Background
Ferroptosis, a novel manner of cell death depended on iron ion, contributed to goat mammary epithelial cell dysfunction. Interleukin-6 (IL-6) is a major pro-inflammatory factor during many inflammation-related diseases including mastitis, and a quite recently identified ferroptosis inducer. This study aims to explore the role of IL-6 in the dysfunction of goat mammary epithelial cells (GMECs) and how the level of IL-6 was regulated.
Methods
Primary GMECs were isolated, cultured and treated with lipopolysaccharide (LPS) alone or together with Ferrostatin-1 (Fer-1), a well-known ferroptosis inhibitor. CCK-8 was used to detect cell viability, ELISA was used to detect TNF-α content, and the levels of ROS, GSH and MDA were analyzed with DCFDA-cell ROS detection kit, GSH assay kit and MDA assay kit, respectively. The iron ion level was measured with an iron assay kit.
Results
The expression level of IL-6 protein in GMECs was up-regulated in response to LPS treatment, and the secretion of TNF-α, the cell oxidative stress level and the Fe2+ ion content was robustly increased, which could be reversed by Fer-1 treatment. Knockdown of IL-6 decreased cell oxidative stress level and inhibited ferroptosis in LPS-treated GMECs. Further, ubiquitin experiment and co-immunoprecipitation assay showed that USP14 upregulated IL-6 protein expression by reducing the ubiquitination of IL-6, and overexpression of IL-6 reversed the inhibitory effect of USP14 shRNA on LPS-treated GMECs ferroptosis. The NRF2 inhibitor Brusatol reversed the inhibitory effect of IL-6 shRNA on LPS-treated ferroptosis.
Conclusion
IL-6 protein is deubiquitinated by USP14 and upregulated in LPS-treated GMECs, further promoting ferroptosis and inflammation through the NRF2 signaling pathway.
Circularly polarized luminescence (CPL)-active materials have attracted increasing attention due to their applications in chiral sensing and optoelectronics. However, it is still a challenging task to fabricate CPL materials with large luminescence dissymmetry factors and strong emission intensities in the solid state. In this work, two singlehanded twisted tetraarylethylene (TAE)-bridged polybissilsesquioxane nanotubes have been facilely constructed through a supramolecular templating polymerization of TAE-bridged bis(triethoxysilane) with aggregation-induced emission properties, using self-assemblies of a pair of chiral amino acid-based cationic amphiphiles as the supramolecular templates. It is found that the chirality has been transferred from the self-assemblies to the polybissilsesquioxane nanotubes. The resulting hybrid silicas show high thermal stabilities and bright CPL with the fluorescence quantum yield up to 62.8%.
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