Troglitazone acts by PPARγ and PPARγ-independent pathways on LLC-PK₁-F⁺ acid-base metabolism

Troglitazone was studied in pH-sensitive LLC-PK₁-F⁺ cells to determine the effect on pH_i and glutamine metabolism as well as the role of peroxisome proliferator-activated receptor (PPARγ)-dependent and PPARγ-independent signaling pathways. Troglitazone induces a dose-dependent cellular acidosis that occurs within 4 min and persists over 18 h as a result of inhibiting Na⁺/H ⁺ exchanger-mediated acid extrusion. Cellular acidosis was associated with glutamine-dependent augmented [¹⁵N]ammonium production and decreased [¹⁵N]alanine formation from ¹⁵N-labeled glutamine. The shift in glutamine metabolism from alanine to ammoniagenesis appears within 3 h and is associated after 18 h with both a reduction in assayable alanine aminotransferase (ALT) activity as well as cellular acidosis. The relative contribution of troglitazone-induced cellular acidosis vs. the decrease in assayable ALT activity to alanine production could be demonstrated. The PPARγ antagonist bisphenol A diglycide ether (BADGE) reversed both the troglitazone-induced cellular acidosis and ammoniagenesis but enhanced the troglitazone reduction of assayable ALT activity; BADGE also blocked troglitazone induction of peroxisome proliferator response element-driven firefly luciferase activity. The protein kinase C (PKC) inhibitor chelerythrine mimics troglitazone effects, whereas phorbol ester reverses the effects on ammoniagenesis consistent with troglitazone negatively regulating the DAG/PKC/ERK pathway. Although functional PPARγ signaling occurs in this cell line, the major troglitazone-induced acid-base responses appear to be mediated by pathway(s) involving PKC/ERK.