Clinical and Experimental Pharmacology and Physiology最新文献

Natural products and their semisynthetic derivatives possess tremendous medicinal properties and have the potential to modulate the immune system, providing new therapeutic options for drug development. In this study, we evaluated Dehydrozingerone-15, a novel dehydrozingerone derivative, for its anti-inflammatory and antioxidant properties through standardised in vitro and in vivo approaches. Dehydrozingerone-15 suppressed the stimulatory effect of LPS in RAW 264.7 cells by reducing the secretion of interleukin-6 (IL-6), tumour necrosis factor-alpha (TNF-α), interleukin-1 beta (IL-1β), interleukin-2 (IL-2) and nitric oxide. Western blot analysis at the mechanistic level showed a reduced expression level of nitric oxide synthase (iNOS), IκB kinase beta (Ikk-β) and nuclear factor kappa-B (NF-κB/p65). Confocal microscopy studies further demonstrated that Dehydrozingerone-15 reduced the expression of NF-κB/p65 markedly. In the in vivo LPS-induced sepsis model, Dehydrozingerone-15 administration reduced TNF-α and IL-6 expression and protected vital organs (lungs, kidneys, and liver) from acute inflammation. The anti-inflammatory potential of Dehydrozingerone-15 was further validated in leukocyte migration induced by carrageenan and vascular permeability triggered by acetic acid assays, both of which showed significant inhibition. Pharmacokinetic analysis revealed that Dehydrozingerone-15 was rapidly absorbed in BALB/c mice, reaching a C_max of 10 349 ng/mL at 0.25 h. The total drug exposure (AUC_0–α) was 13 862 ng.h/mL, indicating sustained exposure, with high tissue distribution (20 L/kg) and moderate clearance. Additionally, toxicological evaluation at doses up to 2000 mg/kg body weight showed no significant alterations in haematological parameters compared with the vehicle control. Furthermore, based on a comparative evaluation of in vitro and in vivo results, Dehydrozingerone-15, relative to its parent molecule, demonstrates significant therapeutic potential with high efficacy against inflammation and oxidative stress.

Postoperative peritoneal adhesion (PA) formation is a common complication after abdominal surgery and can result in various severe outcomes. Inflammation and fibrosis are important processes in PA formation. The effectiveness of kaempferol (KF), a common component of several medications used to reduce inflammation and prevent fibrotic diseases, in preventing postoperative PA formation is unknown. This study explored the effectiveness and mechanism of KF in preventing PA formation following surgery. The animal adhesion model revealed that KF could effectively prevent adhesions formation and inhibit mesothelial–mesenchymal transition (MMT). Protein–protein interaction and pathway enrichment analyses revealed that TNF-α may be the key target through which KF prevents adhesion formation. Here, KF was found to inhibit TNF-α-induced MMT. Furthermore, Kyoto Encyclopedia of Genes and Genomes pathway enrichment analyses revealed that common genes between KF and PA are enriched in the TNF signalling pathway. Moreover, cyclooxygenase 2 (COX2) was identified as a downstream target of TNF-α whose expression is positively correlated with adhesion formation. Most importantly, COX2 small interfering RNA (siRNA) and overexpression plasmid (OE) transfection experiments confirmed that KF inhibits MMT by blocking the TNF-α/COX2 signalling pathway. Finally, molecular docking revealed that TNF-α is a binding target of KF. In conclusion, these results suggest that KF inhibits MMT by blocking the TNF-α/COX2 signalling pathway, thus attenuating adhesion formation. These results provide new insights into the development of antiadhesion drugs.

Molecular glues have emerged as a novel class of targeted protein degraders with broad potential in cancer therapy. By inducing proximity between E3 ubiquitin ligases and oncogenic proteins, these agents activate the ubiquitin–proteasome system to drive selective protein degradation. This review systematically explores the application strategies of molecular glues in the targeted degradation of cancer-related proteins located in different subcellular compartments: nuclear cancer proteins (Ikaros, Helios, Aiolos, B-cell lymphoma 6 protein, cyclin K, RNA binding motif protein 39) and cytoplasmic cancer proteins (β-catenin, casein kinase 1α, G1 to S phase transition factor 1). Additionally, this work discusses current challenges and optimisation strategies, offering new perspectives for the development of precision anti-cancer therapeutics.

