Background Atherothrombosis is a chronic, multifactorial, and progressive disease responsible for a significant number of deaths globally. It involves interactions between endothelial dysfunction, coagulation activation, platelet activity, and fibrin turnover. Combined oral contraceptives (COCs) may influence these pathways, potentially altering thrombotic risk. The aim was to assess the impact of COC administration on metabolic status and the risk of atherothrombotic disorders in young female rats, beyond evaluating the therapeutic effects of short-term low-dose aspirin (LDA) treatment. Methods Thirty (n = 30) five-week-old female Sprague Dawley rats were given low-dose COC (LCOC) or a high-dose COC (HCOC) for six weeks before treatment with LDA for another four weeks. These rats were compared with those given only LCOC or HCOC without LDA treatment. Whereas rats that received distilled water (as a vehicle) and LDA only served as controls. Body weight and metabolic status were measured weekly. Parameters related to glucose regulation, lipid profiles, inflammatory cytokines, haematological indices, coagulation, and endothelial dysfunction were evaluated at the end of the experiment. Results Rats exposed to HCOC exhibited abnormal metabolic and lipid profiles, including impaired glucose tolerance, accompanied by significantly higher insulin (p < 0.001), triglycerides and very low-density lipoprotein levels (p = 0.004) when compared with control groups. The HCOC treatment was also associated with elevated platelet count (p = 0.02) and markers of inflammation and endothelial dysfunction, including IL-6 (p < 0.05), TNF-alpha (p < 0.05), MCP-1 (p < 0.001), TF (p < 0.001), D-dimer (p < 0.001), von Willebrand factor (p < 0.05), and low nitric oxide (p < 0.001) when compared with control groups. Notably, SBP and MAP (p = 0.02) were also significantly higher in these animals, suggesting an increased atherothrombotic risk in response to HCOC when compared to control groups. Interestingly, short-term LDA attenuate atherothrombotic risk by improving metabolic status, alleviating inflammation, and lowering the markers of coagulation and endothelial dysfunction in rats exposed to HCOC. Conclusion Short-term LDA reduces the risk of atherothrombotic disorders by attenuating biomarker level associated with metabolic dysregulation, endothelial dysfunction, proinflammation, and hypercoagulation in female rats exposed to high-dose COC. However, additional experimental studies are required to explore the mechanistic interaction between LDA and COC at the cellular level.
The working heart–brainstem preparation (WHBP) is an arterially perfused in situ rodent model that requires an osmotically or oncotically active perfusate additive to limit edema and maintain stable central respiratory motor output. Ficoll-70 at 1.25% (w/v) is widely used for this purpose, but at this concentration it generates only modest colloid osmotic pressure (∼3–4 mmHg) and presents practical limitations, including batch variability, endotoxin risk, and high cost. We asked whether mannitol at 0.85% (w/v), a small-molecule crystalloid osmolyte and hydroxyl-radical scavenger with an established clinical safety profile, can substitute for Ficoll-70 while preserving WHBP viability.Adult mouse WHBPs (n = 7 Ficoll; n = 9 mannitol) were perfused for 1 h with Krebs-based perfusate containing either 1.25% Ficoll-70 or 0.85% mannitol. Preparations were assessed for phrenic and vagus nerve activity, perfusion pressure, whole-preparation weight gain, brain tissue weight, and single-neuron recording capability. Phrenic discharge frequency increased over time in both groups (Ficoll: 0.15 → 0.49 Hz; mannitol: 0.24 → 0.45 Hz), and amplitude of the integrated phrenic burst did not differ significantly between additives. Perfusion pressure rose modestly over 1 h (Ficoll: 67.7 → 76.7 mmHg; mannitol: 67.2 → 74.1 mmHg), with no significant between-group difference. Whole-preparation weight increased in both conditions, indicating tissue water uptake during perfusion. Brainstem–cerebellum wet weight after 1 h was similar with both perfusates (0.140 ± 0.023 g, Ficoll; 0.124 ± 0.020 g, mannitol; p = 0.18). Juxtacellular recordings from preBötzinger complex neurons confirmed that single-unit activity can be obtained reliably in mannitol-perfused preparations for more than 2 h. Moreover, post-inspiratory activity of the vagus nerve is preserved in mannitol-perfused preparations.Together, these data indicate that 0.85% mannitol maintains WHBP cardiorespiratory function and tissue water balance to a degree comparable with 1.25% Ficoll-70, while providing a lower-cost, endotoxin-free alternative. The use of mannitol as a crystalloid perfusate additive may therefore facilitate wider and more reproducible application of the WHBP and related arterially perfused in situ preparations in integrative physiology and pharmacology.
Diabetic kidney disease (DKD) is the leading cause of end-stage renal disease (ESRD), but early diagnosis remains challenging. Current reliance on indicators such as urinary albumin and glomerular filtration rate is limited by insufficient sensitivity and susceptibility to interference. The pathogenesis of DKD is complex, involving multiple intersecting pathways and epigenetic modifications in the progression of the disease. Omics biomarkers offer new directions for early risk assessment, while machine learning can integrate multi-omics data to build efficient diagnostic models. This article reviews recent advances in biomarker research, discusses strategies for model construction, and provides theoretical references for early and precise diagnosis and clinical translation of DKD.
