"152 ","pages":"Article 157888"},"PeriodicalIF":8.3,"publicationDate":"2026-01-24","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146119613","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Acinetobacter baumannii is a Gram-negative bacterial pathogen responsible for severe nosocomial infections. Carbapenems, including meropenem and imipenem, are β-lactam antibiotics commonly used as a last-resort treatment for infections caused by A. baumannii. While it is recognized that green tea polyphenols can enhance the efficacy of carbapenems against A. baumannii, the underlying mechanisms of this synergy remain undefined.
Purpose
This study aims to investigate the synergistic actions of green tea aqueous extract and its key bioactive component, epigallocatechin gallate (EGCG), in combination with meropenem against A. baumannii BAA-1605 and to elucidate the associated mechanisms.
Methods
The synergism with meropenem was assessed using antibiotic susceptibility testing and fractional inhibitory concentration index (FICI) analysis. The mechanism study included β-lactamase activity, Hoechst dye accumulation, and DiOC₂(3) membrane potential measurements. The effect of meropenem and/or EGCG on purified penicillin-binding protein 2 (PBP2) was also evaluated by nuclear magnetic resonance (NMR) spectroscopy and scanning electron microscopy (SEM). Transcriptomic sequencing and bioinformatic analyses were performed to elucidate global gene expression changes of A. baumannii BAA-1605 following exposure to meropenem and/or EGCG.
Results
Green tea aqueous extract synergized with meropenem against A. baumannii BAA-1605 by lowering the minimum inhibitory concentration (MIC) by 4-fold (128 to 32 μg/mL). EGCG was determined as the primary synergistic component in the extract (FICI = 0.375). The mechanism study showed that EGCG does not influence β-lactamase production or meropenem-mediated PBP2 inhibition but contributes to overcoming resistance by inhibiting efflux pump activity. Furthermore, transcriptomic analysis revealed that EGCG may suppress the AdeABC efflux system and enhance meropenem influx via a YiaD-dependent porin pathway.
Conclusion
This study presents a dual role of efflux inhibition and influx facilitation by EGCG in restoring meropenem efficacy, supporting its potential as a promising adjuvant to restore β-lactam potency against A. baumannii or other Gram-negative bacteria harbouring homologs of AdeABC and YiaD proteins.
{"title":"Restoration of meropenem efficacy against multidrug-resistant Acinetobacter baumannii by green tea polyphenol EGCG: Dual targeting of efflux and porin pathways","authors":"Zijian Liang , Xiaoxu Chen , Ivanhoe K.H. Leung , Pangzhen Zhang","doi":"10.1016/j.phymed.2026.157871","DOIUrl":"10.1016/j.phymed.2026.157871","url":null,"abstract":"<div><h3>Background</h3><div><em>Acinetobacter baumannii</em> is a Gram-negative bacterial pathogen responsible for severe nosocomial infections. Carbapenems, including meropenem and imipenem, are β-lactam antibiotics commonly used as a last-resort treatment for infections caused by <em>A. baumannii</em>. While it is recognized that green tea polyphenols can enhance the efficacy of carbapenems against <em>A. baumannii</em>, the underlying mechanisms of this synergy remain undefined.</div></div><div><h3>Purpose</h3><div>This study aims to investigate the synergistic actions of green tea aqueous extract and its key bioactive component, epigallocatechin gallate (EGCG), in combination with meropenem against <em>A. baumannii</em> BAA-1605 and to elucidate the associated mechanisms.</div></div><div><h3>Methods</h3><div>The synergism with meropenem was assessed using antibiotic susceptibility testing and fractional inhibitory concentration index (FICI) analysis. The mechanism study included β-lactamase activity, Hoechst dye accumulation, and DiOC₂(3) membrane potential measurements. The effect of meropenem and/or EGCG on purified penicillin-binding protein 2 (PBP2) was also evaluated by nuclear magnetic resonance (NMR) spectroscopy and scanning electron microscopy (SEM). Transcriptomic sequencing and bioinformatic analyses were performed to elucidate global gene expression changes of <em>A. baumannii</em> BAA-1605 following exposure to meropenem and/or EGCG.