Adenylate energy charge of rat and human cultured hepatocytes |
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Authors: | Yoichi Matsui Hiroaki Kitade Tomoo Kamiya Toshiki Kanemaki Yoshifumi Hiramatsu Tadayoshi Okumura Yasuo Kamiyama |
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Institution: | (1) First Department of Surgery, Kansai Medical University, 1 Fumizono, Moriguchi, 570 Osaka, Japan;(2) Department of Medical Chemistry, Kansai Medical University, 1 Fumizono, Moriguchi, 570 Osaka, Japan |
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Abstract: | Summary A simple and rapid method for the assay of adenine nucleotides (ATP, ADP, and AMP) was established to evaluate the adenylate
energy charge (ATP+ADP/2)/(ATP+ADP+AMP) of cultured hepatocytes. The effects of inhibitors of glycolysis, fatty acid oxidation,
or oxidative phosphorylation on the energy charge were examined. The energy charges of cultured hepatocytes in rats and human
were almost identical and were maintained at a high level between 6 and 24 h after changing the media (rat: 0.908±0.008n=9, human: 0.918±0.014n=6, mean ± SD). Inhibition of glycolysis with sodium fluoride or oxidative phosphorylation with antimycin A irreversibly reduced
both the adenine nucleotide contents and the energy charge. However, the inhibition of fatty acid oxidation with 2-tetradecylglycidic
acid did not affect the nucleotide contents, and the energy charge only decreased transiently to recover within 8 h. When
the inhibitor of oxidative phosphorylation was removed, the recovery in the energy charge preceded the recovery in the adenine
nucleotide contents. These findings suggest that the adenylate energy charge is a more sensitive measure of the changes in
energy metabolism than the adenine nucleotide contents. Furthermore, energy charge regulates adenine nucleotide contents in
cultured hepatocytes. It is important to confirm that the high energy charge of the cultured hepatocytes is maintained when
these cells are used for metabolic studies. |
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Keywords: | adenine nucleotides adenylate energy charge antimycin A sodium fluoride tetradecylglycidic acid hepatocyte culture |
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