The FASEB Journal最新文献

Computer-aided drug discovery (CADD) has revolutionized the way we screen huge libraries of synthetic and natural compounds, offering a faster and more affordable alternative to traditional lab-based assays. Yet, questions remain about the reliability of these computational predictions, especially given the risk of false positives that cast uncertainty over the entire screening process. To explore this, we tested a library of 354 flavonoids against caspase-3; a key enzyme involved in apoptosis, and compared the results from different docking tools with in vitro enzymatic inhibition assays. Our objectives were threefold: to identify a highly reliable docking tool for screening flavonoids, to identify potent caspase-3 inhibitors, and to determine whether these compounds could protect liver cells in a fatty liver disease model. Although AutoDock4 performed better than the other two tools in our study, the predictive accuracy of all three docking platforms did not correlate well with the biochemical screen. While consensus strategies (RbR and RbE) improved ranking accuracy, they showed limited reliability as well. We identified six lead compounds that inhibited the caspase-3 activity. Out of which, in both cell lines (HepG2 & Huh-7), Okanin showed significant protection against amiodarone, while Okanin, Irigenin, and Isoliquiritigenin provided substantial protection against oleic acid-induced cell death. The findings highlight the limitations of individual docking programs and emphasize the utility of an integrated approach to enhance the effectiveness of predictions for virtual screening. Furthermore, this study provides a robust framework for the rational design of flavonoid-based caspase inhibitors with optimized efficacy and pharmacological profiles.

Moxifloxacin, a fluoroquinolone antibiotic with immunomodulatory activity, has not been evaluated for effects on autoantibody-driven inflammation. Pemphigoid diseases are a group of autoimmune blistering skin diseases driven by pathogenic autoantibodies and granulocytes. With current treatments for pemphigoid diseases associated with substantial adverse effects, there is a high medical need for new treatment strategies. We investigated the effect of moxifloxacin on skin inflammation in the antibody transfer mouse model of bullous pemphigoid-like epidermolysis bullosa acquisita. Moxifloxacin attenuated skin inflammation in this model markedly. In vitro, moxifloxacin inhibited responses of neutrophils to fixed IgG immune complexes, including the release of leukotriene B₄ (LTB₄), a lipid mediator essential for disease propagation in this model. Moxifloxacin also reduced the maximal respiration rate of mitochondria and inhibited mitochondrial complex I. Consistent with a hypothetical direct link between mitochondrial complex I actions and the release of LTB₄, the mitochondria-targeted antioxidant 10-(6′-plastoquinonyl)-decyltriphenylphosphonium (SkQ1; visomitin) reduced the immune complex-induced release of LTB₄ from neutrophils dose-dependently. Collectively, our results demonstrate that moxifloxacin can ameliorate autoantibody-induced granulocytic skin inflammation. The disruption of select mitochondrial functions and of the immune complex-induced release of LTB₄ from neutrophils might contribute to these therapeutic effects. Hence, moxifloxacin should be assessed as a drug for the treatment of patients with pemphigoid diseases.

BackgroundInflammatory bowel disease (IBD) pathogenesis involves immune dysfunction and gut microbiota dysbiosis. Aucubin (AU), a naturally occurring iridoid glycoside known for its ability to alleviate inflammation and modulate intestinal flora, has not yet been investigated in the context of colitis MethodsThe effects of AU on DSS-induced colitis in mice were evaluated, including disease severity, the balance of regulatory T cells (Tregs) and Th17 cells, intestinal barrier, inflammatory markers, fecal short-chain fatty acids (SCFAs), and safety. The gut microbiota was assessed using 16S rRNA sequencing. GPR41/43 involvement was tested using receptor antagonists. The antibiotic-treated mice and fecal transplantation were used to validate the microbiota-dependent effects. ResultsAU treatment broadly ameliorated the DSS-induced colitis and restored immune homeostasis by rebalancing Treg/Th17 cell populations. Meanwhile, intestinal barrier function was reinforced through upregulation of MUC2 and enhanced tight junction protein levels. Importantly, AU modulated gut microbiota composition, particularly enriching taxa associated with SCFA production, which indeed led to elevated fecal SCFA levels. Interestingly, the therapeutic effects of AU were absent in microbiota-depleted mice but could be conferred to DSS-induced recipients via fecal microbiota transplantation from AU-treated donors. Furthermore, the protective benefits of AU were partially attenuated upon pharmacological inhibition of the SCFA receptors GPR41/GPR43. No treatment-related toxicity was observed. ConclusionsAU safely alleviates colitis in mice by rebalancing the gut microbiota, activating SCFAs-GPR41/GPR43 axis to support mucosal immune regulation and barrier repair.