</div></div><div><h3>Results</h3><div>Green tea aqueous extract synergized with meropenem against <em>A. baumannii</em> BAA-1605 by lowering the minimum inhibitory concentration (MIC) by 4-fold (128 to 32 μg/mL). EGCG was determined as the primary synergistic component in the extract (FICI = 0.375). The mechanism study showed that EGCG does not influence β-lactamase production or meropenem-mediated PBP2 inhibition but contributes to overcoming resistance by inhibiting efflux pump activity. Furthermore, transcriptomic analysis revealed that EGCG may suppress the AdeABC efflux system and enhance meropenem influx via a YiaD-dependent porin pathway.</div></div><div><h3>Conclusion</h3><div>This study presents a dual role of efflux inhibition and influx facilitation by EGCG in restoring meropenem efficacy, supporting its potential as a promising adjuvant to restore β-lactam potency against <em>A. baumannii</em> or other Gram-negative bacteria harbouring homologs of AdeABC and YiaD proteins.</div></div>","PeriodicalId":20212,"journal":{"name":"Phytomedicine","volume":"152 ","pages":"Article 157871"},"PeriodicalIF":8.3,"publicationDate":"2026-01-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146080634","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2026-01-23DOI: 10.1016/j.phymed.2026.157876
Fang Zhang , Haonan Xu , Yan Zuo , Ke Che , Yu Cui , Zhenhua Niu , Weiliang Cao , Tingting Sun , Yan Che , Hao Yu , Hao Chen
Background
Oligopeptides derived from dietary sources are regarded as ideal functional ingredients for nutritional interventions in diabetes due to their favorable bioavailability, target specificity, and safety profiles. Gynura divaricata (GD), a medicinal food plant, has shown hypoglycemic properties; however, the potential of GD-derived oligopeptides in glycemic control and their mechanistic underpinnings remain largely unexplored.
Purpose
This study aimed to investigate the anti-diabetic efficacy of GD oligopeptides and elucidate their mechanisms of action, particularly via the gut-microbiota-brain axis, in a streptozotocin (STZ)/high-fat diet-induced diabetic mouse model.
Methods
The therapeutic effects of GD oligopeptides were assessed through longitudinal blood glucose monitoring and systemic biochemical profiling. Organ-specific protection was evaluated via histopathological examination of the liver, pancreas, intestine, and brain. The influence on gluconeogenesis was analyzed by quantifying key glycogen metabolic proteins. Gut microbiota composition was assessed by sequencing, short-chain fatty acids (SCFAs) were measured, and appetite/energy metabolism regulators in the brain were detected. And a subset of diabetic mice was subjected to broad-spectrum antibiotic treatment to validate the effcst of gut microbes. Bioactive peptides were identified using LC-ESI-MS/MS, and molecular docking was performed to evaluate binding affinity to AKT.
Results
GD oligopeptides significantly ameliorated hyperglycemia, dyslipidemia, and insulin resistance in diabetic mice. They enhanced hepatic glycogen synthesis and suppressed gluconeogenesis through activation of the AKT/FoxO1 pathway, and reduced pancreatic apoptosis via modulation of Bcl-2/Bax expression. A total of 37 bioactive peptides were identified, with molecular docking confirming strong binding between GD oligopeptides and AKT. Importantly, GD oligopeptides restored intestinal barrier integrity, enriched SCFA-producing Lachnospiraceae, and promoted GPR43-dependent GLP-1 secretion, leading to hypothalamic GLP-1R activation, subsequent POMC upregulation, and NPY/AgRP suppression, collectively normalizing energy homeostasis. Crucially, all these therapeutic benefits were substantially reduced following antibiotic-induced microbiota depletion.
Conclusion
These findings underscore the potential of GD oligopeptides as a novel functional food ingredient for diabetes management by simultaneously targeting gluconeogenesis, gut microbiota, and central energy regulation, providing a mechanistic foundation for clinical translation.
