Department of Hepatobiliary Surgery, The First Affiliated Hospital of Chongqing Medical University, Chongqing 400016, China.
Department of Anatomy, Basic Medical College, Chongqing Medical University, Chongqing 400016, China.
Hepatobiliary Pancreat Dis Int. 2021 Aug;20(4):352-360. doi: 10.1016/j.hbpd.2021.04.013. Epub 2021 May 8.
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.
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.
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.
Fisetin showed protective effects against hepatic IRI, countering inflammatory responses through mediating the GSK3β/AMPK/NLRP3 inflammasome pathway.
肝缺血再灌注（IRI）损伤是肝移植的一个关键挑战。漆黄素具有抗炎、抗衰老和抗氧化作用。本研究旨在探讨漆黄素是否能减轻肝 IRI，并探讨其潜在机制。
对假手术或温热性肝 I/R 手术的小鼠用漆黄素（5、10 或 20mg/kg）预处理。进行肝组织学评估、TUNEL 检测和血清转氨酶测量。还使用 RAW264.7 巨噬细胞进行体外低氧/复氧（H/R）模型，并用漆黄素（2.5、5 或 10µmol/L）预处理。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 Western blot 检测 p-GSK3β、p-AMPK 和 NOD、LRP 和热蛋白结构域包含 3（NLRP3）相关蛋白的水平。
与 I/R 组相比，漆黄素预处理减轻了小鼠 IRI 模型的肝损伤、血清转氨酶水平、血清中 IL-1β、IL-18 和 TNF-α的浓度。漆黄素还降低了 I/R 手术肝中 NLRP3 炎性体相关蛋白（NLRP3、裂解半胱氨酸蛋白酶-1、IL-1β和 IL-18）的表达。体外实验表明，漆黄素降低了 H/R 处理的 RAW264.7 细胞中 IL-1β、IL-18 和 TNF-α的释放，并降低了 NLRP3 炎性体相关蛋白的表达。此外，漆黄素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 信号通路。AMPK 特异性抑制剂化合物 C 部分抑制了漆黄素的抗炎作用。
漆黄素对肝 IRI 具有保护作用，通过调节 GSK3β/AMPK/NLRP3 炎性体通路对抗炎症反应。
