Myocardial infarction (MI) is characterized by sudden interruption of coronary blood flow, leading to ischemic damage and cardiomyocyte death. Evidence for new molecular targets remains limited. Here, we investigated the role of Schlafen 4 (Slfn4), identified via bioinformatic screening, in MI pathogenesis. We analyzed GSE46395 microarray data and observed elevated Slfn4 expression in ischemic cardiac tissue. An MI mouse model further confirmed Slfn4 upregulation, which was abrogated by AAV9-mediated shRNA knockdown. Silencing Slfn4 reduced inflammatory cell infiltration and cardiomyocyte apoptosis, leading to lower serum levels of ANP, BNP, cTnT, cTnI, IL-1β, and TNF-α. Notably, Slfn4 knockdown augmented BNIP3-dependent mitophagy, evidenced by upregulated LC3 I/II, decreased P62, and reduced mitochondrial proteins (COX IV, TOMM20), while also suppressing DRP1-mediated mitochondrial fission. In cultured H9C2 cells subjected to hypoxia, Slfn4 knockdown likewise diminished apoptosis and enhanced BNIP3-associated mitophagy, whereas BNIP3 silencing reversed these protective effects, underscoring the importance of BNIP3-mediated mitophagy in Slfn4-driven cardioprotection. These findings indicate that Slfn4 promotes MI-induced damage by inhibiting BNIP3-mediated mitophagy and exacerbating mitochondrial fission. By contrast, Slfn4 knockdown fosters cardiomyocyte survival, highlighting its therapeutic potential for MI. Overall, our data suggest that modulating Slfn4 expression may preserve mitochondrial quality control, attenuate inflammation and apoptosis, and improve cardiac function following ischemic injury. .

We have investigated the involvement of K channels in generating the membrane current in MDA-MB-436 cells, a model of triple-negative breast cancer (TNBC). The membrane current is strongly influenced by the opening of voltage-dependent channels insensitive to the nonspecific K channel inhibitor 4-aminopyridine (4-AP). Using the cell patch clamp technique, we observed a significant decrease in membrane current after exposure to the generic K channel inhibitor tetraethylammonium chloride (TEA-Cl), indicating that K ions contribute to the overall membrane current through K channels that are insensitive to 4-AP but TEA-Cl-sensitive. RNA-sequencing analysis identified the Big Potassium (BK or Maxi-K or KCa1.1, encoded by KCNMA1) and the Kv2.1 (encoded by KCNB1) channels as putative candidates, both of which are involved in cancer cell proliferation and migration. Iberiotoxin, a specific inhibitor of BK channels, did not affect the total membrane current, just as CdCl₂ did, a potent inhibitor of Ca⁺ channels involved in BK activation. Using selective inhibitors, stromatoxin and drofenine, we demonstrated that the Kv2.1 channel contributes to the membrane current in MDA-MB-436 cells. Furthermore, drofenine inhibited cell migration as measured by the xCELLigence Real-Time Cell Analyzer System and induced apoptosis. Single-cell analysis revealed that the Kv2.1 channel is expressed in both normal and cancerous tissues, with significant upregulation in brain metastases. This raises the possibility that Kv2.1 could be explored as a potential therapeutic target for controlling advanced stages of the neoplasia.

Age-associated sarcopenia is characterized by progressive loss of skeletal muscle mass and function. Irisin, a myokine, has been shown to improve sarcopenia; however, the dosage-dependence of its effects and the underlying molecular mechanisms remain unclear. To investigate the effects of irisin on age-associated sarcopenia, 22-week-old mice were used. Recombinant irisin was administered via intraperitoneal injections at doses of 0.5, 1, and 2 mg/kg, three times per week, to evaluate potential dosage-dependent effects. Skeletal muscle function was assessed using hanging time, grip strength, and muscle mass measurements. Morphological changes in muscle tissue were examined through hematoxylin and eosin staining, and fibrosis was quantified using Masson staining. Serum irisin levels were measured via enzyme-linked immunosorbent assay, and protein expression was analyzed using Western blotting. Recombinant irisin treatment significantly increased serum irisin levels in aged mice and improved functional metrics, including hanging time, maximum speed, grip strength, and muscle mass, in a dosage-dependent manner. Histological analysis revealed improvements in muscle structure and a reduction in fibrosis following irisin treatment. Molecular analyses suggested that irisin may modulate iron homeostasis and restore key oxidative stress-related proteins such as GPX4 and SLC7A11. Further exploration revealed that irisin treatment restored sirtuin 1 (SIRT1) levels, leading to deacetylation of P53 and subsequent reduction in its expression. Irisin treatment ameliorates age-associated sarcopenia in a dosage-dependent manner, potentially involving iron overload and the SIRT1/P53 pathway. These findings provide insights into the therapeutic potential of irisin for age-related skeletal muscle atrophy.

