Role and mechanism of ferroptosis mediated ischemia reperfusion injury in steatosis liver
10.3760/cma.j.cn421203-20221227-00340
- VernacularTitle:小鼠脂肪肝缺血再灌注损伤中铁死亡的作用及机制研究
- Author:
Yuhan XIA
1
;
Jingyang GU
;
Jian DENG
;
Jiandong WANG
Author Information
1. 上海交通大学医学院附属新华医院普外科，上海 200092
- Keywords:
steatosis liver;
ischemia-reperfusion injury;
ferroptosis
- From:
Chinese Journal of Organ Transplantation
2023;44(8):487-495
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To explore the phenotype and mechanism of ferroptosis in steatosis liver ischemia reperfusion injury (IRI) and elucidate the mechanism of targeted inhibition of ferroptosis down-regulating steatosis liver IRI.Methods:First, 20 mice are divided into 2 group according to the different feeding mode: Normal chow diet(NCD group, n=10)and high fat diet(HFD group, n=10). The fatty liver constructs are successful as verified by laboratory tests for relevant indicators and oil red staining.NCD and HFD mice were randomized into two groups sham and IRI.The expression of glutathione peroxidase 4(Gpx4)in liver tissues of each group is detected by Western blot.Morphological phenotypes of mitochondria in liver tissue are observed by transmission electron microscope.Accordingly, it was determined whether ferroptosis occurred in mouse liver IRIs.NCD(n=30)and HFD(n=30)are further randomized into 3 groups: sham(n=10), IRI(n=10)and inhibitor(IRI-fer-1 group, n=10). The serum contents of alanine aminotransferase(ALT)and aspartate aminotransferase(AST)are detected by ALT/AST kits.Hematoxylin-eosin(HE)stain is employed for detecting the degree of liver injury; immunohistochemistry(IHC)and enzyme-linked immunosorbent assay(ELISA)for detecting the levels of inflammatory factors and inflammatory cell infiltration.Similarly, primary hepatocytes from NCD/HFD mice are extracted and the occurrence of ferroptosis is verified by detecting C11-BODIPY(581/591)by fluorescent stain after hypoxia-reoxygenation(H/R).Results:As compared with NCD-IRI group, Gpx4 protein expression declined obviously in HFD-IRI group while serum ALT/AST level spiked markedly( P<0.01). IHC staining of 4-HNE is positive, mitochondrial specific damage is more pronounced and inflammatory infiltration became enhanced.As compared with IRI group, serum level of ALT/AST dropped obviously and infiltration of inflammatory cells and secretion of inflammatory factors are blunted markedly in IRI-fer-1 group( P<0.01). Conclusions:A high degree of ferroptosis and severe inflammatory response during fatty liver IR are features of distinguishing steatosis liver IRI from ordinary liver IRI.Targeted inhibition of ferroptosis lowers inflammation and damage during IR in steatosis liver.
