Apical ANG II-stimulated PLA2 activity and Na+ flux: a potential role for Ca2+-independent PLA2

Am J Physiol. 1997 Oct;273(4):F554-62. doi: 10.1152/ajprenal.1997.273.4.F554.

Abstract

Type 1 angiotensin II (ANG II) receptors (AT1R), which mediate proximal tubule (PT) salt and water reabsorption, undergo endocytosis and recycling. Prior studies in a PT-like model (LLC-PKcl4 cells expressing rabbit AT1R) (LLC-PK-AT1R cells) determined that quinacrine, a nonspecific phospholipase A2 (PLA2) inhibitor, and the haloenol lactone suicide substrate (HELSS), a Ca2+-independent PLA2 inhibitor, attenuated apical (AP) AT1R recycling. Further studies were undertaken to examine the association between AT1R endocytotic movement and PLA2 activity in this model. AP ANG II (100 nM) increased [3H]arachidonic acid ([3H]AA) release 4.4 +/- 0.38-fold in LLC-PK-AT1R cells cultured on permeable supports. Basolateral (BL) ANG II had no significant effect. Reversed-phase high-performance liquid chromatography confirmed that AP ANG II stimulated free [3H]AA release. Quinacrine, HELSS, and palmitoyl trifluoromethyl ketone, another Ca2+-independent PLA2 inhibitor, inhibited AP ANG II-stimulated [3H]AA release, as did inhibiting AP AT1R internalization with phenylarsine oxide. The role of HELSS-inhibitable AA release in ANG II-mediated 22Na flux was examined, given the effects of AT1R-mediated PLA2 activity on salt and water reabsorption. AP ANG II (100 nM) stimulated 22Na flux (AP--> BL), a response inhibited by HELSS. Thus, in this model, AP AT1R activated PLA2 with concomitant 22Na flux (AP --> BL), suggesting a link between AP AT1R endocytotic movement, AT1R-stimulated PLA2 activity, and 22Na flux in this model. The effects of HELSS suggest that Ca2+-independent PLA2 activity may be involved in this AP ANG II response.

Publication types

  • Research Support, U.S. Gov't, Non-P.H.S.
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Angiotensin II / pharmacology*
  • Animals
  • Arachidonic Acid / metabolism
  • Calcium / physiology
  • Cell Membrane / metabolism
  • Clone Cells
  • LLC-PK1 Cells
  • Naphthalenes / pharmacology
  • Phosphodiesterase Inhibitors / pharmacology
  • Phospholipases A / metabolism*
  • Phospholipases A2
  • Pyrones / pharmacology
  • Rabbits
  • Receptors, Angiotensin / metabolism
  • Sodium / metabolism*
  • Swine

Substances

  • Naphthalenes
  • Phosphodiesterase Inhibitors
  • Pyrones
  • Receptors, Angiotensin
  • Angiotensin II
  • Arachidonic Acid
  • 6-(bromomethylene)tetrahydro-3-(1-naphthaleneyl)-2H-pyran-2-one
  • Sodium
  • Phospholipases A
  • Phospholipases A2
  • Calcium