High-altitude hypoxia (HH) significantly affects mammalian physiological functions, especially sleep rhythms, anxiety-like behavior, and neuroinflammation. In this study, adult male Sprague-Dawley rats were randomly assigned to five groups: Control, HH, HH with low-dose dexmedetomidine (Dex), HH with high-dose Dex, and Control with Dex. Rats were exposed to a simulated 6000-meter altitude for 7 days. Behavioral tests, enzyme-linked immunosorbent assay (ELISA), reverse transcription quantitative PCR (RT-qPCR), and Western blotting were used to assess hormone levels, gene/protein expression, and inflammatory markers. HH exposure elevated hypothalamic serotonin (5-hydroxytryptamine, 5-HT) and reduced melatonin levels. The sleep rhythm-related gene Timeless was downregulated, while Homer1 was upregulated at both mRNA and protein levels. Knockdown of Timeless or overexpression of Homer1 resulted in a significant increase in hypothalamic 5-HT levels and a marked decrease in melatonin levels. HH induced enhanced anxiety-like behaviors and reduced spontaneous activity, accompanied by elevated levels of L-1β, IL-6, and TNF-α in both serum and hypothalamus. Inhibition of the TLR4-MyD88-NFκB pathway significantly alleviated both hormonal disturbances and anxiety-like behaviors in HH rats.Dex treatment, especially at high doses, suppressed inflammatory responses, normalized hormone levels, restored sleep gene expression, and improved behavioral outcomes. These results indicate that dex mitigates HH-induced sleep and inflammatory disturbances, highlighting its therapeutic potential for high-altitude neurological dysfunction.

Previous studies have indicated that miR-592 may play a significant role in the development of various cancers, particularly in the proliferation and growth of breast cancer (BC) cells. However, the regulatory mechanism by which miR-592 influences apoptosis in BC cells remains unclear. In this study, functional assays investigating the role of miR-592 in apoptosis of MCF-7 cells were conducted, alongside transcriptome analysis using RNA-seq. The results demonstrate that miR-592 not only inhibits breast cancer progression but also promotes apoptosis in breast cancer cells. Several significantly dysregulated genes and enriched pathways were identified in response to miR-592 overexpression in MCF-7 cells. Among these, CYP20A1 was validated as a key target gene of miR-592, with its expression being negatively regulated by miR-592. Restoration of CYP20A1 expression reversed the pro-apoptotic and tumor-suppressive effects of miR-592 in MCF-7 cells. This process was mediated through activation of the C/EBP homologous protein (CHOP)-dependent endoplasmic reticulum stress-induced apoptotic pathway. Collectively, these findings indicate that CYP20A1 is involved in the regulation of the CHOP-mediated endoplasmic reticulum apoptosis pathway in miR-592-induced apoptosis in breast cancer cells.

Ferroptosis is a kind of programmed cell death characterized by the iron-dependent lipid peroxides accumulation, playing a pivotal role in the pathogenesis of various diseases, including neurodegenerative disorders, cardiovascular diseases, and osteoporosis. Mesenchymal stem cells (MSCs) and MSCs-derived exosomes (MSC-exos) are actively implicated in key biological processes, such as inflammatory and immune responses, tissue regeneration and repair, and aging. Emerging studies highlight the potential of MSCs and MSC-exos as effective regulators of ferroptosis, offering novel strategies for targeted therapeutic intervention in ferroptosis-related pathologies. This review comprehensively explores the precise regulatory mechanisms by which MSCs and MSC-exos modulate ferroptosis. We also evaluate the impact of ferroptosis on MSC biological functions and MSC-exos release. Furthermore, the therapeutic potentials and advantages of engineered MSCs and MSC-exos in the treatment of various diseases have also been explored, emphasizing their mechanistic roles in ferroptosis modulation across different organs and systems. This review provides insights and future directions for the development of novel MSC- or MSC-exos-based therapeutic strategies targeting ferroptosis.

