BACKGROUND: Synthetic food preservatives are widely used to improve food stability and shelf life, but prolonged exposure may contribute to oral and gastrointestinal carcinogenesis. Epidermal growth factor receptor (EGFR)–phosphoinositide 3-kinase (PI3K)–protein kinase B (Akt)–mammalian target of rapamycin (mTOR) axis plays a pivotal role in regulating tumor initiation and progression; however, the potential interactions with cancer-related signaling proteins remain poorly understood. This study computationally evaluated the predicted binding interactions of commonly used synthetic food preservatives with proteins using molecular docking. METHODS: Propyl gallate (PG), butylated hydroxyanisole (BHA), tertiary-butylhydroquinone (TBHQ), potassium sorbate (PS), sodium benzoate (SB), and sodium metabisulfite (SMB) were evaluated for physicochemical properties using SwissADME. Molecular docking with EGFR, PI3K, Akt-1, and mTOR was performed using CB-Dock 2.0. Predicted ligand–protein interactions were analyzed using BIOVIA Discovery Studio 2016 and root mean square fluctuation (RMSF) was evaluated using CABS-flex 3.0. RESULTS: PG and BHA exhibited the most predicted binding affinities among the investigated preservatives. PG showed the strongest interactions with EGFR (−6.1 kcal/mol), PI3K (−6.8 kcal/mol), and mTOR (−6.7 kcal/mol), whereas BHA demonstrated the highest affinity toward PI3K (−6.8 kcal/mol) and Akt-1 (−6.5 kcal/mol). These interactions were supported by multiple hydrogen bonds, hydrophobic interactions, and van der Waals contacts. CONCLUSION: PG and BHA exhibited the highest binding affinity toward proteins of the EGFR–PI3K–Akt–mTOR associated with oral and gastrointestinal carcinogenesis. These computational findings provide a basis for future experimental studies to determine the biological relevance of the predicted protein–ligand interactions under long-term dietary exposure. KEYWORDS: synthetic food preservatives, molecular docking, oral carcinogenesis, gastrointestinal carcinogenesis
BACKGROUND: Current management of traumatic brain injury (TBI) is primarily directed toward preventing secondary brain injury, while specific pharmacological therapies targeting post-traumatic neuroinflammation remain limited. Bioactive constituents of Nigella sativa, particularly thymoquinone, have demonstrated anti-inflammatory and antioxidant properties that may modulate mechanisms involved in secondary brain injury. This study was conducted to evaluate the effects of oral N. sativa oil on serum interleukin (IL)-1β and IL-10 levels as inflammatory parameters, neurological function and necrosis in a rat model of TBI. METHODS: Sixteen male Wistar rats underwent Marmarou weight-drop TBI and were randomly assigned into two groups given either oral N. sativa oil or distilled water (n=8/group) for 7 days. Serum IL-1β and IL-10 concentrations were measured using rat-specific enzyme-linked immunosorbent assays (ELISA), and neurological function was assessed using the modified Neurological Severity Score (mNSS) at post-injury baseline and on treatment day-1, -3, and -7. After day-7, brain tissue was examined using hematoxylin–eosin (H&E) staining, and necrotic-cell count was assessed. RESULTS: IL-1β concentrations were significantly lower in the N. sativa group from day-1 onward, whereas IL-10 concentrations were significantly higher and mNSS scores were significantly lower from day-3 onward. By day-3, all three outcomes differed significantly: lower IL-1β (56.38±10.21 vs. 111.98±47.64 pg/mL; p=0.013), higher IL-10 (102.38±33.42 vs. 40.20±6.02 pg/mL; p=0.001), and lower mNSS (median: 4 (3-5) vs. 5 (5-6); p=0.003) than placebo group. These differences persisted through day-7, which also supported by lower necrotic-cell counts in the N. sativa group (3164.13±771.38 vs. 4028.38±204.58; p=0.008). CONCLUSION: Administration of oral N. sativa oil shifted the circulating cytokine profile toward an anti-inflammatory pattern and was accompanied by better neurological function and fewer neuronal necrosis after TBI. These findings suggest that N. sativa oil may have beneficial effect on neuroinflammation after TBI. KEYWORDS: Nigella sativa, traumatic brain injury, interleukin-1 beta, interleukin-10, neuroinflammation, modified Neurological Severity Score
BACKGROUND: Chronic inflammation contributes to diabetes-associated fibrosis, through persistent hyperglycemia, leading to increased interleukin-6 (IL-6), transforming growth factor-β (TGF-β), and collagen levels. While metformin effectively controls blood glucose, it may not adequately suppress these pathogenic pathways. Asiatic acid has demonstrated anti-inflammatory and antifibrotic properties, although its effects within a direct-contact macrophage–fibroblast microenvironment such as in co-culture models remain unclear. This study was conducted to evaluate the effects of asiatic acid on Il6, Tgfb1, and soluble collagen levels in a RAW 264.7–NIH 3T3 co-culture under high-glucose and lipopolysaccharide (HG–LPS) stimulation. METHODS: RAW 264.7 and NIH 3T3 cells were co-cultured at ratios of 1:1–1:6, and the selected co-culture was treated with asiatic acid (2.5, 5.0, and 10.0 µg/mL) for 24 h while HG–LPS stimulation was maintained. Unstimulated NIH 3T3 monocultures and untreated HG–LPS co-cultures served as controls. The inflammatory and profibrotic markers such as Il6 and Tgfb1 mRNA expression were quantified