Search Results (1,049)

Background: Autonomic side effects are a major determinant of tolerability for many psychotherapeutic drugs. While often attributed to receptor-mediated mechanisms, the potential contribution of direct modulation of smooth muscle excitability remains poorly characterized at a comparative pharmacological level. Methods: A systematic comparative pharmacological profiling of a broad panel of psychotherapeutic drugs (antidepressants, antipsychotics, and anxiolytics) was conducted using a standardized ex vivo model. Potassium chloride (KCl, 105 mM) was used to induce depolarization-dependent contraction in three isolated smooth muscle preparations (rat uterus, rat vas deferens, and guinea-pig ileum). Inhibitory potency (IC₅₀), dose-dependency, and tissue consistency were integrated to define functional inhibitory profiles. Results: Psychotherapeutic drugs exhibited marked heterogeneity in their ability to inhibit K⁺-induced smooth muscle contraction. Integrative analysis stratified compounds into four distinct functional profiles: (i) High Inhibitory Liability (e.g., nortriptyline, paroxetine), characterized by low micromolar IC₅₀ values and dose-dependent inhibition across multiple tissues; (ii) Non-Selective Inhibition (e.g., flunarizine, cinnarizine), showing consistent but dose-independent inhibition; (iii) Tissue-Dependent Inhibition (e.g., risperidone, reboxetine); and (iv) Minimal Inhibition (e.g., moclobemide). Agents classified within the High Inhibitory Liability profile correspond to drugs known to carry a higher clinical burden of autonomic adverse effects. Conclusions: This study reveals a previously underrecognized pharmacodynamic dimension of psychotherapeutic drugs and establishes a comparative functional taxonomy based on their direct, non-receptor-mediated inhibition of smooth muscle excitability. The identified profiles provide a mechanism-informed framework for contextualizing autonomic side-effect liability and may support improved safety evaluation in psychotherapeutic drug development. Full article

This experiment evaluated the effects of pellet diameter on growth performance and intestinal health of piglets during the creep feeding stage. A total of 144 7-day-old suckling piglets (body weight of 2.2 ± 0.3 kg) were randomly assigned to four groups and fed the same formula as meal feed and pellets of 2 mm, 4 mm, and 8 mm in diameter, respectively. Each treatment consisted of six replicates of six piglets. The trial was divided into two phases by weaning time: 7–21 days (breast milk + creep feed) and 21–35 days (creep feed only). After the feeding trial, piglets from the meal feed group and the 8 mm pellet group were selected for slaughter and sampling. The results showed that before weaning, average daily feed intake (ADFI) increased significantly with increasing pellet diameter (p < 0.001). Post-weaning, piglets fed 8 mm pellets presented significantly higher final body weight (FBW) and average daily gain (ADG) than those in the meal group (p < 0.05). Apparent nutrient digestibility (ATTD) in pellet groups was significantly higher than that in the meal feed group and rose with increasing pellet diameter (p < 0.001). The organ indices of the stomach and large intestine in the 8 mm group of piglets were significantly lower than those of the meal group. The jejunal villus height (VH) in the 8 mm group showed a trend toward an increase (p = 0.066), and the ileal crypt depth (CD) was significantly lower (p = 0.004), with significantly higher digestive enzyme activities in the jejunum and ileum (p < 0.05). In the 8 mm group, the relative abundances of Bacteroidetes in the jejunum and Actinobacteriota in the cecum and colon increased, while those of Pseudomonadota decreased; jejunal microbial relative richness increased significantly, while the ileal microbial operational taxonomic unit (OTU) richness decreased obviously. In conclusion, pellets improved the growth performance of creep feeding piglets. Compared with meal, 8 mm pellets can significantly enhance intestinal health level and nutrient digestion and absorption capacity by optimizing intestinal morphology, boosting digestive enzyme activities, and improving flora structure. Full article