Addressing the physiological effects of bioactive compounds in metabolic diseases (i.e., obesity, diabetes, liver steatosis) and establishing their mechanisms of action have been a major interest for the last decades. However, methodologies that can be applied to achieve this can vary greatly, leading to a limited type of information. Thus, the accuracy, robustness, reliability and potential (human) translation are highly reliant on the experimental design and selected methodological models. This review presents an update exploring the main features, advantages and disadvantages of most important pre-clinical models used at the present time to study the effects of bioactive compounds on metabolic diseases. Moreover, future challenges in developing new methods are also depicted. In vitro models (enzyme assays and standard two-dimensional cultures of adipocytes, skeletal muscle cells) are intrinsically well established and constitute the first choice and most widely used methods to study bioactive compounds in metabolic diseases. However, novel models such as three-dimensional cultures (spheroids, organoids) are also starting to emerge and complement traditional culture systems. Models of small organisms (C. elegans, D. melanogaster) and non-mammal vertebrates (D. rerio) represent a scientific advantage and a middle-step before traditional mammalian models (rats and mice). This article provides extensive information and a critical overview of a wide range of methods that represent present and future avenues towards a further understanding of metabolic diseases. Combining and developing new methods will be key for future progression on the effects of bioactive compounds on metabolic diseases, as well as to minimize the use of mammalian models due to ethical reasons.

Interscapular brown adipose tissue (iBAT) whitening is characterized by thermogenic dysfunction and may aggravate the metabolic complications of obesity. Moreover, whitening is linked to multiple factors, including the overactivation of the classical renin-angiotensin system (RAS) arm. Thus, we aim to compare the impact of RAS modulation by enalapril and/or aerobic exercise training (AET) in obesity-induced iBAT whitening. C57BL/6 mice were fed a standard chow (SC) or high-fat (HF) diet for 16 weeks. After 8 weeks, the HF animals were subdivided (n = 10 for each group): HF, HF + Enalapril (HF-E), HF + Training (HF-T), and HF + Enalapril + Training (HF-ET). We evaluated: body mass (BM) gain, adiposity index, iBAT morphology, RAS components, and thermogenic markers in iBAT. The HF group exhibited increased body adiposity, iBAT mass, and lipid content, as evidenced by a collapse in iBAT thermogenic capacity and overactivation of the classical RAS arm (AT1R) compared to the SC group. All interventions reduced body adiposity and iBAT mass, improved thermogenic capacity (UCP1, Vegfa, and AMPK), and shifted iBAT RAS balance to the counter-regulatory arm (MasR and MrgD). HF-E improved iBAT morphology and brown adipogenesis (Prdm16 and Cidea) compared to the HF group. HF-T improved mitochondrial biogenesis (PGC1α and Tfam) compared to the HF group. HF-ET showed additional reduced body adiposity by sympathetic activation (β3-ar) and increased brown adipocyte competence (AMPK, Tfam, and Cidea) compared to SC and HF groups. We concluded that combined interventions (enalapril and AET) have an additional impact on iBAT whitening, enhancing brown morphology and thermogenic capacity in obese mice.

Aerobic training (beneficial for type 1 diabetes mellitus (T1DM) patients) could result boring, leading to inactivity or suboptimal performance. Therefore, High-Intensity Interval Training (HIIT) could be an appealing alternative. The present study aimed to compare modulation of certain inflammatory cytokines in T1DM participants performing aerobic vs. HIIT routines. We recruited 26 T1DM male subjects: ages 18-40, T1DM ≥ 2 years, glycated hemoglobin < 8.5%, stable insulin regimen for 6 months, minimum 90 min weekly physical activity and completion of International Physical Activity Questionnaire (IPAQ). IPAQ:2 corresponds to moderate activity and IPAQ:3 to intense physical activity. Participants performed a single aerobic or HIIT session separated by at least 72 h. Blood plasma samples were collected 20 min before and after each session. Cytokines were measured using LUMINEX technique. After aerobic exercise, pro-inflammatory cytokines interleukin (IL)-1β, IL-6, IL-7, IL-8, IL-17 A, tumor necrosis factor-α (TNF-α) and interferon-γ (IFN-γ) showed no significant differences in participants (IPAQ:2 and 3). Only anti-inflammatory cytokine IL-2 increased significantly in IPAQ:3 participants. Compared to pre-exercise, post-exercise HIIT situation presented a similar pattern. The 4 participants (IPAQ:3) that voluntarily followed a 12-week HIIT routine, showed significant increases in IL-7, IL-8, and TNF- α, and the detection of the anti-inflammatory cytokine IL-22. Altogether, these results suggest that HIIT favors the presence of some pro-inflammatory cytokines. The anti-inflammatory action of certain cytokines, such as IL-22, should be considered for a possible compensatory action. Nevertheless, programs of interval exercises at moderate intensity could be at the moment a safe option for T1DM patients.

