Role of adenine nucleotides in the activation of microsomal cholesterol ester hydrolase by fructose or adenosine in rat hepatocytes.
In the present study we have analysed the potential relationship between the cellular level of adenine nucleotides and the activity of microsomal cholesterol ester hydrolase by treating rat hepatocyte suspensions with fructose or adenosine. Fructose raised the microsomal hydrolysis of cholesteryl esters as a function of the dose. This ketose led to marked decreases in the cell level of ADP, ATP and total adenine nucleotide whereas that of AMP increased slightly, thus giving a rise in the cellular AMP/ATP ratio. The effects remained virtually constant over a period of 60 min. Incubation of hepatocytes in a Ca(2+)-free medium with or without ethylene glycol-bis(beta-aminoethyl ether) N,N,N',N'-tetraacetic acid blocked by 40% the fructose-induced activation of cholesterol esterase whereas the rise in AMP/ATP was unaffected. Adenosine caused dose-dependent activations of cholesterol ester hydrolase and raised AMP, ADP and ATP concentrations as well as the AMP/ATP ratio. 2-Chloro-adenosine and N6-[L-2-phenyl-isopropyl] adenosine, non-metabolizable analogues of adenosine, did not mimic the effects of the nucleoside. A positive linear correlation exists between the percentage rises in the activity of microsomal cholesterol ester hydrolase and those in the intracellular AMP/ATP ratio in fructose- or adenosine-treated cells. These results indicate that, in microsomes from intact hepatocytes, the breakdown of cholesteryl esters to yield cholesterol and fatty acids is stimulated by fructose and adenosine and this can be explained in part by the increase in the cellular AMP/ATP ratio. In the case of fructose, also a Ca(2+)-dependent mechanism is involved.
Hernandez
M L
ML
Department of Physiology, University of the Basque Country Medical School, Bilbao, Spain.
Martinez
M J
MJ
Ochoa
B
B
D000659:AMP Deaminase / Q000037:antagonists & inhibitors; D000225:Adenine / Q000031:analogs & derivatives; D000227:Adenine Nucleotides / Q000378:metabolism*; D000241:Adenosine / Q000494:pharmacology*; D000249:Adenosine Monophosphate / Q000378:metabolism; D000255:Adenosine Triphosphate / Q000378:metabolism; D000818:Animals; D002478:Cells, Cultured; D004789:Enzyme Activation; D005260:Female; D005632:Fructose / Q000494:pharmacology*; D008862:Microsomes, Liver / Q000187:drug effects / Q000201:enzymology*; D051381:Rats; D017207:Rats, Sprague-Dawley; D002787:Sterol Esterase / Q000378:metabolism*
D000227:Adenine Nucleotides; D005632:Fructose; D000249:Adenosine Monophosphate; C010192:9-(2-hydroxy-3-nonyl)adenine; D000255:Adenosine Triphosphate; D002787:Sterol Esterase; D000659:AMP Deaminase; D000225:Adenine; D000241:Adenosine