Cerebral palsy (CP) is a non-progressive neurological condition associated with neuroinflammation, motor impairments, and gastrointestinal dysfunction mediated by the gut–brain axis. Preserving the intestinal epithelial barrier integrity may represent a therapeutic strategy, and quercetin is a bioactive compound with potential intestinal protective effects. This study investigated the effects of neonatal quercetin treatment on morphometric parameters, oxidative stress markers, and epithelial barrier gene expression in an experimental CP model. Wistar rats were distributed into four groups according to health status and treatment with a vehicle (V) or quercetin (Q, 10 mg/kg, intraperitoneally): healthy control (CV and CQ) and CP (CPV and CPQ) (n = 10/group). Intestinal morphology, oxidative stress markers, and gene expression (occludin, zonulin, and mucin 2) were evaluated. CP animals showed segment-specific alterations, with structural impairment predominantly in the ileum and increased oxidative damage in the jejunum. Quercetin attenuated oxidative stress markers and modulated antioxidant enzyme activity in CP, increased jejunal tight-junction gene expression in both healthy and CP groups, and enhanced MUC2 expression only in healthy animals, without fully reversing CP-induced morphological changes. In conclusion, neonatal quercetin modulates oxidative stress and epithelial barrier-related gene expression, supporting its potential as an adjuvant strategy for intestinal barrier protection in experimental CP. Full article

Early post-hatch feeding strategies supplementing nutrients, particularly functional amino acids, have been proposed to enhance gastrointestinal tract (GIT) maturation and health in broilers in the post-antibiotic era. However, reported effects on performance and gut morphology remain inconsistent. Hence, this meta-analysis synthesized and clarified the efficacy pattern of supplemental FAA (Arg, Gln, Gly) evidence on growth performance, gut morphology, and lymphoid organ development. From a search spanning 2015 to September 2025, data were extracted from 23 eligible studies among 582 reports identified and pooled from five online databases. Standardized mean differences (SMDs) were calculated using Hedges’ g estimator with 95% confidence intervals, and heterogeneity was explored using subgroup and meta-regression procedures. Internal validity and reliability of included studies and publication bias were also assessed. The random-effects meta-analyses revealed that the FAA increased BWG (SMD = 1.01; p = 0.0006) and reduced feed conversion ratio (SMD = −0.45; p < 0.0001). Likewise, they enhanced intestinal architecture in both the jejunum and ileum. This was characterized by increased villus height (p < 0.05), reduced crypt depth (p < 0.05), and an elevated villus-to-crypt ratio (p < 0.0001), with the ileum exhibiting the greatest morphological response. In contrast, supplementation had no significant effect on spleen weight (SMD = 0.24; p = 0.2483) or bursa weight (SMD = 0.31; p = 0.1575). These effects, however, can be influenced by dosage used, dietary crude protein level, and broiler strain. In addition to enhancing the small intestine morphology early on, longer supplementation increased feed efficiency. Specifically, L-arginine and glycine efficaciously stimulated BWG, while L-glutamine and L-arginine enhanced morphology. Overall, early dietary supplementation with arginine, glutamine, or glycine is an effective post-antibiotic nutritional strategy to alleviate early post-hatch physiological stress and support broiler growth and intestinal development. However, to optimize nutrient utilization and sustain growth performance comparable to that achieved with standard CP diets, these FAAs in practical broiler nutrition should be strategically integrated into low-CP formulations. Full article

