NLRP3 inhibitor(Kodiak Sciences)

Long-term excessive ethanol consumption is known to accelerate the progression of diabetes mellitus (DM). Our previous study indicated that salidroside (SAL) improves hepatic insulin resistance in DM mice. However, the underlying mechanisms of ethanol on this process and whether SAL can exert a therapeutic effect are not fully understood. The aim of this study is to investigate the effect of SAL on ethanol-exacerbated hepatic insulin resistance and its underlying mechanisms in DM conditions. Primary mouse hepatocytes were isolated and cultured to establish an insulin sensitivity impaired model. The effects of ethanol exposure at different concentrations or for different durations on intracellular lipid accumulation, the insulin signaling pathway of Akt/glycogen synthase kinase 3β (GSK3β), and the NACHT, LRR and PYD domains-containing protein 3 (NLRP3) inflammasome signaling were observed. An NLRP3 inhibitor was used to determine whether the effects of ethanol were dependent on the overactivation of the NLRP3 inflammasome signaling and to investigate the intervention effects of SAL. A DM model was established by feeding a high-fat diet combined with streptozotocin injection to observe the effects of ethanol exposure on blood glucose and body weight, biochemical and histopathological indicators, as well as the Akt/GSK3β and NLRP3 inflammasome signaling in liver tissue. In vitro, ethanol exposure exacerbated lipid accumulation and insulin sensitivity impairment in hepatocytes in a concentration- and time-dependent manner. This effect was dependent on the overactivation of the NLRP3 inflammasome, and co-incubation with SAL could effectively improve the above phenomena. In vivo, ethanol exposure also exacerbated liver insulin resistance, and SAL treatment could improve this condition. SAL has an ameliorative effect on the ethanol-exacerbated liver insulin resistance in diabetic mice.

Ischemic stroke, a severe neurological disorder with a multifactorial pathogenesis, presents significant therapeutic challenges. Calycosin, a natural flavonoid, has diverse biological activities, including antioxidant, anti-inflammatory, and antitumor effects. In this study we investigate the protective effects of calycosin against blood-brain barrier (BBB) damage following cerebral ischemia-reperfusion injury (CIRI) and explore the underlying mechanisms. We employ middle cerebral artery occlusion (MCAO) in rats and oxygen-glucose deprivation (OGD) in bEnd.3 brain microvascular endothelial cells to assess neurological function, BBB integrity, the expression of pyroptosis-related proteins, inflammatory mediator release, endothelial barrier permeability, and cell viability. The results reveal that calycosin significantly ameliorates CIRI-induced BBB damage, as evidenced by improved neurological scores, reduced brain water content, and decreased infarct volume. Calycosin suppresses NLRP3-mediated pyroptosis by downregulating HMGB1, NLRP3, caspase 1, GSDMD, N-GSDMD, and IL-18 expression while reducing the secretion of HMGB1, IL-1β, and IL-18. Additionally, calycosin enhances BBB integrity by decreasing MMP9 and AQP-4 expression and upregulating the expression of tight junction proteins (ZO-1, occludin, and claudin-5). In OGD-treated bEnd.3 cells, calycosin inhibits NLRP3-mediated pyroptosis, reduces inflammatory mediator release, and improves cell viability and barrier function. Notably, molecular docking and molecular dynamics simulations demonstrate that calycosin stably binds to NLRP3 with high affinity, supporting its potential as an NLRP3 inhibitor. These findings indicate that calycosin protects against CIRI-induced BBB damage by inhibiting NLRP3-mediated pyroptosis and modulating tight junction protein expression, indicating that calycosin is a potential therapeutic option for ischemic stroke.