Against the background of active health management, the gut microbiota (GM) has been implicated as a potentially important factor associated with exercise-induced bodily responses. This study explored how complete exercise–environment protocols were associated with distinct physiological, microbial, metabolic, and immune responses under controlled thermal conditions, providing theoretical support for personalized exercise strategies and future microbiota-related research. Thirty-two healthy males (18–35 years) were assigned to four groups for a 10-day intervention: high-temperature/humidity equipment-assisted training (HE), normal-temperature/humidity equipment-assisted training (NE), the high-intensity interval training team at high temperature (HIIT), and a control group (Ctrl). Fecal and plasma samples were collected pre- and post-intervention, with GM, putatively annotated plasma metabolite features, and plasma cytokines detected by 16 S rRNA sequencing, untargeted metabolomics, and Luminex technology, followed by multi-omics correlation analysis. The HE, NE, and HIIT groups showed distinct GM compositional shifts: NE was associated with decreased Escherichia-Shigella and increased Blautia and Dorea relative abundance, HE was associated with decreased relative abundance of selected genera, and HIIT was associated with increases in selected genera. The three groups also presented characteristic metabolic and cytokine profiles, with exploratory within-group analyses reporting lower heat stress-induced CCL2 responses in HE/NE. Multi-omics analysis identified exploratory GM–plasma metabolite association patterns associated with exercise-related physiological responses. Under the specific combinations of session duration, intensity structure, and thermal exposure used here, the protocols showed descriptive physiological, microbial, metabolic, and cytokine response patterns. Operational HTT achievement was 0/8 in Ctrl, 7/8 in HE, 7/8 in NE, and 3/8 in HIIT. An exact four-group comparison indicated heterogeneity (p = 0.000163). The protocols were also associated with distinct exploratory gut microbiota, plasma metabolite-feature and cytokine patterns. These findings support protocol-specific physiological and multi-omics associations, but do not isolate the independent effect of exercise modality or establish causal microbiota–metabolite mechanisms.
Second generation antipsychotics (SGAs) are widely prescribed for the management of psychiatric disorders, yet their use is associated with obesity, insulin resistance, and impaired glucose homeostasis. Small extracellular vesicles (sEV) are mediators of inter-tissue communication in metabolic diseases. Herein, we investigated the impact of the SGAs aripiprazole (ARI) and olanzapine (OLA) on circulating sEV in healthy volunteers and patients with first-episode psychosis, as well as the effects of these sEV on hepatocyte insulin signalling and systemic inflammation. Twenty-four healthy volunteers received OLA or ARI for five days. Seven individuals with first-episode psychosis were treated with SGAs; three were followed longitudinally for 6 months. sEV were isolated from plasma or culture media of SGA-treated human macrophages (h-mac) from donors. Huh7 human hepatocytes were exposed to sEV and insulin signalling was assessed. Inflammatory responses were evaluated by determining IκBα or IL1B expression in h-mac. In healthy volunteers, ARI significantly increased circulating sEV levels, which positively correlated with LDH, GGT, and free T4 plasma levels. sEV from ARI-treated subjects impaired hepatocyte insulin signalling in most cases, whereas sEV from OLA-treated patients produced milder effects. sEV from SGA-treated h-mac induced hepatocyte insulin resistance, accompanied, particularly with ARI, by reduced IκBα and increased IL1B expression in h-mac. Patients showed elevated baseline circulating sEV and increased baseline IL1B expression in PBMCs compared to healthy volunteers, and SGAs further enhanced these alterations. Our results suggest that elevations in circulating sEV associated to disease and SGA treatment may contribute to insulin-desensitization and inflammation, two hallmarks of metabolic dysfunction. In healthy volunteers ARI induces a higher rise in circulating sEV than OLA Circulating sEV from ARI-treated volunteers attenuate hepatocyte insulin signalling sEV from ARI-treated h-mac increase IL1B expression and promote hepatocyte insulin resistance First-episode psychosis elevated plasma sEV and SGAs enhanced this effect
Lipotoxicity contributes to polyendocrine metabolic ovarian syndrome (PMOS) pathogenesis by inducing hyperandrogenism (HA) and insulin resistance (IR), while excess testosterone triggers endoplasmic reticulum (ER) stress and the unfolded protein response (UPR) in PMOS models. However, ER stress and UPR activation under HA-obese conditions remain unexplored in granulosa cells (GCs) and PMOS mouse liver and ovary. Neurokinin 3 receptor antagonists (NK3Ra) reverse PMOS metabolic and reproductive impairments; therefore we established an HA-obese model by exposing KGN cells to palmitic acid (PA) and dihydrotestosterone (DHT), and an HFD-letrozole PMOS mouse model, both treated with NK3Ra. In KGN cells, ER and mitochondrial ultrastructure disruption was observed by TEM, UPR sensor protein and gene expression/phosphorylation was analyzed by qPCR and western blotting, and NK3Ra rescue of cell viability was evaluated, alongside peripheral and tissue biochemical/hormonal assays, immunohistochemistry for AMH, UPR sensor and cascade proteins, and redox metabolism assays. NK3Ra alleviated ER stress by modulating the PERK–ATF4–CHOP axis, improving steroidogenesis, redox homeostasis, and viability in HA-obese models. In PMOS mice, hepatic steatosis was reversed with associated downregulation of CHOP, decreased de novo lipogenic gene expression, and increased superoxide dismutase activity, while estrous cyclicity and steroid hormone metabolism were restored, accompanied by reduced ovarian ATF4, CHOP, and GRP78 expression by NK3Ra activity. Collectively, our findings suggest that NK3Ra alleviates lipotoxic-HA-induced ER stress via the modulation of the PERK–ATF4–CHOP pathway, thereby ameliorating metabolic and endocrine aberrations in PMOS.