Corn, paddy, wheat, sorghum, and cassava serve as the primary energy sources in both human and animal diets. This study aimed to evaluate their nutrient release patterns in a simulated gastrointestinal environment and to assess the in vitro biological activity of the metabolites produced during digestion. The results showed that wheat exhibited the highest dry matter degradation in the stomach–jejunum–ileum digestion stage, while wheat and paddy showed the highest crude protein degradation compared with the other starch sources. In addition, wheat had a higher total free sugar concentration than paddy, sorghum, and cassava. Among the individual free sugars, such as D-sorbitol and D-(+)-trehalose, were found to have the highest concentrations in wheat, whereas cassava had the highest D(−)-fructose concentration. Several differential metabolites, including valeric acid, caproic acid, octanoic acid, and azelaic acid were highly released in paddy, whereas glucaric acid, threonic acid, phenylacetic acid, and shikimic acid were highly released in cassava, and 4-hydroxycinnamic acid was highly released in paddy and sorghum. Four unique metabolites were identified during the digestion process of five starch sources. Particularly, isocitric acid and trans-Cinnamic acid were released only from cassava; caffeic acid was released only from sorghum and corn; and pimelic acid was released only from paddy and wheat. Furthermore, cassava was distinct from the other starch sources, displaying a higher abundance of differential metabolites within the glucagon signaling pathway as mapped in KEGG pathway analysis. In summary, compared with other starch sources, wheat provides more dry matter, protein, and sugars for the body. Cassava is unlikely to offer any advantage in glycemic regulation, while paddy and cassava possess stronger biological activity in terms of differential metabolites. Full article

Bile acids, the primary constituents of mammalian bile, are synthesized in the liver from cholesterol and secreted into the intestine to perform essential physiological functions. Primary bile acid synthesis is the principal pathway for cholesterol catabolism and whole-body cholesterol homeostasis, occurring predominantly via the classical and alternative pathways. To elucidate the effects of altitude on serum bile acid profiles and synthesis pathways in SD rats, this study utilized UPLC-MS/MS to analyze serum bile acid composition in animals housed at high and low altitudes. Additionally, qRT-PCR and Western blotting assessed mRNA transcription and protein expression of key genes involved in primary bile acid synthesis in the liver and intestinal tissues (ileum, duodenum, and colon). Results showed that serum levels of total and primary bile acids significantly decreased with increasing altitude. Furthermore, hepatic mRNA and protein expression of Cyp7a1, Cyp8b1, Cyp27a1, and Cyp7b1 were significantly downregulated. Fxr mRNA expression in the liver, ileum, duodenum, and colon was significantly decreased with increasing altitude. Meanwhile, the protein expression of both FGF15 and SHP showed a downward trend, with a significant decrease for FGF15 and a non-significant decrease for SHP. These findings suggest that primary bile acid synthesis in SD rats is dominated by the classical pathway. As altitude increases, bile acid synthesis in SD rats is significantly inhibited, indicating that high-altitude hypobaric hypoxia is the primary inhibitory factor. This study provides critical data for elucidating the adaptive mechanisms of bile acid metabolism in mammals exposed to high-altitude hypoxia, thereby establishing a theoretical foundation for investigating the regulation of host lipid metabolism influenced by such conditions. Full article

Estrogen deficiency is an established risk factor for menopausal brain dysfunctions in women. Urgent exploration of drugs is needed to improve estrogen deficiency-related brain dysfunctions without the side effects of estrogen supplements. Three-month-old rats had bilateral ovariectomy (OVX) performed and were treated with emodin (EMO, 80 mg/kg/day) and 17 β-estradiol (EST, 0.5 mg/kg/day). Brain functions were evaluated by cognition and emotion-related behavioral tests. Levels of glucagon-like peptide-1 (GLP-1) and estrogen in blood, mRNA levels of estrogen receptor (ER) α, ERβ, GLP-1 receptor (GLP-1R), proprotein convertase subtilisin/kexin type 1 (PCSK1) and proglucagon (proGCG) in intestinal segments, and brain ERα and GLP-1R levels were evaluated. Contractions of isolated intestinal segments were recorded. Additionally, an ERβ antagonist, PHTPP (200 μg/kg/day), was used to clarify the role of ERβ. EST and EMO significantly ameliorated cognition deficit and depressive behaviors in OVX rats, and reduced neuronal loss and synaptic abnormalities in the hippocampus and prefrontal cortex. The blood GLP-1 levels of sham operation rats (sham, 3.09 pg/mL), EMO-treated (2.57 pg/mL) and EST-treated OVX rats (2.64 pg/mL), were higher than that of OVX rats (1.03 pg/mL). EMO had no effect on the blood estrogen level. Furthermore, EMO up-regulated mRNA levels of ERβ in ileum, colon, and cerebral GLP-1R level, while EST increased mRNA levels of ERβ in colon and cerebral ERα level. In vitro intestinal segment spontaneous contraction tests revealed that EMO reduced contraction amplitudes in isolated intestinal segments from OVX rats, with the ileum and proximal colon showing greater sensitivity to EMO. The ileum and colon segments from OVX rats were less sensitive to EST as compared to those of normal rats. Upon PHTPP intervention, the up-regulated intestinal mRNA levels of ERβ, PCSK1, proGCG, blood GLP-1 level by EMO, and the beneficial effects of EMO in abnormal behaviors of OVX rats were significantly inhibited. Overall, it was found that EMO up-regulated blood GLP-1 level via intestinal Erβ-dependent mechanism and increased brain GLP-1R level, which may be involved in the neuroprotection of EMO in OVX animals. Full article

