Leukotriene D4-induced proliferation of glomerular epithelial cells: PKC- and Na+-H+ exchanger-mediated response

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Abstract
The growth-promoting effect of leukotriene D4 (LTD4) has been observed in a variety of cells, including human glomerular epithelial cells. The purpose of this study was to determine the mechanisms underlying this process. LTD4 induction of [3H]thymidine uptake in human glomerular epithelial cells was blocked by the LTD4 receptor antagonist L648,051 when added in a 50-fold excess and by pertussis toxin. Neither drug affected basal DNA synthesis. These results suggest that the LTD4-mediated signal transduction implies activation of a GTP-binding protein that is coupled to a specific receptor. The possible role of protein kinase C (PKC) activation was also studied. In the presence of the PKC inhibitor H-7 or after downregulation of PKC levels by chronic treatment with phorbol ester, stimulation of [3H]thymidine uptake by LTD4 was greatly inhibited. Moreover, treatment of the cells by LTD4 resulted in a time-dependent increase of cytosolic PKC activity, whereas addition of phorbol 12-myristate 13-acetate reduced this activity. Therefore PKC-dependent mechanisms are likely to mediate the growth-promoting effect of LTD4. Finally, three approaches were used to determine the potential role of the Na+-H+ exchanger. First, progressive removal of extracellular Na+ using N-methyl-D-glucamine+ as a substitute inhibited LTD4-induced [3H]thymidine uptake with a 50% inhibitory concentration (IC50) of 85 mM. Second, addition of amiloride reduced the LTD4 growth effect with an IC50 of 6.5 microM, whereas three amiloride analogues exhibited lower IC50 values in accordance with their greater affinity for the Na+-H+ exchanger.(ABSTRACT TRUNCATED AT 250 WORDS)