The use of High Performance Thin-Layer Chromatography to determine the role of membrane lipid composition in bile salt-induced kidney cell damage

Abstract

Introduction: In obstructive liver disease bile salts are known to accumulate in and damage specific kidney cells. High Performance Thin-Layer Chromatography (HPTLC) was used to determine the membrane lipid composition of a range of kidney cells. Methods: Kidney cells were exposed to three hydrophobic bile salts (lithocholic, deoxycholic and chenodeoxycholic acids) and cytotoxicity was determined. In addition membrane lipids from the cells were extracted in a chloroform:methanol (2:1, v/v) solution and quantified by HPTLC. Results: The results reveal a differential toxicity to the bile acids with IC 50 values ranging from 79 ± 5 μM to 394 ± 13 μM. When the lipid composition of the most and least susceptible cells were assayed, the least susceptible cells had a much higher lipid composition (46.6 ± 3.7 μg/mg protein compared to 28.1 ± 5.2 μg/mg protein for the least susceptible cells). Discussion: These results suggest that HPTLC may be a useful technique when determining the mechanisms of toxicity of compounds which cause the disruption of the cell membrane.