Background

Hepatic ischemia-reperfusion (I/R) injury (IRI) represents a crucial challenge in liver transplantation. Fisetin has anti-inflammatory, anti-aging and anti-oxidative properties. This study aimed to examine whether fisetin mitigates hepatic IRI and examine its underlying mechanisms.

Methods

Sham or warm hepatic I/R operated mice were pretreated with fisetin (5, 10 or 20 mg/kg). Hepatic histological assessments, TUNEL assays and serum aminotransferase measurements were performed. An in vitro hypoxia/reoxygenation (H/R) model using RAW264.7 macrophages pretreated with fisetin (2.5, 5 or 10 µmol/L) was also used. Serum and cell supernatant concentrations of interleukin-1β (IL-1β), IL-18 and tumor necrosis factor-α (TNF-α) were determined by enzyme-linked immunosorbent assay (ELISA). Protein levels of p-GSK3β, p-AMPK and NLR family pyrin domain-containing 3 (NLRP3)-associated proteins were detected by Western blotting.

Results

Compared with the I/R group, fisetin pretreatment reduced pathological liver damage, serum aminotransferase levels, serum concentrations of IL-1β, IL-18 and TNF-α in the murine IRI model. Fisetin also reduced the expression of NLRP3 inflammasome-associated proteins (NLRP3, cleaved caspase-1, IL-1β and IL-18) in I/R-operated liver. The experiments in vitro showed that fisetin decreased the release of IL-1β, IL-18 and TNF-α, and reduced the expression of NLRP3 inflammasome-associated proteins in H/R-treated RAW264.7 cells. Moreover, fisetin increased the expressions of p-GSK3β and p-AMPK in both models, indicating that its anti-inflammatory effects were dependent on GSK3β/AMPK signaling. The anti-inflammatory effects of fisetin were partially inhibited by the AMPK specific inhibitor compound C.

Conclusions

Fisetin showed protective effects against hepatic IRI, countering inflammatory responses through mediating the GSK3β/AMPK/NLRP3 inflammasome pathway.

中文翻译：

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非瑟酮通过调节 GSK3β/AMPK/NLRP3 炎性体通路减轻肝脏缺血再灌注损伤

背景

肝缺血再灌注 (I/R) 损伤 (IRI) 是肝移植中的一个关键挑战。非瑟酮具有抗炎、抗衰老和抗氧化特性。本研究旨在检查非瑟酮是否能减轻肝脏 IRI 并检查其潜在机制。

方法

用非瑟酮（5、10 或 20 mg/kg）预处理假手术或暖肝 I/R 手术小鼠。进行了肝组织学评估、TUNEL测定和血清转氨酶测量。还使用了使用用非瑟酮 (2.5、5 或 10 µmol/L) 预处理的 RAW264.7 巨噬细胞的体外缺氧/复氧 (H/R)模型。通过酶联免疫吸附试验（ELISA）测定血清和细胞上清液中白细胞介素-1β（IL-1β）、IL-18和肿瘤坏死因子-α（TNF-α）的浓度。通过蛋白质印迹检测 p-GSK3β、p-AMPK 和 NLR 家族 pyrin 结构域 3 (NLRP3) 相关蛋白的蛋白质水平。

结果

与 I/R 组相比，fisetin 预处理降低了小鼠 IRI 模型中的病理性肝损伤、血清转氨酶水平、血清 IL-1β、IL-18 和 TNF-α 浓度。Fisetin 还降低了 I/R 手术肝脏中 NLRP3 炎症小体相关蛋白（NLRP3、cleaved caspase-1、IL-1β 和 IL-18）的表达。体外实验表明，非瑟酮可降低 H/R 处理的 RAW264.7 细胞中 IL-1β、IL-18 和 TNF-α 的释放，并降低 NLRP3 炎性体相关蛋白的表达。此外，非瑟酮在两种模型中均增加了 p-GSK3β 和 p-AMPK 的表达，表明其抗炎作用依赖于 GSK3β/AMPK 信号传导。非瑟酮的抗炎作用被 AMPK 特异性抑制剂化合物 C 部分抑制。

结论

Fisetin 显示出对肝脏 IRI 的保护作用，通过介导 GSK3β/AMPK/NLRP3 炎性体通路对抗炎症反应。