Intracellular Angiotensin II Stimulation of Sodium Transporter Expression in Proximal Tubule Cells via AT(1) (AT(1a)) Receptor-Mediated, MAP Kinases ERK1/2- and NF-кB-Dependent Signaling Pathways

The current prevailing paradigm in the renin-angiotensin system dictates that most, if not all, biological, physiological, and pathological responses to its most potent peptide, angiotensin II (Ang II), are mediated by extracellular Ang II activating its cell surface receptors. Whether intracellular (or intracrine) Ang II and its receptors are involved remains incompletely understood. The present study tested the hypothesis that extracellular Ang II is taken up by the proximal tubules of the kidney by an AT(1) (AT(1a)) receptor-dependent mechanism and that overexpression of an intracellular Ang II fusion protein (ECFP/Ang II) in mouse proximal tubule cells (mPTC) stimulates the expression of Na(+)/H(+) exchanger 3 (NHE3), Na(+)/HCO(3)(-) cotransporter, and sodium and glucose cotransporter 2 (Sglt2) by AT(1a)/MAPK/ERK1/2/NF-kB signaling pathways. mPCT cells derived from male wild-type and type 1a Ang II receptor-deficient mice (Agtr1a(-/-)) were transfected with an intracellular enhanced cyan fluorescent protein-tagged Ang II fusion protein, ECFP/Ang II, and treated without or with AT(1) receptor blocker losartan, AT(2) receptor blocker PD123319, MEK1/MEK2 inhibitor U0126, NF-кB inhibitor RO 106-9920, or p38 MAP kinase inhibitor SB202196, respectively. In wild-type mPCT cells, the expression of ECFP/Ang II significantly increased NHE3, Na(+)/HCO(3)(-), and Sglt2 expression (p < 0.01). These responses were accompanied by >3-fold increases in the expression of phospho-ERK1/2 and the p65 subunit of NF-кB (p < 0.01). Losartan, U0126, or RO 106-9920 all significantly attenuated ECFP/Ang II-induced NHE3 and Na(+)/HCO(3)(-) expression (p < 0.01). Deletion of AT(1) (AT(1a)) receptors in mPCT cells attenuated ECFP/Ang II-induced NHE3 and Na(+)/HCO(3)(-) expression (p < 0.01). Interestingly, the AT(2) receptor blocker PD123319 also attenuated ECFP/Ang II-induced NHE3 and Na(+)/HCO(3)(-) expression (p < 0.01). These results suggest that, similar to extracellular Ang II, intracellular Ang II may also play an important role in Ang II receptor-mediated proximal tubule NHE3, Na(+)/HCO(3)(-), and Sglt2 expression by activation of AT(1a)/MAPK/ERK1/2/NF-kB signaling pathways.