Acute kidney injury (AKI) is a serious complication of cisplatin chemotherapy, requiring new preventive therapies. Germacrone, a sesquiterpenoid derived from Rhizoma Curcuma, has multiple pharmacological effects, including anti-inflammatory, antiapoptotic, and antioxidant effects. The current study aimed to investigate the effect and mechanism of germacrone on AKI. Single high-dose and multiple low-dose cisplatin-induced AKI and AKI-chronic kidney disease (CKD) transition mouse models were used to determine the preventive effect of germacrone, with the mechanism explored in HK-2 and Raw264.7 cells. A breast cancer tumor-bearing model was developed to determine whether the nephron-protective effects of germacrone affect the anticancer properties of cisplatin. Here, we found that germacrone significantly alleviated AKI and fibrosis in cisplatin-treated mice, as demonstrated by decreased serum creatinine and blood urea nitrogen (BUN) levels and improved renal pathology. Germacrone attenuated cisplatin-induced apoptosis and inflammation in vivo and in vitro. RNA sequencing revealed that germacrone restored cisplatin-induced gene dysregulation and regulated inflammation and apoptosis through the PI3K/AKT and Ras/MAPK pathways. Inhibition of these pathways abrogated the renoprotective effect of germacrone in vitro. Moreover, germacrone reduced macrophage activation and induced an M2-dominant shift in macrophage polarization in the kidneys and Raw264.7 cells via similar pathways. Furthermore, germacrone protected the kidneys without affecting the chemotherapeutic effects of cisplatin on a mouse model of breast cancer. Therefore, germacrone is expected to have universal renoprotective effects across different AKI models and a tumor-bearing model through similar signaling pathways, suggesting its potential as a clinical adjuvant therapy for reducing nephrotoxicity in patients receiving cisplatin chemotherapy.

Sepsis-associated acute kidney injury (SA-AKI) is a significant form of organ dysfunction that affects a substantial proportion of patients with sepsis, contributing to increased mortality. However, the potential mechanisms underlying SA-AKI remain poorly understood. In this study, we identified Rab27a as a critical factor in regulating SA-AKI, with its upregulated expression both in vitro and in vivo. The silence of Rab27a mitigated LPS-induced injury and mitochondrial dysfunction in HK-2 cells, leading to enhanced cell migration and functional improvement. Notably, the protective effects of Rab27a knockdown were reversed upon silencing PPAR-γ, highlighting a potential correlation between these proteins. Furthermore, a significant interaction between PPAR-γ and AMPK-α1 was observed following Rab27a knockdown in RTECs during SA-AKI. In conclusion, we demonstrated that Rab27a served as a key mediator in the pathogenesis of SA-AKI, influencing RTEC function and mitochondrial integrity.

Thoracic aortic dissection (TAD) is a life-threatening vascular pathology with limited therapeutic options. The role of the small molecule STF083010, a novel IRE1α RNase-specific inhibitor, in TAD has not yet been defined. We aimed to target the IRE1α-XBP1s axis pharmacologically using STF083010, and evaluate its therapeutic potential in the occurrence and progression of TAD. Using a β-aminopropionitrile monofumarate (BAPN)-induced murine TAD model combined with interleukin 1β (IL1β)-stimulated vascular smooth muscle cells (VSMCs), we systematically evaluated the effects of STF083010 on the development of TAD as well as VSMCs' phenotypic switching and vascular inflammation. Our study reveals that STF083010 administration significantly attenuates TAD formation in vivo. Furthermore, we identified STF083010 confers VSMCs with resistance to IL1β-induced VSMCs inflammation and phenotypic switching. Mechanistically, STF083010 restrains IRE1α-XBP1s axis and suppresses NLRP3 inflammasome activation-dependent pyroptosis, further inhibiting VSMCs phenotypic switching and inflammatory activation, and eventually alleviates the progress of TAD. Our findings establish STF083010 as a potential molecule drug for the prevention of TAD and provide mechanistic insights for developing innovative therapies in aortic pathologies.