Bcl-2 Recombinant Mouse Monoclonal Antibody [M43D02]

BackgroundDry eye is a multifactorial ocular surface disorder in which tear hyperosmolarity acts as a major stressor that promotes inflammatory injury and apoptosis in corneal epithelial cells. Wogonoside (WGS), a flavonoid glycoside derived from Scutellaria baicalensis, has reported anti-inflammatory and cytoprotective properties; however, its role and mechanism in dry eye–related epithelial injury remain insufficiently defined.MethodsNetwork pharmacology was used to identify putative targets shared by WGS and dry eye, followed by pathway enrichment analysis. Molecular docking, molecular dynamics simulation, and cellular thermal shift assay (CETSA) were further used to evaluate the interaction between WGS and AKT1. Experimental validation was performed in transformed human corneal epithelial cells (HCE-T cells) exposed to hyperosmotic medium (500 mOsm). Cell viability, proliferation, apoptosis, inflammatory mediator levels, and PI3K/AKT signaling-related changes were assessed. To interrogate pathway involvement, cells were additionally treated with the AKT activator SC79 (10 μM) or the AKT inhibitor MK-2206 (2.5 μM).ResultsA total of 203 overlapping targets were identified between WGS-related and dry eye-related target sets. Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment highlighted PI3K/AKT signaling as a potentially relevant pathway, and AKT1 was prioritized as a candidate target. Molecular docking and molecular dynamics simulation supported a stable interaction between WGS and AKT1, while CETSA provided additional evidence of AKT1 target engagement in cells. In hyperosmotic HCE-T cells, WGS improved cell viability and proliferative capacity, attenuated apoptosis, and reduced interleukin-1 beta (IL-1β), interleukin-6 (IL-6), and matrix metalloproteinase-9 (MMP-9) levels. These effects were accompanied by restoration of PI3K/AKT signaling. Mechanistically, MK-2206 partially attenuated the protective effects of WGS, whereas SC79 produced a similar protective profile, supporting a PI3K/AKT-dependent component.ConclusionWogonoside alleviates hyperosmotic stress-induced inflammatory injury and apoptosis in HCE-T cells. Integrated evidence from network pharmacology, molecular docking, molecular dynamics simulation, CETSA, and in vitro validation supports AKT1 as a candidate target of WGS. The protective effects of WGS were closely associated with restoration of PI3K/AKT signaling, suggesting that WGS may be a promising candidate for hyperosmotic stress-related corneal epithelial injury and merits further validation in appropriate animal models of dry eye.

Oligoasthenospermia (OAS), a major cause of male infertility, was alleviated by Guilu-Erxian-Glue (GLEXG) in Tripterygium wilfordii polyglycoside (TWG)-induced rat models. This study identified GLEXG's bioactive ingredients and explored its therapeutic mechanism. Using network pharmacology and liquid chromatography-mass spectrometry, quercetin was predicted and validated as a key component of GLEXG. In vivo and in vitro, OAS models were established using TWG in mice and H 2 O 2 in spermatocytes. TWG administration reduced testicle and epididymis indices, sperm concentration, vitality, and serum testosterone, luteinizing hormone, and follicle-stimulating hormone. It also increased germ cell apoptosis, upregulated Bax, cleaved caspase-3, and cleaved caspase-9, downregualted Bcl-2, PHB1, VDHC, SDHA, and CoxIV, and impaired mitochondrial function (reduced mitochondrial DNA copy number and membrane potential). Quercetin counteracted these effects in vivo and in vitro. Mechanistically, quercetin increased p-PKA/PKA and p-CREB/CREB ratios, upregulated Sirt1, and decreased Ac-p53/p53 ratio. These beneficial effects were abolished upon Sirt1 inhibition. Thus, quercetin in GLEXG ameliorated OAS by attenuating apoptosis and mitochondrial dysfunction via PKA/CREB pathway-dependent activation of Sirt1 and deacetylation of p53.

Background Ferroptosis plays a key role in the development of chronic obstructive pulmonary disease (COPD). Whether ginsenoside Rg1 improves cigarette smoke-induced COPD or whether ginsenoside Rg1 improves COPD by inhibiting ferroptosis remains unknown. Methods BEAS-2B cells were exposed to cigarette solution (CSE) for 24 hours and treated with ginsenoside Rg1, the ferroptosis inhibitor Fer-1, and the PERK inhibitor GSK. Cell viability, endoplasmic reticulum stress, mitochondrial morphology, membrane potential, reactive oxygen species (ROS), iron levels, and the expression of related proteins were detected using corresponding methods. A COPD mouse model was constructed using cigarette smoke (CS). Ginsenoside Rg1 and GSK were administered via tube feeding 15 days after successful modeling. Mouse lung tissues were evaluated by HE staining. The expression of inflammatory markers, ROS, iron content, and related proteins was detected using corresponding methods. Results The results demonstrated that in the CSE-exposed BEAS-2B cell model and CS-induced mouse COPD model, the expression levels of endoplasmic reticulum stress (ERS)-related factors such as GRP78 were increased, while those of the antioxidant markers GPX4 and GSH were significantly decreased. Ginsenoside Rg1 improved emphysema and inflammation by inhibiting ferroptosis in vivo and in vitro. Using a PERK inhibitor, we found that ginsenoside Rg1 inhibited ferroptosis in vivo and in vitro by regulating ERS. Conclusion This study showed that ginsenoside Rg1 alleviates cigarette smoke-induced COPD by regulating the PERK/ATF4 axis to inhibit ERS and ferroptosis.