Nano Acacetin Mitigates Intestinal Mucosal Injury in Sepsis Rats by Protecting Mitochondrial Function and Regulating TRX1 to Inhibit the NLRP3 Pyroptosis Pathway

Background: Acacetin (AC) is a flavonoid compound with antiperoxidant, anti-inflammatory, and antiplasmodial activity. However, the solubility of AC is poor and nano acacetin (Nano AC) was synthesized. The intestinal mucosal barrier is impaired in sepsis rats, and the protective effects and mechanism of AC and Nano AC on the intestinal mucosal barrier are unclear.

Methods: Cecal ligation and perforation (CLP) was used to induce sepsis in rats, and lipopolysaccharide (LPS)-stimulated intestinal epithelial cells were used to observe the effects of AC and our synthesized Nano AC on the amelioration of intestinal mucosal damage. The molecular docking technique was used to predict the binding energy of AC to thioredoxin reductase 1 (TRX1) signaling pathway proteins. TRX1 inhibitor (PX-12) was employed to elucidate the protective signaling pathway of Nano AC in LPS-stimulated intestinal epithelial cells.

Results: Our synthesized Nano AC, with an average particle size of 17.18 ± 0.48 nm and an uptake rate of 95% in intestinal epithelial cells. The maximum binding capacity of AC to TRX1 was -6.82 kcal/mol, supporting the hypothesis that TRX1 is a potential target of AC. AC and Nano AC ameliorated the survival rate, intestinal mucosal damage score, pathological morphology, hepatic and renal function, and myocardial troponin levels, decreased serum levels of pyroptosis-related factors, upregulated TRX1, down-regulated NOD-like receptor protein 3 inflammasome (NLRP3), cysteinyl aspartate specific proteinase-11 (Caspase-11), Gasdermin D (GSDMD) in sepsis rats. They improved mitochondrial morphology and mitochondrial reactive oxygen species (ROS) levels, reduced pyroptosis levels, and upregulated TRX1, which adjusted NLRP3/ Caspase-11/ GSDMD signaling pathway in LPS-stimulated intestinal epithelial cells. Moreover, Nano AC was more effective.

Conclusion: AC and Nano-AC can inhibit the NLRP3/Caspase-11/GSDMD signaling pathway by upregulating TRX1 to ameliorate intestinal mucosal injury in sepsis rats, and the effect of Nano AC is more prominent.