Clinical and Experimental Pharmacology and Physiology最新文献

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.

The photoperiod is a crucial factor affecting the seasonal rhythms of mammals. Under the seasonal rhythms, the growth and development of the body are closely related to the oxidative respiratory function of mitochondria, but the influence of the single seasonal factor photoperiod on it remains unclear. In this study, mitochondrial dynamics and associated regulatory mechanisms were investigated in the livers of striped dwarf hamsters (Cricetulus barabensis) exposed to three distinct photoperiod regimes: short photoperiod (SP), moderate photoperiod (MP) and long photoperiod (LP). Results indicated that: (1) Liver mass responses to photoperiod varied with changes in body weight. (2) ATP synthase activity was significantly decreased under LP, whereas citrate synthase activity (CS) was selectively diminished under SP. (3) The Bcl2 associated X protein (bax) to b cell lymphoma 2 (bcl2) ratio increased under both SP and LP. (4) Dynamin-related protein 1 (DRP1) protein abundance increased under both LP and SP conditions, while mitochondrial fission factor (MFF) decreased under LP, signifying divergent remodelling of mitochondrial fission signalling. (5) Caspase3 activity unchanged under SP. In conclusion, under LP and SP treatment, the oxidative respiratory function of liver mitochondria in striped dwarf hamsters may be weakened, potentially due to increased mitochondrial membrane permeability, as indicated by an elevated bax/bcl2 ratio. Compared to long photoperiod treatment, the upregulation of the liver mitochondrial fission signalling and the lower level of apoptosis under short photoperiod conditions may help facilitate thermogenic adaptation to increased energetic demands in winter.

Abdominal aortic aneurysm (AAA), a vascular condition that endangers life, is typified by the progressive weakening and dilation of the aortic wall. In its pathogenic process, oxidative stress and angiogenesis assume crucial functions. This research explored the function of G protein-coupled receptor 39 (GPR39) in angiotensin II (AngII)-induced AAA in ApoE−/− mice and its underlying mechanisms. A series of in vivo and in vitro experiments were performed, including AngII infusion-induced AAA in mice, histological analysis, ELISA, Western blotting, quantitative PCR, and angiogenic tube formation assays. GPR39 expression was significantly downregulated in arterial tissues of AngII-induced AAA mice, as evidenced by reduced mRNA and protein. Administration of the GPR39 agonist TC-G 1008 markedly attenuated AAA progression, reducing aneurysm incidence and size. Furthermore, TC-G 1008 alleviated oxidative stress through reducing the levels of malondialdehyde (MDA) and augmenting the activity of superoxide dismutase (SOD) as well as the levels of glutathione (GSH) in arterial tissues. TC-G 1008 also suppressed the vascular endothelial growth factor A (VEGF-A) and vascular endothelial growth factor receptor 2 (VEGF-R2) expressions in both saline- and AngII-infused mice. In vitro, TC-G 1008 inhibited AngII-induced angiogenic tube formation in human umbilical vein endothelial cells (HUVECs) and downregulated VEGF-A/VEGFR2 signalling. Mechanistically, TC-G 1008 reduced the expression of the transcription factor Ets-related transcription factor 1 (ETS-1), which is involved in VEGF-A/VEGFR2 regulation. Overexpression of ETS-1 reversed the inhibitory effects of TC-G 1008 on VEGF-A/VEGFR2 levels and angiogenic tube formation, confirming the critical role of ETS-1 in this pathway. These findings demonstrate that GPR39 activation by TC-G 1008 protects against AngII-induced AAA by mitigating oxidative stress, suppressing VEGF-A/VEGFR2 signalling, and inhibiting ETS-1-mediated angiogenesis.

Few studies have investigated blood glucose levels and complication management in elderly patients with type 2 diabetes (T2D) at community hospitals in China. The objective of this study was to investigate the factors influencing blood glucose control in elderly patients with T2D. One thousand one hundred and fifty elderly patients (age ≥ 65 years) with T2D were involved in the study to assess blood glucose control, health status (including comorbidities and cognitive status), complication management, and adherence to medication according to the guidelines of the American Diabetes Association. The FRAIL scale was used to screen for frailty syndrome in the elderly patients. Univariate and multivariate logistic regression analyses were used to investigate the factors affecting glucose control. Among the 1150 participants, 351 (30.52%) had poor glucose control. Frailty (odds ratio [OR]:2.546; 95% confidence interval [CI]: 1.267–5.117; p = 0.009), male sex (OR:0.679; 95% CI: 0.522–0.884; p = 0.004), and insulin treatment (OR: 0.229; 95% CI: 0.165–0.317; p < 0.001) were significantly independently associated with poor blood glucose control. In conclusion, for elderly patients with T2D, more attention should be paid to men, insulin therapy initiation and screening for frailty.

Alcoholism and alcohol misuse are major public health concerns, and chronic alcohol consumption negatively impacts memory and can lead to neurodegeneration and neuroinflammation. Neuroinflammation is a prevalent characteristic of all neurological diseases, induced by oxidative stress and resulting in impaired neuronal function. Thus, targeting neuroinflammation and oxidative stress can be a pivotal mechanism for attenuating memory impairment. In the current study, new isoxazolone derivatives were synthesized, and their synthetic properties were characterized using proton nuclear magnetic resonance (¹H NMR) and Fourier transform infrared (FTIR) spectroscopy. The molecular simulation studies were performed on triggering receptors expressed on myeloid cells-1 (TREM-1) and nucleotide-binding domain, leucine-rich-containing family, pyrin domain-containing-3 (NLRP3). Further, these derivatives were evaluated for neuroprotective potential in an ethanol-induced neuroinflammation model in male albino mice. Our results demonstrated ethanol-induced cognitive deficits, elevated inflammatory markers, ROS and altered endogenous antioxidant enzyme levels. At the same time, treatment with isoxazolone reversed the hyperactivated inflammatory markers, including NLRP3, and modulated the TREM-2 expression. Further, the antioxidant and anti-cholinesterase potential of these derivatives was also evaluated using in vitro assays. Our results suggested that these derivatives may help reduce neuroinflammation by acting on several stages of the inflammatory process in an ethanol-induced neurodegenerative model.