Hyperglycemia via activation of thromboxane A2 receptor impairs the integrity and function of blood-brain barrier in microvascular endothelial cells

Oncotarget. 2017 May 2;8(18):30030-30038. doi: 10.18632/oncotarget.16273.

Abstract

Diabetes is one of high risk factors for cardio- and cerebra-vascular diseases, including stroke, atherosclerosis and hypertension. This study was conducted to elucidate whether and how thromboxane receptor (TPr) activation contributes to blood-brain barrier (BBB) dysfunction in diabetes. Human brain microvascular endothelial cells (HBMECs) were cultured. The levels of phosphorylated endothelial nitric oxide synthase (eNOS) at Ser1177 (p-eNOS) and Akt at Ser473 (p-Akt) were assayed by western blot. Exposure of HBMECs to either high glucose (HG) or thromboxane A2 (TxA2) mimetic U46619, significantly reduced p-eNOS and p-Akt. These effects were abolished by pharmacological or genetic inhibitors of TPr. HG/U46619-induced suppressions of eNOS and Akt phosphorylation were accompanied by upregulation of PTEN and Ser380/Thr382/383 PTEN phosphorylation. PTEN-specific siRNA restored Akt-eNOS signaling in the face of TPr activation or HG. The small GTPase, Rho, was also activated by HG stimulation, and pretreatment of HBMECs with Y27632, a Rho-associated kinase (ROCK) inhibitor, rescued HG-impaired Akt-eNOS signaling. In STZ-injected rats, we found that hyperglycemia dramatically increased the levels of PTEN and PTEN-Ser380/Thr382/383 phosphorylation, reduced both levels of p-eNOS and p-Akt, and disrupted BBB function assayed by Evans blue staining, which were abolished by SQ29548 treatment. We conclude that hyperglycemia activates thromboxane A2 receptor to impair the integrity and function of blood-brain barrier via the ROCK-PTEN-Akt-eNOS pathway.

Keywords: blood-brain barrier; diabetes; thromboxane A2 receptor.

MeSH terms

  • Animals
  • Blood Glucose
  • Blood-Brain Barrier / metabolism*
  • Capillary Permeability
  • Endothelial Cells / metabolism*
  • Humans
  • Hyperglycemia / blood
  • Hyperglycemia / metabolism*
  • Male
  • Nitric Oxide Synthase Type III / metabolism
  • Phosphorylation
  • Proto-Oncogene Proteins c-akt / metabolism
  • Rats
  • Receptors, Thromboxane A2, Prostaglandin H2 / metabolism*
  • Signal Transduction
  • rho-Associated Kinases / metabolism
  • rhoA GTP-Binding Protein / metabolism

Substances

  • Blood Glucose
  • Receptors, Thromboxane A2, Prostaglandin H2
  • Nitric Oxide Synthase Type III
  • Proto-Oncogene Proteins c-akt
  • rho-Associated Kinases
  • rhoA GTP-Binding Protein