Arterial hypertension and diabetes mellitus frequently coexist and synergistically accelerate adverse left ventricular remodeling through mitochondrial dysfunction, sterile inflammation, and extracellular matrix disorganization. Heat shock protein 60 (HSP60) is a mitochondrial chaperone involved in proteostasis and stress adaptation, but it remains unclear whether cardiomyocyte-restricted augmentation of HSP60 can modulate myocardial remodeling and influence systemic immune-cell bioenergetics. Male spontaneously hypertensive rats (SHR) and normotensive Wistar–Kyoto (WKY) rats were studied with or without streptozotocin-induced diabetes. Animals were assigned to WKY, SHR, WKY+HSP60, SHR+HSP60, WKY + DM+HSP60, and SHR + DM+HSP60 groups. HSP60 was delivered using an adeno-associated virus serotype 9 (AAV9) vector under the cardiac troponin T promoter. Myocardial remodeling was assessed by histomorphometry and Masson’s trichrome staining, myocardial signaling by immunohistochemistry and RT-qPCR, and peripheral blood mononuclear cell (PBMC) mitochondrial membrane potential by JC-1 flow cytometry. Cardiac HSP60 delivery showed a context-dependent pattern. In hypertensive diabetic rats, this pattern included altered NLRP3 subcellular localization, modulation of mitochondrial regulatory and inflammatory-fibrotic transcripts, and increased PBMC mitochondrial membrane potential. PBMC polarization inversely correlated with myocardial fibrosis. Normotensive and single-pathology groups did not show a comparable integrated response profile. These findings indicate that cardiomyocyte-targeted HSP60 overexpression is associated with remodeling-related changes in myocardium and peripheral immune-cell bioenergetics under combined hypertensive and diabetic stress. PBMC mitochondrial membrane potential may represent a candidate peripheral indicator of myocardial state in this preclinical setting.
Pancreatic acinar cell pyroptosis played a key role in acute pancreatitis (AP), while the underlying molecular mechanisms were unclear. In this study, hyperlipidemic acute pancreatitis (HLAP) model mice and primary acinar cells were employed for the in vivo and in vitro experiments, respectively. Western blot was employed to assess Gasdermin D-N-terminal pore-forming domain (GSDMD-N) oligomerization; Flow cytometry was employed to assess cell pyroptosis; Immunofluorescence was performed to assess GSDMD-N location in the cytoplasm; HE and TUNEL staining were employed to assess the pathological changes and cell apoptosis of pancreatic tissues. Palmitic acid levels were elevated in serum and pancreatic tissues of HLAP mice, palmitoyltransferase Zinc Finger DHHC Domain-Containing Protein 13 (ZDHHC13) and ZDHHC15 expression levels were markedly increased in primary acinar cells of HLAP mice. Acinar cells of HLAP mice exhibited the increased pyroptosis, which was evidenced by the increased cleaved-caspase-1 and GSDMD-N protein levels, Interleukin (IL)-1β, IL-18, and lactate dehydrogenase (LDH) release, GSDMD-N oligomerization, and caspase-1+PI+ cells. Mechanistically, palmitic acid induced GSDMD palmitoylation. ZDHHC15 interacted with GSDMD, and ZDHHC15 knockdown reduced GSDMD-N oligomerization and thereby suppressed pyroptosis in palmitic acid-treated acinar cells. The in vivo experiments further revealed that AAV-sh-ZDHHC15 reduced cell apoptosis and GSDMD-N oligomerization in pancreatic tissues of HLAP mice. In conclusion, this study identified a potential regulatory pathway of HLAP involving ZDHHC15-dependent modulation of GSDMD palmitoylation and subsequent GSDMD-N oligomerization.
