The FASEB Journal最新文献

Dysbiosis of the gut microbiota in diabetes is accompanied by reduced levels of short-chain fatty acids (SCFAs), including butyrate, a four-carbon SCFA with immunomodulatory activity. Inflammatory stimulation drives macrophage polarization toward an M1 phenotype and can induce concurrent activation of markers associated with pyroptosis, apoptosis, and necroptosis, consistent with a PANoptosis-like cell death phenotype that may exacerbate periodontal destruction. Here, we investigated whether butyrate attenuates diabetic periodontitis by restraining macrophage M1 polarization and reducing PANoptosis-like cell death. In a mouse model of diabetic periodontitis, oral butyrate treatment alleviated alveolar bone loss and reduced M1 polarization and PANoptosis-like death in periodontal tissues. In THP-1–derived macrophages, we assessed inflammatory polarization and PANoptosis-like cell death under inflammatory stimulation with or without butyrate pretreatment. Butyrate suppressed M1-associated programs and reduced PANoptosis-like cell death, accompanied by inhibition of histone deacetylase 3 (HDAC3) and attenuation of signal transducer and activator of transcription 1 (STAT1) signaling. Moreover, butyrate mitigated inflammatory responses in periodontal ligament stem cells (PDLSCs) and promoted osteogenic differentiation. Collectively, these findings suggest that butyrate mitigates diabetic periodontitis progression by suppressing macrophage inflammatory programs while supporting PDLSC osteogenesis, highlighting its potential as an adjunctive immunomodulatory approach.

Inflammation and oxidative stress play crucial roles in the pathogenesis of diabetic kidney disease (DKD). Hirudin, a small molecular polypeptide derived from the salivary glands of leeches, is widely utilized in anti-coagulation and antithrombotic therapies. However, the effects and underlying molecular mechanisms of hirudin on DKD remain unclear. Db/db mice were employed to evaluate the effects of hirudin on DKD. Key parameters assessed included urinary albumin, oral glucose tolerance, glomerular diameter, and the expression levels of NLRP3, IL-1β, IL-18, caspase-1, and reactive oxygen species (ROS). Additionally, proteomic analysis was performed to measure the β-hydroxybutyrylation level of SOD₂, and the effects of changes in SOD₂ β-hydroxybutyrylation were evaluated by immunoprecipitation. In vivo experiments demonstrated that hirudin significantly improved urinary albumin levels, oral glucose tolerance, and glomerular diameter in diabetic mice. Furthermore, the β-hydroxybutyrylation level of SOD₂ was reduced, leading to decreased production of ROS and suppression of NLRP3 inflammasome activation. In vitro experiments indicated that hirudin reduced the polarization of RAW264.7 cells, lowered their ROS levels, diminished NLRP3 inflammasome activation, and reduced the β-hydroxybutyrylation modification level of SOD₂. Hirudin can alleviate the progression of DKD by reducing the β-hydroxybutyrylation level of SOD₂, which in turn reduces ROS production and NLRP3 inflammasome activation, thereby suppressing inflammation. These findings provide new insights into the potential application of hirudin in the context of DKD.

Progressive peritoneal fibrosis is a severe complication of peritoneal dialysis (PD) with a complex pathogenesis. Our previous studies demonstrated that parthenolide (PTL) alleviates peritoneal fibrosis by suppressing the transforming growth factor (TGF)-β/Smad pathway. Smad anchor for receptor activation (SARA) acts as an adaptor protein for Smad2 and Smad3; however, its role in PD-associated peritoneal fibrosis and the relationship between PTL and SARA remain to be elucidated. In this study, single-cell sequencing (scRNA-seq) data and long-dwell PD fluid samples were collected. PD mice, a TGF-β1-induced mesothelial–mesenchymal transition (MMT) model, and CRISPR/Cas9-engineered SARA gene (ZFYVE9) knockout MeT-5A cells were established. The results revealed that SARA activates Smad2/3, whereas Smad3 promotes SARA degradation, leading to the downregulation of SARA expression during fibrosis progression in long-term-retained PD fluid samples, PD mice, TGF-β1-treated MeT-5A cells, and HMrSV5 cells. PTL does not regulate SARA protein stability, but luciferin reporter gene experiments revealed that the transcriptional recovery of the effects of PTL on SARA is consistent with its known antifibrotic effects. PTL was further confirmed to inhibit the SARA-Smad3 interaction through both in vitro and in vivo coimmunoprecipitation experiments. Molecular docking and PTL-biotin pulldown assays confirmed that PTL directly binds to the Smad3-binding interface of SARA at the Pro788 and Ser795 residues. In summary, SARA promotes PD-related peritoneal fibrosis through the phosphorylation of Smad2/3. PTL specifically binds to ZFYVE9 (P788/S795), disrupting the interaction between SARA and Smad3, thereby improving MMT and alleviating peritoneal fibrosis, which in turn restores SARA expression. This study provides a theoretical foundation for the clinical diagnosis and treatment of peritoneal fibrosis.

