Penicillium italicum ( P. italicum ), the cause of citrus blue mould, is a pathogenic fungus that seriously affects the postharvest quality of citrus fruit and causes severe economic loss. Our previous research showed that adding limonin to eugenol nanoemulsion could improve the antifungal activity, and inhibited the growth of P. italicum on the surface of citrus fruit. Accordingly, the present study aimed to investigate the underlying mechanism of limonin-loaded eugenol emulsion (EuLm) against P. italicum based on the influence on membrane structure and intracellular substance metabolism. Results indicated that EuLm showed disruptive activity on the fungal cell membrane, mitochondrial membrane potential and the ergosterol synthesis. Meanwhile, EuLm treatment causes the intracellular substances of total lipids, cell proteins and total sugars to decrease by 56.7 %, 37.4 % and 17.4 %, respectively. GC–MS-based metabolomics showed 34 metabolites declined after EuLm treatment, including amino acids, fatty acids, carbohydrates, alcohols, and glycosides. The results suggested that EuLm interferes with fungal metabolic pathways, such as the TCA cycle, lipid metabolism and protein synthesis. Our study provides new insight into exploring the underlying mechanisms of antifungal phytochemicals against P. italicum .
PurposeMyocardial ischemia-reperfusion (I/R) is characterized by myocardial cell death and exacerbated inflammatory responses that seriously affect cardiac function and patient prognosis. Limonin is a natural compound extracted from citrus fruits and has various biological activities, including anti-inflammatory and antioxidant activities. However, the role and mechanism of action of limonin in myocardial I/R injury remain unclear. This study aimed to explore the effects and underlying mechanisms of action of limonin on myocardial cell pyroptosis and inflammatory responses in mouse myocardial I/R injury.MethodsBy analyzing the GSE225105 dataset in the Gene Expression Omnibus database, differentially expressed genes in I/R injury were screened and combined with the target genes of limonin in the Comparative Toxicogenomics Database. In the in vitro experiments, The oxygen-glucose deprivation/reoxygenation (OGD/R) model was used to simulate I/R injury in in vitro experiments.ResultsLimonin significantly inhibited OGD/R-induced pyroptosis and the release of inflammatory factors in AC16 myocardial cells, and these effects could be reversed by the caspase-3/gasdermin E (GSDME) pathway activator triclabendazole. Using in vivo experiments, we established a myocardial I/R model in C57BL/6 mice and found that limonin pretreatment improved cardiac function-related indicators, reduced myocardial tissue damage and inflammatory responses, inhibited apoptosis, and reduced myocardial fibrosis. This mechanism is closely related to the inhibition of caspase-3/GSDME pathway activation.ConclusionThis study revealed the protective effects of limonin in reducing myocardial I/R injury by inhibiting the caspase-3/GSDME pathway, thereby providing a theoretical basis for the development of limonin-based myocardial protection strategies.
Protein-based delivery systems tend to degrade too quickly in the gastrointestinal tract. Applying polysaccharide-based surface coatings on protein carriers is an effective strategy to prevent interference from gastric acid and proteases during the digestive process. This study prepared gastrointestinal pH-responsive bovine serum albumin (BSA)-myristic acid (MA) carriers using sodium alginate (SA)-based surface coating technology to improve the digestive properties and intestinal mucus penetration of limonin (LM). The results showed that ultrasonication caused uniform hydrophilic BSA and MA particle formation in optimal conditions. Fourier-transform infrared spectroscopy (FTIR) and scanning electron microscopy (SEM) observations confirmed that SA adhered to the BSA-MA particle surfaces via electrostatic interaction at pH 4, causing the controlled gastrointestinal release of the self-assembled BSA-MA complexes. The subsequent SA-BSA-MA complexes displayed an LM encapsulation yield (EY) of >84%. In vitro, simulated gastrointestinal digestion indicated that compared with free LM, the SA-BSA-MA particles significantly increased the intestinal LM retention rate, bioaccessibility, and micellization rate by 45.49%, 155.48%, and 102.14%, respectively. The results of the artificial intestinal mucus experiments demonstrated that the trans-intestinal mucus barrier capacity of SA-BSA-MA particle-loaded LM was significantly enhanced. Its apparent permeability coefficient (Papp) (6.53 × 10 −6 cm/s for free LM) and intestinal mucus permeability (17.56% for free LM) increased substantially to 14.22 × 10 −6 cm/s and 38.22%. This study presents a gastrointestinal pH-responsive delivery system that effectively improves the ability of hydrophobic nutrients to cross the intestinal mucus barrier.
危险品化学品经营许可证（带存储）