G protein-coupled receptor 30-dependent protein kinase A pathway is critical in nongenomic effects of estrogen in attenuating liver injury after trauma-hemorrhage

Am J Pathol. 2007 Apr;170(4):1210-8. doi: 10.2353/ajpath.2007.060883.

Abstract

Although nongenomic effects of 17beta-estradiol (E2) are mediated via the estrogen receptor alpha (ER-alpha), the existence of another novel ER, G protein-coupled receptor 30 (GPR30), has been suggested as a candidate for triggering a broad range of E2-mediated signaling. GPR30 also acts independently of the ER to promote activation of the protein kinase A (PKA) pathway, which protects cells from apoptosis through Bcl-2. In this study, we examined whether the salutary effects of E2 in attenuating hepatic injury after trauma-hemorrhage are mediated via GPR30- or ER-alpha-regulated activation of PKA-dependent signaling. At 2 hours after trauma-hemorrhage, administration of E2-conjugated to bovine serum albumin (E2-BSA, membrane impermeable) or E2 induced the up-regulation of ER-alpha and GPR30 and attenuated hepatic injury. This was accompanied by increases in PKA activity and Bcl-2 expression. Inhibition of PKA in E2-BSA-treated trauma-hemorrhage rats by PKA inhibitor H89 prevented the E2-BSA attenuation of hepatic injury. Isolated hepatocytes were transfected with small interfering RNA to suppress GPR30 or ER. We found that suppression of GPR30 but not ER-alpha prevented E2-BSA- or E2-induced PKA activation and Bcl-2 expression. These results suggest that the nongenomic salutary effect of E2 in reducing hepatic injury after trauma-hemorrhage is mediated through the PKA-dependent pathway via GPR30 but not ER-alpha.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Animals
  • Blotting, Western
  • Cyclic AMP-Dependent Protein Kinases / antagonists & inhibitors
  • Cyclic AMP-Dependent Protein Kinases / metabolism*
  • Estradiol / administration & dosage
  • Estradiol / pharmacology*
  • Estradiol / therapeutic use
  • Estrogen Receptor alpha / genetics
  • Estrogen Receptor alpha / metabolism
  • Estrogens, Conjugated (USP) / administration & dosage
  • Estrogens, Conjugated (USP) / pharmacology
  • Estrogens, Conjugated (USP) / therapeutic use
  • Glutathione Transferase / blood
  • Hemorrhage / blood
  • Hemorrhage / etiology
  • Hemorrhage / prevention & control*
  • Hepatocytes / drug effects
  • Hepatocytes / metabolism
  • Isoenzymes / blood
  • Isoquinolines / administration & dosage
  • Isoquinolines / pharmacology
  • Liver / drug effects*
  • Liver / injuries
  • Liver / metabolism
  • Male
  • Models, Biological
  • Protein Kinase Inhibitors / pharmacology
  • Proto-Oncogene Proteins c-bcl-2 / metabolism
  • RNA, Small Interfering / genetics
  • Rats
  • Rats, Sprague-Dawley
  • Receptors, G-Protein-Coupled / genetics
  • Receptors, G-Protein-Coupled / metabolism
  • Receptors, G-Protein-Coupled / physiology*
  • Serum Albumin, Bovine / administration & dosage
  • Serum Albumin, Bovine / pharmacology*
  • Serum Albumin, Bovine / therapeutic use
  • Signal Transduction / drug effects
  • Sulfonamides / administration & dosage
  • Sulfonamides / pharmacology
  • Transfection
  • Wounds and Injuries / complications

Substances

  • Estrogen Receptor alpha
  • Estrogens, Conjugated (USP)
  • Isoenzymes
  • Isoquinolines
  • Protein Kinase Inhibitors
  • Proto-Oncogene Proteins c-bcl-2
  • RNA, Small Interfering
  • Receptors, G-Protein-Coupled
  • Sulfonamides
  • estradiol-bovine serum albumin
  • Serum Albumin, Bovine
  • Estradiol
  • Glutathione Transferase
  • glutathione S-transferase alpha
  • Cyclic AMP-Dependent Protein Kinases
  • N-(2-(4-bromocinnamylamino)ethyl)-5-isoquinolinesulfonamide