This study aimed to systematically evaluate the effects of low-protein (LP) diets and their associated nutritional interventions on growth performance and intestinal health in weaned piglets through a meta-analysis. Following the PRISMA guidelines, 21 studies were selected from an initial set of 1720 to construct a database. A random-effects model was employed to assess the growth performance and intestinal health of weaned piglets, comparing LP diet with control diet, and LP diet + additive with LP diet alone. Heterogeneity was assessed using the I² statistic, and its sources along with publication bias were explored through meta-regression, subgroup analysis, and Egger’s linear regression. The results show that LP diets (crude protein < 18%) reduced final body weight (p < 0.001), average daily gain (p < 0.001), and gain-to-feed ratio (p < 0.001) of weaned piglets. Additionally, forest plot data show that LP diet (crude protein < 18%) decreased villus height and crypt depth in the jejunum and ileum of weaned piglets (p < 0.05). To mitigate these adverse effects, plant extracts, amino acids (AAs), fatty acids, vitamins, enzymes, and carbohydrates are commonly used as additives. Among these, the results show that LP diets supplemented with AAs and plant extracts increased average daily feed intake (p < 0.001) and ileal crypt depth (p < 0.05). Network meta-analysis further identified AAs and plant extracts as the most effective additives for improving growth performance of weaned piglets. In conclusion, dietary protein levels below 18% negatively affect the growth performance of weaned piglets, and supplementation with AAs or plant extracts represents a promising strategy to counteract these adverse effects. Full article

The intestinal mucosal barrier is a layered structure comprising fundamental components that play important roles in regulating paracellular permeability. Disruption of intestinal barrier homeostasis predisposes to infections, mucosal damage, and metabolic and allergic diseases. To provide protection against potential damage to the intestinal mucosa, agents such as prebiotics and probiotics are recommended due to their ability to secrete components and metabolites (e.g., bacteriocins, organic acids, enzymes) that can exert beneficial biological effects. The aim of this study is to comprehensively investigate the effects of a postbiotic derived from Pediococcus acidilactici on healthy rat intestinal tissue. A total of 78 Wistar Albino rats were used in this study. Following compositional analysis of the postbiotic, the animals were administered the postbiotic orally via gavage for different durations (7, 14, 21, 28 days) and at different doses (250 mg/Kg, 500 mg/Kg, 1000 mg/Kg). Characterization of the produced postbiotic revealed a diverse spectrum of biologically active compounds, including organic acids, phenolics, and volatile compounds. Histopathological examination of intestinal sections (duodenum, jejunum, ileum, cecum, colon, and rectum) showed no pathological lesions in any of the experimental groups. Conversely, immunohistochemical analysis revealed that the postbiotic increased the expression of CLDN3, OCLN, ZO1, AQP4, and AQP8, proteins involved in intestinal permeability and fluid transport, in a dose-dependent manner. These results highlight the potential of Pediococcus acidilactici as a supportive agent in a range of intestinal pathologies, including major intestinal diseases such as Crohn’s disease, ulcerative colitis, and inflammatory bowel disease (IBD). Full article