by polymerase chain reaction (PCR), and soluble collagen was measured using a modified Sirius Red assay. RESULTS: Among the tested ratios, the 1:1 co-culture exhibited the highest Tgfb1 mRNA expression (0.35±0.02) and soluble collagen level (1.30±0.00) and was selected for subsequent experiments, whereas Il6 mRNA expression did not differ significantly among co-culture ratios. Under HG–LPS stimulation, Il6 and Tgfb1 mRNA expression increased 3.5-fold and 10.3-fold, respectively, compared with unstimulated NIH 3T3 monocultures. Asiatic acid significantly reduced Il6 mRNA expression at 5.0 and 10.0 µg/mL, Tgfb1 mRNA expression at 2.5 µg/mL, and soluble collagen levels at all tested concentrations, indicating a non-monotonic concentration–response pattern. CONCLUSION: The 1:1 RAW 264.7–NIH 3T3 co-culture was the most responsive under HG–LPS stimulation and could be in vitro platform for investigating macrophage–fibroblast interactions under hyperglycemic inflammatory conditions. Asiatic acid modulated Il6 and Tgfb1 mRNA expression and soluble collagen levels in RAW 264.7–NIH 3T3 co-culture, which suggest that Asiatic acid may be potential for the management of fibrotic inflammation. KEYWORDS: co-culture, collagen, fibroblast dysfunction, NIH 3T3 fibroblast, RAW 264.7 macrophage
BACKGROUND: Cyclea barbata Miers has been identified as a phytoestrogenic candidate with comparable estrogen receptors (ER)α binding activity with 17β estradiol. Though phytoestrogens are reported to modulate reproductive and hormonal physiology, but no systematic in vivo toxicological evaluation of C. barbata extract has been reported. Therefore, this study was conducted to evaluate the acute and subacute oral toxicity of the ethanol extract of C. barbata (EECB) in Wistar rats. METHODS: For acute toxicity, rats received single oral doses of 2,000 or 5,000 mg/kg BW and were observed for 14 days. For subacute toxicity, EECB was administered orally at doses of 100–500 mg/kg BW/day for 28 consecutive days. Body weight, relative organ weights, haematological and biochemical parameters, and histopathological changes in major organs were evaluated. RESULTS: No mortality occurred at 2,000 mg/kg BW in either sex, whereas mortality was observed only in female rats at 5,000 mg/kg BW (20%). EECB was therefore classified as GHS Category 5 or unclassified (LD50 >2,000 mg/kg BW), indicating low acute oral toxicity. Repeated administration for 28 days caused no significant changes in body weight, organ weights, serum biochemical markers, or histopathological findings. However, a significant reduction in white blood cell counts was observed at doses ≥400 mg/kg BW/day (p<0.05). The subacute no-observed-adverse-effect-level (NOAEL) and lowest-observed-adverse-effect-level (LOAEL) were established at 300 and 400 mg/kg BW/day, respectively. CONCLUSION: Based on the parameters assessed, the acute NOAEL for EECB was 2,000 mg/kg BW, while the subacute NOAEL and LOAEL for EECB were 300 and 400 mg/kg BW/day, respectively. These findings indicate that EECB might be safe at moderate doses, providing in vivo evidence supporting its further development as a phytoestrogenic agent. KEYWORDS: Cyclea barbata Miers, ethanol extract, acute toxicity, subacute toxicity, haematological parameters
BACKGROUND: Diabetes mellitus often exacerbated by cigarette smoke, which disrupts glucose homeostasis and lipid metabolism. Mango (Mangifera indica L.) and lime (Citrus amblycarpa) peels contain various phytochemical compounds, including those are reported to have metabolic and cytoprotective properties, but their combined nanoparticle formulation in cigarette smoke-exposed diabetes cases remains unexplored. Therefore, this study was conducted to evaluate the synergistic effect of mango-lime peel nanoparticles (NanoMC) on glycemic control, lipid profiles, and pancreatic cell damage in cigarette smoke-exposed diabetic rats. METHODS: Mango and lime peel ethanol extract was prepared into nanoparticles using emulsion-solvent evaporation method and followed by sonication and homogenization, and subsequently divided into three concentration groups. Rats were diabetic-induced and smoke-exposed, before being treated with/without Simvastatin + Glibenclamide, 20, 40, or 80 mg/kgBW NanoMC. Parameters including blood glucose, insulin, homeostatic model assessment of insulin resistance (HOMA-IR), homeostatic model assessment of beta-cell function (HOMA-β), hemoglobin A1c (HbA1c), and lipid profiles were then measured. To evaluate pancreatic cell damage, the pancreatic tissue was collected, fixed, and then evaluated by using Hematoxylin and Eosin (H&E) staining. RESULTS: NanoMC administration significantly reduced blood glucose, HbA1c, triglycerides, cholesterol, and low-density lipoprotein (LDL) levels (p<0.05). NanoMC treatment also increased insulin levels, HOMA-β, and high-density lipoprotein (HDL). After the treatment with NanoMC, particularly with 80 mg/kgBW concentration, fewer degenerative and necrotic cells were found compared to the other groups, showing its ability to attenuate pancreatic cell damage. CONCLUSION: The combination of NanoMC could improve glycemic status, modulate lipid profiles, and improved pancreatic cell damage in diabetic rats exposed to cigarette smoke. KEYWORDS: Mangifera indica, Citrus amblycarpa, nanoparticles, diabetes mellitus, cigarette smoke, lipid profiles, pancreatic histopathology