This study aims to investigate the therapeutic efficacy and molecular mechanism of cinobufagin in non-small cell lung cancer (NSCLC) via pyroptosis induction. Bronchial epithelial cells and NSCLC cell lines were treated with gradient concentrations of cinobufagin. Cell viability was evaluated using Cell Counting Kit-8 (CCK-8) assay. RNA-sequencing was performed to identify differentially expressed genes. Lactate dehydrogenase (LDH) release was measured via cytotoxicity detection kit. Pyroptotic morphological changes were observed by transmission electron microscopy. Western blotting analysed expression levels of pyroptosis-related proteins. In vivo efficacy was validated in nude mouse xenograft models. Immunohistochemistry evaluated tumour pyroptosis markers, whilst flow cytometry analysed tumour-infiltrating CD8⁺ T cells and natural killer (NK) cells. Cinobufagin demonstrated selective cytotoxicity against NSCLC cells with minimal toxicity to normal bronchial epithelium. RNA-seq analysis revealed significant enrichment of pyroptosis-related pathways. Functional experiments confirmed cinobufagin-induced LDH release, characteristic pyroptotic morphological changes and upregulation of cleaved caspase-3 and Gasdermin E (GSDME)-NT in NSCLC cells. In xenograft models, cinobufagin treatment reduced tumour volume compared to controls. Mechanistically, this was associated with enhanced caspase-3 activation and GSDME-NT accumulation in tumour tissues. Notably, cinobufagin treatment significantly increased NK cell infiltration and activity. Cinobufagin exerts antitumor effects in NSCLC through caspase-3/GSDME-mediated pyroptosis induction, accompanied by immune microenvironment modulation. These findings provide preclinical evidence for cinobufagin as a potential therapeutic agent targeting pyroptosis in NSCLC.

Sinomenine (SIN) is an extract obtained from the plant Sinomenium acutum, which has been reported in hepatocellular carcinoma (HCC) and many other different types of human cancers as an anti-tumour agent. However, the molecular mechanism modulated or altered by SIN in HCC progression remains to be investigated. This study aims to uncover the molecular mechanism contributing to the tumour-suppressing effect of SIN in HCC. At first, HCC cells were treated with SIN for functional detections. Results of functional assays demonstrated that SIN was an efficient inhibitor of HCC cell viability, migration, and invasion. Next, O-GlcNAcylation and its removal OGA were found to be regulated by SIN treatment in HCC cells. According to rescue functional assays, OGA knockdown strengthened HCC cell viability, migration, and invasion suppressed by SIN. Moreover, as detected by mechanism experiments, OGA directly interacted with SP1 and blocked SP1 O-GlcNAcylation at site T407, indicating that SP1 was downregulated by OGA-mediated deglycosylation. Additionally, silencing of SP1 recovered the suppressing effects of SIN on HCC cell proliferation, migration, and invasion attenuated by OGA knockdown. Finally, SIN treatment inhibited the tumour tissue growth in vivo. In summary, this study unmasked the anti-tumour effect of SIN in HCC and SIN exerted functions by regulating OGA-mediated downregulation of SP1.

Macrophage infiltration was closely associated with inflammatory injury of podocytes in diabetic nephropathy (DN), while how macrophages affected podocytes remained not entirely clear. Here, we not only investigated the relationship between macrophages and high glucose (HG)-treated podocytes, but the underlying mechanisms were also explored. Transmission electron microscopy, nanoparticle tracking analysis, and western blot of CD9, CD63, CD81, and Calnexin were performed to identify exosomes; QRT-PCR was performed to detect AFAP1-AS1 expression; Western blot was performed to examine NLRP3, Cleaved caspase-1, and GSDMD-N protein levels; Immunofluorescence was performed to assess co-localisation of NLRP3 and ASC; ELISA was performed to detect IL-18 and IL-1β levels; Cytotoxicity LDH Assay Kit was performed to detect LDH level; RNA pulldown was performed to determine the interaction of AFAP1-AS1 and EZH2; ChIP was employed to determine the interaction of EZH2 and H3K27me3 in the NLRP3 promoter region. The results showed that AFAP1-AS1 expression was down-regulated in the peripheral blood of DN patients, and exosomes derived from M2 macrophages transfected with si-AFAP1-AS1 enhanced HG-induced podocyte pyroptosis via significantly elevating NLRP3, Cleaved caspase-1, and GSDMD-N protein levels, immunofluorescence intensity of NLRP3 and ASC, as well as IL-18, IL-1β, and LDH levels. Mechanistically, AFAP1-AS1 interacted with EZH2 to transcriptionally regulate H3K27me3 level in the NLRP3 promoter region, thus epigenetically repressing NLRP3 level to inhibit podocyte pyroptosis. These results may provide an important target for improving kidney injury in DN.