Cardiovascular interventional therapy continues to face a major challenge in clinical practice due to the presence of restenosis after PCI. Continuous exploration uncovers novel signaling molecules implicated in this pathophysiological process. However, the precise molecular mechanism remains elusive. The WW domain-binding protein 2 (WBP2) has emerged as a notable oncoprotein, serving as a central hub that links multiple signaling pathways in cancer, including EGFR, PI3K, Hippo, and Wnt. Nevertheless, its role in vascular biology remains ambiguous. This study aims to elucidate the role of WBP2 in neointimal hyperplasia (NIH) as well as vascular smooth muscle cell (VSMC) proliferation after vascular injury. The mice carotid artery ligation (CAL) model revealed increased WBP2 expression after vascular injury, which was further confirmed by PDGF-BB stimulation in VSMCs. Histopathological analysis was performed to assess the extent of NIH in the CAL mouse model. Additionally, FUCCI and Transwell assays were used to evaluate VSMC proliferation and migration, respectively. WBP2 knockdown alleviated neointimal thickening and VSMC proliferation following vascular injury, in stark contrast to the significant increase observed with WBP2 overexpression. Mechanistically, we demonstrated that WBP2 interacts with Y-box binding protein 1 (YBX1) and enhances the binding of RSK to YBX1, promoting YBX1 S102 phosphorylation, which facilitates its nuclear translocation. This subsequently activates proliferative genes and represses contractile genes, thereby contributing to the development of NIH. Our findings suggest that WBP2 may promote NIH and VSMC proliferation by facilitating the nuclear translocation of YBX1. Therefore, WBP2 could serve as a promising target for the management of restenosis.

Colorectal cancer (CRC) remains a major cause of cancer-related morbidity and mortality worldwide, underscoring the need to identify new drivers of tumor progression. HAUS augmin-like complex subunit 1 (HAUS1), a component of the microtubule–augmin complex, has been implicated in mitotic spindle assembly; however, its clinical significance and mechanistic contribution to CRC are largely unknown. Here, we reported that HAUS1 was markedly upregulated in CRC tissues and cell lines, and its high expression correlated with lymphatic metastasis and poor patient prognosis. Functional experiments demonstrated that HAUS1 depletion significantly impaired CRC cell proliferation, migration, and tumor growth in vitro and in vivo. Mechanistically, transcriptomic profiling and integrative bioinformatic analyses identified CDCA8 as a key downstream effector of HAUS1. HAUS1 physically interacted with EZH2 and facilitated the recruitment of E2F1 to the CDCA8 promoter, thereby enhancing CDCA8 transcription through a methylation-independent HAUS1–EZH2–E2F1 axis. Disruption of CDCA8 abrogated the oncogenic effects induced by HAUS1 overexpression, confirming its essential role in HAUS1-driven tumor progression. Furthermore, HAUS1 was required for maintaining cancer stem-like properties, as HAUS1 silencing reduced stemness marker expression, impaired sphere formation, and decreased tumor-initiating cell frequency in vivo—effects that were reversed by CDCA8 restoration. Collectively, our findings identified HAUS1 as a clinically relevant oncogenic driver that promoted CRC progression and cancer stemness by transcriptionally activating CDCA8. Targeting the HAUS1–EZH2–E2F1–CDCA8 signaling axis might represent a promising therapeutic strategy for CRC.

Oxidant injury to the respiratory epithelium is thought to underlie adverse effects of air pollution on respiratory health, with individual susceptibility likely to determine severity of adverse effects. Knowledge of individual susceptibility would facilitate selecting appropriate participants into randomized clinical trials (RCTs) of antioxidant strategies. Growing primary nasal epithelial cells (NECs) into fully differentiated epithelium in air-liquid interface culture, we separated subjects into sensitive or resistant to oxidant injury by the concentration of H₂O₂ required to induce epithelial leak (FITC-dextran leak > 10 μg/mL). Subjects were classified into sensitive (< 25 mM H₂O₂) or resistant (≥ 25 mM H₂O₂) to oxidative stress. However, 10–12 weeks required to obtain this result is too long for use in screening RCT participants. We previously demonstrated that sensitive participants also showed mitochondrial dysfunction at baseline and thus reasoned that measuring mitochondrial function in submerged monolayer cultures would allow more rapid subject classification. NECs from 38 healthy non-smoking adults had mitochondrial function measured using a Seahorse XF analyzer. K-means cluster analysis based on ATP production and basal respiration independently classified participants into sensitive and resistant groups. Full oxidant epithelial challenge confirmed with 100% classification agreement. Our new screening test provides results in 2–3 weeks and is more practical for screening potential participants in RCTs of antioxidant therapies.