GENTAMICIN-INDUCED ALTERATIONS IN PIG-KIDNEY EPITHELIAL (LLC-PK1) CELLS IN CULTURE

Google Scholar
  • 1 January 1986
    • journal article
    • research article
    • Vol. 236  (1) , 254-262
Abstract
The effect of gentamicin exposure was investigated in LLC-PK1 cells in culture. Gentamicin (0.5-2.0 mM) was added to the medium of cells which had been grown to confluency in the absence of antibiotics and antimycotics. Exposure to gentamicin (1-4 days) did not effect total cellular protein or DNA levels, total cell number or the release of various marker enzymes to the medium. ATP levels in gentamicin-treated cells did not differ from control cells; however, medium from the gentamicin-treated cells contained significantly lower lactic acid levels. Morphological examination by electron microscopy revealed gentamicin-elicited myeloid body formation. Furthermore, total phospholipid level was elevated markedly in gentamicin-treated cells. Analysis of specific phospholipid classes showed only phosphatidylcholine, phosphatidylinositol and polyphosphoinositide phospholipid levels increased in a time-dependent manner. Phosphatidylinositol showed the highest percentage of increase. Raising the normal medium calcium concentration (0.2 mg/ml) 1.5-, 2.0- or 3.0-fold did not alter gentamicin-induced elevation in cellular phosphatidylinositol and phosphatidylcholine. Gentamicin exposure also resulted in a concentration-dependent increase in the turnover of LLC-PK1 cell-free fatty acids, monoglyceride, diglyceride and nonesterified cholesterol and decrease in triglyceride turnover. Calcium transport into and through the cell monolayer was inhibited markedly by gentamicin despite the fact that 45Ca++ binding to gentamicin-treated cells was greater. These results demonstrate that manifestations of gentamicin toxicity in LLC-PK1 cells parallel those reported in the whole animal thus making the LLC-PK1 cell in culture a valid system for elucidating the mechanism of gentamicin-elicited alterations in renal epithelium.