Food allergy (FA) is an exacerbated immune system response to harmless food antigens following sensitization. The incidence of FA has risen significantly over the past two decades, a trend often attributed to modern lifestyle factors such as dietary patterns, antibiotic use, and urban environments. Sensitization may result from a compromised intestinal barrier caused by inflammatory bowel diseases, genetic predisposition, or a combination of both. These conditions trigger an inflammatory response involving mechanisms such as the activation of Toll-Like Receptors (TLRs), which recognize pathogen-associated molecular patterns. This review examines the intestine's role as a key antigen-sensing organ through three critical components: a) gut-associated lymphoid tissue, b) the mucosal immune system, and, c) the intestinal microbiota in the development of FA. The role of TLRs (particularly TLR2 and TLR4) in recognizing bacterial membrane-derived compounds (e.g., lipopolysaccharides) and how commensal bacteria generate TLR ligands that influence allergen sensitization vs. tolerance is discussed. The importance of candidate gene polymorphisms encoding TLR proteins and other molecules associated with tolerance and sensitization to food antigens is also commented on. Finally, future research directions and preventive strategies to mitigate FA risk and development are suggested.

Cold injury presents a significant health challenge, causing tissue damage due to prolonged exposure to low temperatures. This study examines menthol's protective effects against cold injury, focusing on its activation of transient receptor potential cation channel subfamily M member 8 (TRPM8), a "cold-sensing" receptor, to stimulate thermogenesis in brown adipose tissue (BAT). Male C57BL/6J mice were treated with menthol for 21 days and exposed to -20 °C. Core body temperature, activity levels, and cold injury severity were measured. Network pharmacology methods identified TRPM8 as a potential target, confirmed through molecular docking and pathway analysis. Further experiments inhibited TRPM8 to evaluate its role in menthol-induced thermogenesis and cold tolerance. Menthol significantly raised core body temperature, improved cold tolerance, and reduced cold injury severity in treated mice. Network pharmacology analysis highlighted TRPM8 as a key regulator of BAT thermogenesis through the PKA/UCP1 pathway. TRPM8 inhibition diminished menthol's effect, underscoring its essential role in menthol-mediated thermogenesis. This study demonstrates that menthol activates TRPM8 in BAT, enhancing thermogenesis to prevent cold injury. These findings suggest menthol as a promising natural agent for cold injury prevention, with TRPM8 as a potential therapeutic target.

Mediated by the bitter taste receptors (TAS2R), the perception of bitter taste does not only involve the oral cavity but various physiological systems throughout the gastrointestinal tract. The relationship between stimulation and modulation is crucial for understanding the broader implications of bitter taste signalling in health and disease. In this study, we investigated how the expression of intestinal rat Tas2r (rTas2r) is affected by natural extracts containing bitter ligands, examined their association with obesity, and their effects on GLP-1 secretion. For this, we performed subchronic stimulations with a mixture of polyphenols and individual molecules in rats. Moreover, we also examined how the individual bitter molecule (epicatechin) affects the secretory profile of intestinal enteroendocrine cells. Treating rats with procyanidins up-regulated rTas2r in all the segments of the gastrointestinal tract, with the most changes observed in the duodenum and ascending colon. Epicatechin, one of the main components of the previously used extract, had a much more specific effect, as we observed mostly changes in the jejunum, where rTas2137, -139, -143 and -144 were up-regulated. In Hutu-80 cells, epicatechin downregulated TAS2R14 after 24 hours, which limited GLP-1 secretion after acute peptone stimulation. Our results support a network effect in the role of the bitter taste receptors along the intestinal areas that must be considered to address the work with bitter agonists.