{"title":"Oligopeptides from Gynura divaricata improve glycemic control via inhibition of gluconeogenesis and gut-brain axis regulation","authors":"Fang Zhang , Haonan Xu , Yan Zuo , Ke Che , Yu Cui , Zhenhua Niu , Weiliang Cao , Tingting Sun , Yan Che , Hao Yu , Hao Chen","doi":"10.1016/j.phymed.2026.157876","DOIUrl":"10.1016/j.phymed.2026.157876","url":null,"abstract":"<div><h3>Background</h3><div>Oligopeptides derived from dietary sources are regarded as ideal functional ingredients for nutritional interventions in diabetes due to their favorable bioavailability, target specificity, and safety profiles. Gynura divaricata (GD), a medicinal food plant, has shown hypoglycemic properties; however, the potential of GD-derived oligopeptides in glycemic control and their mechanistic underpinnings remain largely unexplored.</div></div><div><h3>Purpose</h3><div>This study aimed to investigate the anti-diabetic efficacy of GD oligopeptides and elucidate their mechanisms of action, particularly via the gut-microbiota-brain axis, in a streptozotocin (STZ)/high-fat diet-induced diabetic mouse model.</div></div><div><h3>Methods</h3><div>The therapeutic effects of GD oligopeptides were assessed through longitudinal blood glucose monitoring and systemic biochemical profiling. Organ-specific protection was evaluated via histopathological examination of the liver, pancreas, intestine, and brain. The influence on gluconeogenesis was analyzed by quantifying key glycogen metabolic proteins. Gut microbiota composition was assessed by sequencing, short-chain fatty acids (SCFAs) were measured, and appetite/energy metabolism regulators in the brain were detected. And a subset of diabetic mice was subjected to broad-spectrum antibiotic treatment to validate the effcst of gut microbes. Bioactive peptides were identified using LC-ESI-MS/MS, and molecular docking was performed to evaluate binding affinity to AKT.</div></div><div><h3>Results</h3><div>GD oligopeptides significantly ameliorated hyperglycemia, dyslipidemia, and insulin resistance in diabetic mice. They enhanced hepatic glycogen synthesis and suppressed gluconeogenesis through activation of the AKT/FoxO1 pathway, and reduced pancreatic apoptosis via modulation of Bcl-2/Bax expression. A total of 37 bioactive peptides were identified, with molecular docking confirming strong binding between GD oligopeptides and AKT. Importantly, GD oligopeptides restored intestinal barrier integrity, enriched SCFA-producing Lachnospiraceae, and promoted GPR43-dependent GLP-1 secretion, leading to hypothalamic GLP-1R activation, subsequent POMC upregulation, and NPY/AgRP suppression, collectively normalizing energy homeostasis. Crucially, all these therapeutic benefits were substantially reduced following antibiotic-induced microbiota depletion.</div></div><div><h3>Conclusion</h3><div>These findings underscore the potential of GD oligopeptides as a novel functional food ingredient for diabetes management by simultaneously targeting gluconeogenesis, gut microbiota, and central energy regulation, providing a mechanistic foundation for clinical translation.</div></div>","PeriodicalId":20212,"journal":{"name":"Phytomedicine","volume":"153 ","pages":"Article 157876"},"PeriodicalIF":8.3,"publicationDate":"2026-01-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146192188","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2026-01-23DOI: 10.1016/j.phymed.2026.157873
Xiaoyao Ma , Yang Zhang , Lishan Sun , Hongwei Zhao , Zihan Wang , Cui Hao , Wei Wang
Background
Influenza A virus (IAV) poses a serious threat to human health, and the increasing problem of drug resistance, along with the emergence of highly pathogenic strains, makes the development of new antiviral drugs urgent. Neferine, a dibenzylisoquinoline alkaloid derived from Nelumbo nucifera Gaertn plant, possesses multiple pharmacological activities, including anti-tumor, cardiovascular protection, neuroprotection, and anti-inflammatory effects.
