Mechanism of aminoglycoside-induced lysosomal phospholipidosis: in vitro and in vivo studies with gentamicin and amikacin.
Gentamicin, a widely used aminoglycoside antibiotic, is concentrated in lysosomes of proximal tubular cells of the kidney, and induces therein an accumulation of myelin-like material. We show that treatment of rats with Gentamicin (10 mg/kg, 7 days) induces a loss of activity of lysosomal sphingomyelinase and phospholipase A1, associated with an increase in the amount of total lipid phosphorus in the kidney cortex. In vitro, Gentamicin is shown by gel permeation to bind to phospholipid bilayers (liposomes) under conditions which mimic the lysosomal environment (acid pH and presence of phosphatidylinositol). The reversal of this binding by an increase in the ionic strength (less than 0.04) suggests electrostatic interaction between the hydrophilic, polycationic aminoglycoside and the negatively charged phospholipids. Binding of Gentamicin impairs the hydrolysis of phosphatidylcholine present in the bilayer, by lysosomal phospholipases A1 and A2 from the liver or kidney. We also show that lysosomal sphingomyelinase is readily and irreversibly inactivated by liposomes in the absence of detergent. The lysosomal phospholipidosis induced by Gentamicin in the kidney, as in cultured cells [Aubert-Tulkens et al., Lab. Invest. 40, 481 (1979)] appears therefore to be a direct consequence of the lysosomotropic character of this drug and its ability to inhibit therein phospholipid breakdown. Amikacin, a semi-synthetic aminoglycoside, binds more loosely to phospholipid bilayers, induces less inhibition of phospholipases in vitro and is less taken up by tubular cells in vivo. Accordingly, Amikacin does not provoke significant lysosomal phospholipidosis or loss of sphingomyelinase and phospholipase A1 activities in vivo at the doses and time investigated (0-40 mg/kg, 7 days). Inasmuch as Amikacin is reported to be less toxic to the kidney, we suggest that lysosomal alterations are an early and significant step in aminoglycoside-induced nephrotoxicity.
D000583:Amikacin / Q000494:pharmacology*; D000818:Animals; D005260:Female; D005839:Gentamicins / Q000494:pharmacology*; D007612:Kanamycin / Q000031:analogs & derivatives*; D008247:Lysosomes / Q000378:metabolism*; D010743:Phospholipids / Q000378:metabolism*; D051381:Rats; D011919:Rats, Inbred Strains; D013109:Sphingomyelins / Q000378:metabolism
D005839:Gentamicins; D010743:Phospholipids; D013109:Sphingomyelins; D007612:Kanamycin; D000583:Amikacin