An experiment was conducted in a commercial broiler farm to evaluate birds’ growth performance, intestinal morphology (IM), and total blood carotenoid levels when adding different inclusion rates of microbial muramidase (MUR) to broilers’ feed formula. A total of 336-day-old broilers from Ross 308 were assigned to three groups and were fed from day 1 to day 42. Treatment 1 (T1), the control treatment, was a basal corn soybean meal diet that followed Aviagen nutrient specifications; Treatment 2 (T2) was the control treatment + 350 g MUR per ton of feed; Treatment 3 (T3) was the control treatment + 450 g MUR per ton of feed. Body weight gain (BWG), feed intake (FI), the feed conversion ratio (FCR), carotenoid levels (CLs), and IM from samples of the duodenum, jejunum, and ileum were evaluated on day 42. MUR significantly outperformed the control diet in terms of body weight (BW), resulting in BWs of 2513 g (p = 0.005) and 2463 g with T3 and T2, respectively, versus 2377 g with T1. Daily weight gain (DWG) was 58.77 g and 57.58 g in T3 and T2 (p = 0.0004), respectively, versus 55.54 g in T1. The Daily Feed Intake (DFI) on day 42 was higher in T3 compared to T1 (98.90 g p < 0.0001 versus 97.72 g). The morphology results show higher effects in the duodenum with T3 versus T2 and T1. In addition, the results were significantly higher in the jejunum when using T2 and T3 compared to T1. Goblet cells were not influenced by MUR use in the diets. The broilers’ blood carotenoid levels were significantly higher in T3 (1.75 mg/L p = 0.0026) compared to T1 (1.02 mg/L). These results shed light on broilers’ performance, intestinal health, and nutrient availability when using MUR at different inclusion rates in broiler feed for broilers raised under near-commercial conditions. Full article

Dietary composition and the intestinal nutrient availability modulate metabolic pathways linked to nutrient sensors, generating molecular adaptations that influence birds’ feeding behavior and performance. The present study investigated the impact of reducing dietary crude protein (CP) and the essential amino acids (AA) lysine (Lys), methionine (Met), threonine (Thr), and tryptophan (Trp) on mRNA expression of the nutrient sensors T1R1 and T1R3, as well as on feeding behavior and productive performance in broiler chickens. A total of 64 one-day-old male Ross 308 broiler chicks were allocated to four dietary treatments: a control diet meeting recommended CP and AA levels (target live weight < 2.0 kg; T1); a diet with 3% reduction in CP and full exogenous AA supplementation (T2); a diet with 3% reduction in CP and 50% AA supplementation (T3); and a diet with 3% reduction in CP without AA supplementation (T4). After a 7-day adaptation, a 32-day single-choice test offered an AA matrix, containing Lys, Met, Thr, and Trp at eight concentrations (0.1–3.5%) for 4 h/day to assess acceptability and palatability. On day 39, six birds per treatment were sampled from the duodenum, jejunum, and ileum for quantitative PCR. Relative to controls, birds fed diets T2, T3, and T4 displayed lower feed consumption, daily gain, feed conversion ratio, and body weight (p < 0.050). These diets downregulated T1R1 and T1R3 mRNA expression (p < 0.001), indicating a molecular adaptation to the nutritional environment, and increased acceptability but reduced palatability (p < 0.050), suggesting altered feeding motivation and sensory perception. Overall, CP and AA reduction impaired performance and modified nutrient-sensor expression and feeding behavior. Full article

