Second-generation combined oral contraceptive pills (COCPs), particularly formulations containing ethinylestradiol and levonorgestrel, remain widely used because of their effectiveness and affordability in middle and low-income countries. However, concerns persist about their cardiovascular safety. This study investigated the dose-dependent effects of a second-generation COCP on systolic blood pressure (SBP), inflammatory biomarkers, oxidative stress, vascular mediators, and vascular histology in female Sprague–Dawley rats. Twenty-four female Sprague–Dawley rats were acclimatized and randomly assigned to control, low-dose (1.1 µg/kg/day), and high-dose COCP (11 µg/kg/day) groups. Treatments were administered orally for six weeks. SBP and body weight were monitored before and after treatment. Serum C-reactive protein (CRP), tumour necrosis factor-alpha (TNF-α), and lipid profile were measured. Renal vascular homogenates were analyzed for nuclear factor kappa B (NF-κB), endothelin-1 (ET-1), nitric oxide (NO), malondialdehyde (MDA), superoxide dismutase, catalase (CAT), and glutathione. Histological examination was performed on the renal and thoracic arteries. The SBP increased by 4.6
Musculoskeletal aging exhibits prominent tissue heterogeneity. Forkhead box O3 (FOXO3) acts as a master transcription factor orchestrating aging, metabolism and immune homeostasis. Existing mechanistic investigations are largely restricted to single tissues, and a head-to-head comparison of FOXO3-governed immunometabolic regulatory networks between knee osteoarthritis (KOA) synovium and aged skeletal muscle remains absent. A retrospective multi-cohort transcriptomic analysis was performed using datasets retrieved from the Gene Expression Omnibus (GEO) database. Two KOA synovial cohorts (GSE55235 and GSE55457, n = 40) and two aged skeletal muscle cohorts (GSE28422 and GSE25941, n = 146) were enrolled, covering a total of 186 tissue samples. We integrated differential expression analysis, random-effects meta-analysis, receiver operating characteristic (ROC) performance evaluation, single-sample gene set enrichment analysis (ssGSEA), and partial correlation analysis adjusted for disease confounding effects. A bootstrap-scoring system was further established to prioritize candidate therapeutic strategies. FOXO3 exhibited opposing expression patterns: it was downregulated in KOA synovium yet upregulated in aged skeletal muscle. The coupling profiles between FOXO3 and immune or metabolic pathways displayed striking tissue-specific divergence. In KOA synovium, FOXO3 expression was positively correlated with mitochondrial energy metabolism and macrophage infiltration, accompanied by selective inflammatory immune infiltration. By contrast, aged skeletal muscle resided in an immune-quiescent microenvironment, where elevated FOXO3 was strongly negatively coupled to oxidative metabolic pathways. Distinct intervention strategies were derived from these divergent network signatures: combined FOXO3 activation and metabolic modulation was recommended for KOA, whereas supplementation with nicotinamide adenine dinucleotide (NAD⁺) precursors constituted the primary intervention for aged skeletal muscle. Opposite FOXO3 expression levels correspond to fundamentally distinct immunometabolic network landscapes during musculoskeletal aging. The functional output of FOXO3 is dictated by the tissue microenvironment rather than its transcriptional abundance alone. This study supports stratified precision geriatric interventions via tissue-specific modulation of the FOXO3 pathway across divergent musculoskeletal aging contexts. Clinical trial number: Not applicable.
Rhodiola rosea L., a medicinal plant native to the high-altitude regions of the Indian Himalayas, contains diverse bioactive phytochemicals with broad pharmacological activities. This study investigated the anticancer activity and associated molecular responses of R. rosea root extracts in human breast cancer models. Commercial Rhodiola rosea root powder reconstituted in different solvents was evaluated for cytotoxicity against SK-BR-3, MDA-MB-231, and MCF-7 breast cancer cells, with HEK-293 cells serving as a non-malignant human reference model. The extracts exhibited concentration- and time-dependent cytotoxic effects, with differential sensitivity among breast cancer subtypes. MDA-MB-231 cells were subsequently selected for mechanistic investigations. Scratch wound-healing assays demonstrated reduced cell migration following extract treatment, accompanied by decreased expression of epithelial–mesenchymal transition (EMT)-associated proteins, including Snail, Slug, Vimentin, and N-Cadherin. Flow-cytometric analysis revealed extract-dependent redistribution of cells across the G0/G1, S, and G2/M phases, with dimethyl sulfoxide (DMSO)-solubilized R. rosea extract (RRD) and methanol-solubilized R. rosea extract (RRM) increasing the G0/G1 population and reducing the G2/M population, whereas ethanol-solubilized R. rosea extract (RRE) produced a distinct cell-cycle response. Gas chromatography–mass spectrometry (GC–MS) and liquid chromatography–mass spectrometry (LC–MS) profiling identified diverse phytochemical constituents in the extracts. Western blot analysis further demonstrated modulation of proteins involved in proliferation, cell-cycle regulation, apoptosis, tumor suppression, and EMT, including reduced hematopoietic PBX-interacting protein (HPIP)/protein kinase B (AKT) signaling, cellular myelocytomatosis oncogene (c-Myc), Cyclin A, Cyclin B, Fibrillarin (FBL), Aurora B, Snail, and Slug, together with increased phosphatase and tensin homolog (PTEN), phosphorylated retinoblastoma protein (pRb), AMP-activated protein kinase (AMPK), and cleaved Caspase-3. Collectively, these findings indicate that R. rosea extracts exert multifaceted anticancer effects through modulation of proliferative signaling, cell-cycle progression, apoptotic responses, and EMT-associated phenotypic changes.
