Estrogen alleviates hepatocyte necroptosis depending on GPER in hepatic ischemia reperfusion injury

J Physiol Biochem. 2022 Feb;78(1):125-137. doi: 10.1007/s13105-021-00846-5. Epub 2021 Oct 15.

Abstract

Hepatic ischemia reperfusion injury (IRI) occurs in liver transplantation, complex liver resection, and hemorrhagic shock, which causes donor organ shortage and hepatic damage. The burst of reactive oxygen species (ROS) during reperfusion leads to cell apoptosis and necroptosis. It has been reported that estrogen could attenuate hepatic IRI. G protein estrogen receptor (GPER) mediates estrogen effects via nonclassic receptor systems. Here, we investigate whether estrogen protecting liver from hepatic IRI depends on GPER and the influence of GPER activation on hepatocyte necroptosis. We proved that estrogen had a protective effect on both hepatocyte hypoxia re-oxygen (H/R) challenge and mouse hepatic ischemia reperfusion model. However, the application of GPER specific antagonist G15 before estrogen inhibited this beneficial effect. The results of mitochondria functional measurement revealed that estrogen improved hepatocyte mitochondria function by activating GPER, which might benefit from the increased expression of connexin 43 (Cx43) in mitochondria. To investigate the relationship between GPER activation and necroptosis, we used caspase-3/7 inhibitor benzyloxycarbonyl-Asp(OMe)-Glu(OMe)-Val-Asp(OMe)-chloromethylketone (Z-DEVD-FMK) to eliminate the interference of apoptosis. Estrogen showed a protective effect on hepatic IRI after using Z-DEVD-FMK, which could be suppressed by G15. GPER activation decreased the level of receptor interacting protein kinase (RIPK) 3, phosphorylated (p-) RIPK1, and p-mixed lineage kinase domain-like (MLKL). The co-immunoprecipitation result indicated that GPER could bind with RIPK3. GPER is indispensable in estrogen protecting liver from IRI. GPER activation attenuates hepatocyte necroptosis by decreasing the level of RIPK3, p-RIPK1, and p-MLKL.

Keywords: Estrogen; GPER; Hepatic ischemia reperfusion injury; Necroptosis.

MeSH terms

  • Animals
  • Estrogens
  • GTP-Binding Proteins / metabolism
  • Hepatocytes / metabolism
  • Liver / metabolism
  • Mice
  • Necroptosis*
  • Receptors, Estrogen / metabolism
  • Reperfusion Injury* / metabolism
  • Reperfusion Injury* / prevention & control

Substances

  • Estrogens
  • Receptors, Estrogen
  • GTP-Binding Proteins