Acute intermittent porphyria (AIP) is a genetic metabolic disorder characterized by neurovisceral attacks. Although high-carbohydrate diets or intravenous glucose administration can help alleviate incipient attacks in patients, these interventions may also promote insulin resistance and increase metabolic risk. This study explored targeted dietary interventions to manage hyperinsulinemia and to enhance glucose uptake in insulin-sensitive organs under high-carbohydrate diet. Body composition and fecal microbiota profile were also investigated in a murine model of the disease. Wild-type and AIP mice (n = 6/group) were supplemented with tapioca maltodextrin in drinking water for 12 weeks, alongside heat-treated Bifidobacterium animalis subsp. lactis CECT-8145 (BPL1®HT), its by-product lipoteichoic acid (LTA), or the insulin-sensitizing agent α-lipoic acid (α-LA). Liver-targeted therapies, previously assessed in AIP mice, were also included in this study. AIP mice on a high-carbohydrate diet exhibited hyperinsulinemia and tissue-specific differences in glucose uptake compared to wild-type mice. Dysbiosis, marked by reduced fecal Dorea spp. and Adlercreutzia muris, alongside higher abundance of Escherichia coli, was also showed. Supplementation with α-LA and LTA revealed superior ability to improve glucose tolerance test and skeletal muscle glucose uptake, reduce hyperinsulinemia, and enhance body composition by increasing lean mass relative to fat, compared to gene therapy or liver-targeted insulin administration. Notably, LTA restored fecal microbiota profiles resembling those of wild-type mice. In conclusion, supplementation with LTA from BPL1®HT and α-LA may represent promising dietary interventions to manage glucose tolerance, improve insulin sensitivity in muscle and adipose tissues, and potentially ameliorate body composition in AIP patients under a high-carbohydrate diet.

This study aimed at determining the effects of type II collagen (CII)-induced arthritis (CIA) on cardiac homeostasis in the contexts of a laboratory chow (LC) and a Western diet (WD). The influence of dietary docosahexaenoic acid (DHA) was also examined. Sixty female Wistar rats were assigned to five groups. The first two groups were fed the LC and were treated or not with CII (LC + CIA and LC); the third and fourth groups were fed a WD with or without CII treatment (WD + CIA and WD); and the fifth group was treated with CII and it was fed the WD whose 2.5% of the lipid fraction was replaced by DHA (DHA + CIA). Ionic homeostasis, redox status, inflammation markers, and mitochondrial stress were analysed in the heart. CIA reduced the body weight and favoured wasting of the lipid and protein stores. It also reduced cardiac cell density. The CIA subgroups, particularly the WD + CIA rats, showed higher cardiac calcium and lower reduced glutathione to oxidized glutathione ratio. In the LC + CIA rats, no oxidative/nitrosative stress (ONS) was noticed and the mitochondrial extraction yield (MEY) was similar to that measured in the LC subgroup. In contrast, the ONS was higher and the MEY was lower in the WD + CIA subgroup compared to the WD one. The observed differences were not due to inflammation. DHA had little effect on the cardiac consequences of CIA. In conclusion, the WD amplified the deleterious effects of CIA on cardiac homeostasis by weakening the mitochondria via an increased ONS.

Malnutrition of protein and essential nutrients in children can lead to serious health problems. It significantly alters hepatic physiology and leads to impaired liver function. The present study investigated the underlying mechanism of malnutrition-induced steatohepatitis in a rat model. Weanling rats were divided into two groups. The control rats received a standard protein diet, while the other group was fed a low protein diet (LPD) for eight weeks. LPD significantly reduced the body and liver weights and altered the blood parameters. LPD resulted in elevated serum liver injury markers and lowered glucose, albumin, and total protein levels. The reduced levels of TIBC and TSI and upregulated expression of Hamp gene were observed in the LPD group. Histopathology revealed the severe fat accumulation in the hepatocytes, leading to inflammation and fibrognesis. LPD upregulated the de novo lipogenesis (Srebp1c, Fas, Acc, and Scd1) markers and oxidative stress in the hepatic tissue. The downregulation of Pgc1α, Tim23, and Tfam indicated mitochondrial dysfunction in the LPD group. Transcriptomic analysis revealed the upregulation of 7,545 genes in the LPD group mainly associated with metabolic dysfunction-associated steatotic liver disease (MASLD), beta-oxidation, AMPK signalling and oxidative phosphorylation. Hepatic lipidome revealed the elevated levels of various lipid species in the LPD group. Further, LPD altered the gut microbiome of rats and reduced the relative abundance of beneficial bacteria. The present study revealed that malnutrition induces hepatic steatoheptitis by altering the hepatic lipid metabolism and disrupting mitochondrial function and gut-liver axis.