Purpose
In this study, the anti-influenza A virus (IAV) activities and mechanisms of Neferine in vitro and in vivo were investigated in order to provide reference for the development of novel plant-derived anti-IAV drugs.
Methods
The antiviral activity of Neferine against IAV in vitro was evaluated using plaque reduction assay, RT-PCR, and western blot assay. The anti-IAV mechanisms of Neferine's were determined through mini-genome assay, DARTS assay, and SPR analysis. The in vivo anti-IAV effects of Neferine were investigated using a mouse pneumonia model combined with HE staining.
Results
Neferine exhibits broad-spectrum and highly effective antiviral activity against IAV at the cellular level. Neferine can "trap" the influenza virus in early endosomes, preventing its transport from early to late endosomes. Neferine primarily targets the NP protein of IAV to block its nuclear import and oligomerization. Neferine may bind to the Glu339 site of NP, which is a highly conserved. Additionally, it demonstrates significant protective effects against IAV infection in mice, notably improving the survival rates and accelerating weight recovery.
Conclusion
Neferine exhibits significant anti-IAV activity both in vitro and in vivo. It inhibits IAV infection by blocking the transport of IAV from early to late endosomes and inhibiting the functions of nucleoprotein (NP). These findings provide a substantial theoretical and experimental foundation for the development of Neferine as a novel anti-IAV agent.
{"title":"Inhibition of influenza a virus infection by natural isoquinoline alkaloid neferine targeting virus nucleoprotein","authors":"Xiaoyao Ma , Yang Zhang , Lishan Sun , Hongwei Zhao , Zihan Wang , Cui Hao , Wei Wang","doi":"10.1016/j.phymed.2026.157873","DOIUrl":"10.1016/j.phymed.2026.157873","url":null,"abstract":"<div><h3>Background</h3><div>Influenza A virus (IAV) poses a serious threat to human health, and the increasing problem of drug resistance, along with the emergence of highly pathogenic strains, makes the development of new antiviral drugs urgent. Neferine, a dibenzylisoquinoline alkaloid derived from <em>Nelumbo nucifera Gaertn</em> plant, possesses multiple pharmacological activities, including anti-tumor, cardiovascular protection, neuroprotection, and anti-inflammatory effects.</div></div><div><h3>Purpose</h3><div>In this study, the anti-influenza A virus (IAV) activities and mechanisms of Neferine <em>in vitro</em> and <em>in vivo</em> were investigated in order to provide reference for the development of novel plant-derived anti-IAV drugs.</div></div><div><h3>Methods</h3><div>The antiviral activity of Neferine against IAV <em>in vitro</em> was evaluated using plaque reduction assay, RT-PCR, and western blot assay. The anti-IAV mechanisms of Neferine's were determined through mini-genome assay, DARTS assay, and SPR analysis. The <em>in vivo</em> anti-IAV effects of Neferine were investigated using a mouse pneumonia model combined with HE staining.</div></div><div><h3>Results</h3><div>Neferine exhibits broad-spectrum and highly effective antiviral activity against IAV at the cellular level. Neferine can \"trap\" the influenza virus in early endosomes, preventing its transport from early to late endosomes. Neferine primarily targets the NP protein of IAV to block its nuclear import and oligomerization. Neferine may bind to the Glu339 site of NP, which is a highly conserved. Additionally, it demonstrates significant protective effects against IAV infection in mice, notably improving the survival rates and accelerating weight recovery.</div></div><div><h3>Conclusion</h3><div>Neferine exhibits significant anti-IAV activity both <em>in vitro</em> and <em>in vivo</em>. It inhibits IAV infection by blocking the transport of IAV from early to late endosomes and inhibiting the functions of nucleoprotein (NP). These findings provide a substantial theoretical and experimental foundation for the development of Neferine as a novel anti-IAV agent.</div></div>","PeriodicalId":20212,"journal":{"name":"Phytomedicine","volume":"152 ","pages":"Article 157873"},"PeriodicalIF":8.3,"publicationDate":"2026-01-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146080630","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
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