Transcriptomic biological age clocks may track age-related transcriptional variation in skeletal muscle, but it remains unclear whether they detect sarcopenia-related molecular variation beyond chronological age. This multicohort study benchmarked directly implementable RNA-based age models in public human skeletal muscle RNA-seq datasets. GSE164471 was used as a healthy-aging reference, GSE111006/GSE111010 as a pooled discovery benchmark, GSE167186 as external validation, and GSE174106 as biological validation using appendicular lean mass trajectories. RNAAgeCalc skeletal muscle and across-tissue models were implemented for age estimation and sarcopenia discrimination. Gene coverage was used only as a preliminary technical-compatibility assessment, after which the published SkeletAge model was directly applied to GSE111006 and GSE111010 for independent chronological-age prediction and sarcopenia-discrimination analyses; GSE164471, which contributed to SkeletAge model development, was used only for an in-sample implementation check. Both RNAAgeCalc models showed high gene coverage. In GSE164471, the skeletal muscle model showed stronger chronological-age tracking than the across-tissue model (r = 0.604 vs. 0.266; MAE 15.2 vs. 16.7 years), but retained substantial absolute prediction error and positive mean bias (+ 10.4 years), indicating that stronger correlation did not imply adequate calibration. In the discovery benchmark, between-group differences in chronological age, transcriptomic age, and age-acceleration residuals were not statistically significant, with small-to-moderate standardized effect estimates and confidence intervals spanning zero. Logistic discrimination was influenced mainly by cohort origin, while transcriptomic aging measures provided limited independent discrimination. External validation showed broad age-group structure with substantial overlap between old healthy and sarcopenic participants, and no differences were detected across appendicular lean mass trajectories. Current RNAAgeCalc implementations therefore appear to track chronological-age-related transcriptomic structure more effectively than sarcopenia-related variation, although smaller disease-related effects cannot be excluded, and these models should not currently be considered standalone sarcopenia biomarkers.
Pathological cardiac hypertrophy induced by chronic β-adrenergic receptor (β-AR) stimulation contributes to adverse cardiac remodeling and the progression of heart failure, with NADPH oxidase (NOX2)-derived reactive oxygen species (ROS) acting as key mediators of redox-dependent hypertrophic signaling. Sustained ROS generation can promote oxidative activation of Ca²⁺/calmodulin-dependent protein kinase II (CaMKII) and downstream myocyte enhancer factor 2 C (MEF2C)-mediated transcriptional reprogramming. Polydatin, a natural glucoside of resveratrol, has been reported to exert antioxidant and cardioprotective effects. However, its influence on the NOX2–ROS–CaMKII–MEF2C axis under chronic β-AR stimulation remains unclear. In the present study, male Wistar rats were assigned to control, isoproterenol (5 mg/kg/day, subcutaneous, 4 weeks), or isoproterenol + polydatin (100 mg/kg/day, oral gavage) groups. Cardiac hypertrophy was evaluated by morphometric indices, ANP and BNP mRNA expression levels, and histological analyses. Oxidative stress was assessed through NADH oxidase activity, protein carbonyl content, and total antioxidant status. Protein expression of NOX2 subunits (gp91phox, p22phox, p47phox, p67phox, p40phox) and components of the CaMKII–MEF2C pathway were determined by Western blot and reverse transcription quantitative PCR (RT–qPCR). Isoproterenol administration induced marked cardiac hypertrophy, elevated oxidative stress markers, increased expression of NOX2 regulatory subunits, and activated the CaMKII–MEF2C pathway. Polydatin treatment significantly attenuated hypertrophic indices, reduced NADH oxidase activity and protein carbonyl levels, restored antioxidant capacity, and suppressed p47phox, p67phox, p40phox expression along with p-CaMKIIThr286/CaMKII, ox-CaMKII/CaMKII ratios and MEF2C expression. These findings suggest that the attenuation of isoproterenol induced cardiac hypertrophy by polydatin is associated with modulation of oxidative stress, NOX2 subunit expression, and CaMKII–MEF2C signaling, although a causal role of this pathway remains to be established.
