Glycolysis can improve the tolerance of tissue cells to hypoxia, and its intermediates provide raw materials for the synthesis and metabolism of the tumor cells. If it can inhibit the activity of glycolysis-related enzymes and control the energy metabolism of tumor, it can be targeted for the treatment of malignant tumor. The target proteins phosphoglycerate kinase 2 (PGK2), glycerol-3-phosphate dehydrogenase (GPD2), and glucose-6-phosphate isomerase (GPI) were screened by combining transcriptome, proteomics, and reverse docking. We detected the binding constant of the active compound using microscale thermophoresis (MST). It was found that esculetin bound well with three potential target proteins. Esculetin significantly inhibited the rate of glycolysis, manifested by differences of cellular lactate production and glucose consumption in HepG2 cells with or without esculetin. It was found that GPD2 bound strongly to GPI, revealing the direct interaction between the two glycolysis-related proteins. Animal tests have further demonstrated that esculetin may have anticancer effects by affecting the activity of PGK2, GPD2, and GPI. The results of this study demonstrated that esculetin can affect the glucose metabolism by binding to glycolytic proteins, thus playing an anti-tumor role, and these proteins which have direct interactions are potential novel targets for tumor treatment by esculetin.

The high expression of fatty acid synthase (FAS) in tumor cells is consistent with their elevated requirement for fatty acids for cell membrane synthesis and energy supply to support their almost unlimited proliferation. The expression levels of FAS in tumor cells are related to their proliferation, invasion, and metastasis. This study investigated the possible bioactive ingredients (fraxin, esculetin, scopolin et al.) of Cortex Fraxini and their effects on the interaction between specific proteins. We used microscale thermophoresis (MST) to show that our target protein, FAS (screened by combining transcriptome and network pharmacology), bound to the active compounds in Cortex Fraxini. It was found that FAS bound strongly to Glucose-6-phosphate isomerase (GPI), and that scopolin could affect this interaction by proteomics and MST. The results of this study demonstrate that the active compounds in Cortex Fraxini could play an anti-tumor role by binding to FAS and inhibiting the interactions between FAS and GPI to affect glucose and lipid metabolism, and that the protein pathway is a potential novel target for tumor treatment.

This study explored the possible bioactive ingredients and target protein of Rostellularia procumbens (L.) Nees. The results of optical turbidimetry revealed that the ethyl acetate extraction obtained from R. procumbens (L.) Nees could inhibit platelet aggregation. Gene chip was used to investigate differentially expressed genes. According to the results of the gene chip, the targets of compounds isolated from the ethyl acetate extraction were predicted by network pharmacology. Computational studies revealed that chinensinaphthol methyl ether and neojusticin B may target the integrin α_IIbβ₃ protein. The results of Prometheus NT.48 and microscale thermophoresis suggested that the molecular interactions between the two compounds with purified integrin α_IIbβ₃ protein in the optimal test conditions were coherent with the docking results. To our best knowledge, this is the first report to state that chinensinaphthol methyl ether and neojusticin B target the integrin α_IIbβ₃ protein.

The present study explored the possible bioactive ingredients and target protein of Rostellularia procumbens (L.) Nees. Firstly, we found that the ethyl acetate extraction obtained from R. procumbens could inhibit platelet aggregation. Then, gene chip was used to investigate differentially expressed genes and blood absorption compounds were investigated using high performance liquid chromatography-mass spectrometry characterization (LC-MS). Depending on the results of gene chip and LC-MS, the targets of blood absorption compounds were predicted according to the reverse pharmacophore matching model. The platelet aggregation-related genes were discovered in databases, and antiplatelet aggregation-related gene targets were selected through comparison. The functions of target genes and related pathways were analyzed and screened using the DAVID database, and the network was constructed using Cytoscape software. We found that integrin α_IIbβ₃ had a highest degree, and it was almost the intersection of all pathways. Then, blood absorption compounds were screened by optical turbidimetry. Western blot (WB) revealed that justicidin B separated from the ethyl acetate fraction may inhibit the expression of integrin α_IIbβ₃ protein. For the first time, we used Prometheus NT.48 and MST to detect the stability of this membrane protein to optimize the buffer and studied the interaction of justicidin B with its target protein. To our best knowledge, this is the first report to state that justicidin B targets the integrin α_IIbβ₃ protein. We believe that our findings can provide a novel target protein for the further understanding of the mechanism of R. procumbens on platelet aggregation.

Background/Objectives: Isoliquiritigenin (ISL) possesses excellent anti–inflammatory and colonic mucosal repair activities, showing promising potential for colitis treatment. However, the clinical application of native ISL is severely restricted by its poor water solubility, low in vivo bioavailability, and insufficient colon tissue accumulation. To solve these inherent delivery defects of free ISL, this study aimed to construct a novel oral colon–targeted solid nanoparticle–hydrogel composite delivery system and explore its therapeutic efficacy and underlying mechanisms for colitis therapy. Methods: ISL–loaded poly (lactic-co–glycolic acid)/polycaprolactone solid nanoparticles (IPP NPs) were first fabricated to improve the water solubility and in vivo delivery efficiency of free ISL. Thereafter, IPP NPs were encapsulated into a composite hydrogel network consisting of sodium alginate, chitosan, and squid ink polysaccharide to prepare the colon–targeted IPP@Gels. The pH–responsive drug release behavior of the composite system in the upper gastrointestinal tract was verified via in vitro characterization experiments. A dextran sulfate sodium (DSS)–induced murine colitis model was established to evaluate the in vivo therapeutic effects of IPP@Gels. The regulatory effects of the delivery system on intestinal tight junction protein expression, proinflammatory cytokine levels, gut microbiota composition, and short–chain fatty acid (SCFA) content were further investigated. Results: The constructed IPP@Gels exhibited favorable pH–responsive performance, which effectively inhibited the premature release of ISL in the upper gastrointestinal tract and achieved precise colon localization and sustained drug release. In vivo experimental results demonstrated that IPP@Gels significantly alleviated the pathological symptoms of colitis in model mice. Mechanistically, the composite system prominently upregulated the expression of intestinal tight junction proteins, reduced the secretion of proinflammatory cytokines, effectively enriched ISL accumulation in colon tissues, reversed gut microbiota dysbiosis, and increased the content of intestinal SCFAs, thereby producing synergistic anti-colitis therapeutic effects. Conclusions: The constructed SA/CS/SIP composite hydrogel (IPP@Gels), embedded with ISL–loaded nanoparticles, exhibits favorable biocompatibility and pH-responsive release behavior, enabling colon-localized retention of ISL. This novel oral drug delivery system mitigates several delivery–related drawbacks of free ISL and confers beneficial therapeutic effects in experimental colitis. It provides a feasible oral formulation strategy for colon–localized delivery of poorly soluble drugs and offers promising prospects for the management of colitis.