Background: The neutrophil-to-lymphocyte ratio (NLR) is a simple biomarker that reflects the balance between innate immune response and adaptive immunity. Currently, the genetic basis and clinical implications of NLR in relation to inflammatory gastrointestinal diseases have not been extensively explored. Methods: We carried out a genome-wide association study (GWAS) on European individuals from the UK Biobank to detect genetic variants related to NLR, followed by post-GWAS analyses including colocalization analysis, transcriptome-wide association studies (TWAS), and LD score regression. Logistic regression, Cox regression, and gene–environment interaction analysis were used to evaluate the impact of NLR polygenic risk scores (PRS) on inflammatory gastrointestinal disease risks. Results: GWAS of 395,442 Europeans identified 306 genomic regions (731 lead SNPs) associated with NLR, mapping to 1542 genes enriched for immune pathways. Colocalization revealed shared genetic signals with TWAS prioritization of 59, 19, 14, 22 and 28 genes in the whole blood, spleen, terminal ileum, transverse colon and sigmoid colon, respectively. LD-score regression showed significant positive genetic correlations with CD (rg = 0.132), coeliac disease (rg = 0.124), peptic ulcer (rg = 0.138) and duodenal ulcer (rg = 0.220). One-SD increase in NLR PRS predicted higher risk of IBD (OR = 1.05, 95% CI 1.03–1.08), Crohn’s disease (OR = 1.06, 1.02–1.10), ulcerative colitis (OR = 1.05, 1.02–1.08) and coeliac disease (OR = 1.07, 1.03–1.11). Restricted cubic splines demonstrated non-linear relationships of NLR PRS for IBD, CD and UC. Gene environment analyses showed smoking and diabetes amplified the risks, while cardioprotective diet, oily fish intake and polyunsaturated fatty acid level attenuated NLR PRS-associated risk in IBD (mainly CD). Conclusions: Our study delineates the polygenic basis of NLR and establishes its genetic correlation with inflammatory gastrointestinal diseases, offering a genetically informed indicator for disease risk stratification with potential utility in population-level prevention strategies. Full article

Background: Current artificial intelligence (AI) systems in gastrointestinal (GI) endoscopy primarily emphasize binary detection or static classification, providing limited support for the graded assessment of malignant potential that underpins clinical decision-making. We developed GastroMalign, a transformer-based framework designed to stratify GI lesions according to ordinal disease severity while maintaining clinical interpretability, addressing this unmet need in endoscopic risk assessment. Methods: This retrospective development and validation study used the publicly available GastroVision dataset, comprising 8000 de-identified endoscopic still images from the upper and lower gastrointestinal tract, including the esophagus, stomach, duodenum, colon, rectum, and terminal ileum. GastroMalign integrates a Vision Transformer (ViT) encoder with a Sequential Feature Learner that explicitly models ordinal disease severity along a benign-to-malignant spectrum. The framework produces both categorical risk classification and a continuous malignancy risk score. Images were stratified into training (80%), validation (10%), and test (10%) sets. Performance was compared with convolutional neural network (CNN) baselines and a Swin Transformer. Interpretability was assessed using Score-CAM visualizations reviewed by blinded expert endoscopists. Results: On the held-out test set (n = 800 images), GastroMalign achieved an overall accuracy of 80.06%, precision of 79.65%, recall of 80.06%, and F1-score of 79.17%, with a micro-averaged AUC of 0.98. In comparison, ResNet-50 and DenseNet-121 achieved accuracies of 32.42% and 36.77%, respectively, while the Swin Transformer achieved 60.56% accuracy (AUC = 0.93). Ablation analyses demonstrated a 17% absolute reduction in High-Risk lesion recall when the progression-aware module was removed. Continuous malignancy risk scores increased monotonically across ordinal classes, with mean values < 0.18 for Benign and >0.72 for High-Risk/Malignant lesions. Score-CAM visualizations demonstrated 92% overlap with clinician-annotated lesion regions. Conclusions: GastroMalign delivers an interpretable, progression-aware AI framework for GI lesion risk stratification that outperforms existing CNN- and transformer-based models. Clinically, GastroMalign is intended as an adjunct decision-support tool during endoscopic review to standardize lesion risk stratification (benign to malignant spectrum), support management decisions (biopsy vs. resection vs. surveillance), and reduce operator-dependent variability by pairing ordinal risk outputs with interpretable visual explanations. Full article

