The use of High Performance Thin-Layer Chromatography to determine the role of membrane lipid composition in bile salt-induced kidney cell damage.
BACKGROUND
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 microM to 394+/-13 microM. 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 microg/mg protein compared to 28.1+/-5.2 microg/mg protein for the least susceptible cells).


CONCLUSIONS
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.
Morgan
Winston A
WA
School of Health and Bioscience, University of East London, Romford Road, London E15 4LZ, United Kingdom. w.a.morgan@uel.ac.uk
Nk
Thanh
T
Ding
Yinuo
Y
D000818:Animals; D002417:Cattle; D002460:Cell Line; D002470:Cell Survival / Q000187:drug effects; D002635:Chenodeoxycholic Acid / Q000494:pharmacology*; D002855:Chromatography, Thin Layer; D003840:Deoxycholic Acid / Q000494:pharmacology*; D004285:Dogs; D005562:Formazans / Q000494:pharmacology; D007668:Kidney / Q000166:cytology* / Q000187:drug effects*; D008095:Lithocholic Acid / Q000494:pharmacology*; D008563:Membrane Lipids / Q000378:metabolism*; D009893:Opossums; D051381:Rats; D013778:Tetrazolium Salts / Q000494:pharmacology
D005562:Formazans; D008563:Membrane Lipids; D013778:Tetrazolium Salts; D003840:Deoxycholic Acid; D002635:Chenodeoxycholic Acid; C079782:MTT formazan; D008095:Lithocholic Acid