The farnesoid X receptor (FXR), a nuclear receptor (NR), plays a key role in balancing bile acid (BA), lipid, and glucose metabolism. By partnering with the retinoid X receptor (RXR), FXR influences gene transcription critical to these metabolic pathways. It also interacts with other NRs, including the pregnane X receptor (PXR), liver X receptor (LXR), and vitamin D receptor (VDR), creating an intricate signalling network. FXR activation triggers the production of small heterodimer partner (SHP), which suppresses cholesterol 7 alpha-hydroxylase (CYP7A1), the enzyme controlling BA synthesis. It also regulates lipid metabolism by controlling sterol regulatory element-binding protein 1c (SREBP-1c) and affects glucose balance. LXR, activated by oxysterols, supports reverse cholesterol transport (RCT) by regulating the expression of adenosine triphosphate binding cassette A1 (ABCA1) and adenosine-binding cassette sub-family G member 1 (ABCG1). Since FXR affects LXR-regulated genes, it indirectly modulates cholesterol homeostasis. Meanwhile, PXR, a xenobiotic sensor responsive to diverse compounds, such as BAs, regulates genes involved in drug detoxification and transport. FXR activation enhances PXR expression, influencing BA metabolism and removal. VDR, which responds to vitamin D and specific BAs such as lithocholic acid, plays a role in calcium balance and xenobiotic processing. The interplay among these NRs underscores FXR's central role in metabolic regulation and its potential as a therapeutic target for metabolic disorders.

Long non-coding RNAs (lncRNAs), defined as transcripts exceeding 200 nucleotides without protein-coding potential, have emerged as pivotal regulators in diverse physiological and pathological processes, particularly in tumorigenesis. Among them, NR2F1-AS1, a recently characterized lncRNA, has garnered growing attention due to its dysregulated expression across a spectrum of malignancies and its significant correlation with key clinicopathological parameters. Accumulating evidence from molecular and cellular studies reveals that NR2F1-AS1 plays multifaceted roles in cancer initiation and progression through the modulation of signaling pathways, regulation of gene expression, and interactions with microRNAs and protein complexes. Notably, its biological function appears to be context-dependent: acting as an oncogene in many cancer types, such as breast, lung, liver, and gastric cancer, while exhibiting potential tumor-suppressive activity in others, including colorectal cancer, cervical squamous cell carcinoma, and thymic epithelial tumors. This review comprehensively summarizes the aberrant expression patterns, prognostic significance, biological functions, and molecular mechanisms of NR2F1-AS1, while also highlighting its emerging potential as a context-specific diagnostic biomarker and therapeutic target in human cancers.

Ceramides are sphingolipids formed from fatty acids linked to sphingosine and an amide, which are involved in cellular pathways such as apoptosis, fibrosis, oxidative stress, and inflammation. Six distinct fatty acyl selective ceramide synthases (CerS) produce ceramides. This specific enzymatic modulation can either increase or reduce the production of specific ceramides, which can have either adverse or protective effects, suggesting that enzymatic modulation may serve as a tool for innovative therapy. Specifically, modulation of glucosylceramide synthase, sphingomyelinase, or ceramidase can reverse the generation of potentially apoptotic ceramides, similar to how inhibition of serine palmitoyltransferase or ceramide synthases may be significant in inflammatory conditions by decreasing the generation of inflammatory ceramides. In this context, the modulation of plasma ceramides may represent a protective factor for chronic non-communicable diseases (NCDs), such as cardiovascular diseases, type 2 diabetes, and chronic kidney disease. Previous studies indicate that dietary fat and protein intake influence plasma sphingolipid levels. Therefore, this review aims to discuss the effects of ceramide on patients with NCDs, providing an overview of the influence of nutrition on ceramide levels and outlining future perspectives.