Background/Objectives: Enteric glial cells (EGCs) are key players in regulating enteric neurons and gastrointestinal functions including disturbed gut motility in diabetic patients. Enteric neuronal damage has been shown in type 1 diabetes, but EGCs’ vulnerability to hyperglycaemic insults requires more investigation. Therefore, we aimed to study the quantitative changes in the EGC network enmeshing enteric plexuses, intestinal smooth muscle and mucosa in streptozotocin-induced acute (1-week) and chronic (10-weeks) diabetic rat models. Methods: Fluorescent immunohistochemistry using Sox10 glial and HuC/HuD pan-neuronal markers, immunogold electron microscopy and ELISA were performed on different gut segments. Results: In the submucosal ganglia of the ileum and colon, the density of Sox10-immunoreactive EGCs was significantly reduced in acute and increased in chronic hyperglycaemic rats without any changes in the duodenum. In the myenteric ganglia, regionally distinct alterations of glial density were noted in acute hyperglycaemia; however, a remarkable decrease was observed in chronic animals. Alterations of neuronal density did not follow the pattern of glial changes, resulting in shifts in the glia/neuron ratio. The presence of Sox10-HuC/HuD-immunoreactive cells and their diabetes-related quantitative changes were also revealed in enteric plexuses. The density of Sox10-labelling gold particles was significantly increased in the duodenal myenteric glia of diabetic rats. Muscular EGC density increased only in the colon after acute hyperglycaemia and changed in all segments after chronic hyperglycaemia. Glial fibrillary acidic protein levels decreased in the small intestine of chronic hyperglycaemic rats. Conclusions: Our present findings reveal time-dependent and regionally distinct changes in the EGC network in response to hyperglycaemia, contributing to diabetic enteric neuropathy and gut motility disturbances. Full article

The present study was designed to evaluate the effects of eighteen different extracts derived from basil (Ocimum basilicum L.) leaves on spontaneous contractions, as well as contractions induced by potassium chloride (KCl) and acetylcholine in the ileum of rats, under in vitro conditions. The extracts were prepared with 96% v/v, 80% v/v, and 60% v/v ethanol, and absolute (100%) v/v, 80% v/v, and 60% v/v methanol, employing extraction techniques that included maceration, digestion, and sonication-assisted methods. Chemical characterization of the extracts revealed the presence of various phenolic acids, including rosmarinic, chlorogenic, caftaric, salvianolic acid B, cinnamic, caffeic, and chicoric acid, as well as flavonoids such as rutin and salvigenin. The evaluated extracts produced significant, concentration-dependent inhibitory effects on rat ileal contractions. Notably, the extract obtained via maceration with 80% methanol exhibited the most pronounced relaxant effects on spontaneous muscle contractions, achieving a maximum reduction of 46.16 ± 2.11%. Furthermore, the extract prepared with the same solvent using sonication-assisted extraction demonstrated superior efficacy in diminishing both the frequency and amplitude of KCl-induced ileal contractions, reducing contraction intensity caused by elevated potassium ion levels to 59.48 ± 3.34% at a maximum concentration of 1.5 mg/mL, thereby indicating its potential as a potent calcium channel blocker. Additionally, the extract prepared with 60% methanol through sonication-assisted extraction resulted in the most substantial reduction of acetylcholine-induced ileal contractions, decreasing contraction intensity to 35.74 ± 1.54% at the maximum concentration of 1.5 mg/mL, which suggests a high level of neurophysiological activity. By comparing extracts with different phytochemical profiles, this study provides additional insight into how variations in phenolic composition may influence different mechanisms of smooth muscle relaxation. This study affirms the significant spasmolytic properties of basil leaf extracts, thereby supporting their potential application in the management of gastrointestinal motility disorders. Full article