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TOXIC INTERACTIONS BETWEEN CARBON TETRACHLORIDE AND CHLOROFORM IN CULTURED RAT HEPATOCYTES
Citation:
Lamb, R., J. Borzelleca, L. Condie, AND C. Gennings. TOXIC INTERACTIONS BETWEEN CARBON TETRACHLORIDE AND CHLOROFORM IN CULTURED RAT HEPATOCYTES. U.S. Environmental Protection Agency, Washington, D.C., EPA/600/J-89/330.
Description:
Primary cultures of adult rat hepatocytes were incubated (1.5-16 hr) with various concentrations of CC14 (<0.5 mM) and/or CHCl3 (<2.5 mM). gent dependent alterations in hepatocyte functions were assessed by measuring (1) [3H]choline incorporation into phosphatidylcholine (endoplasmic reticulum), (2) MTT (tetrazolium salt) reduction (mitochondria) and (3) AST release into medium (plasma membrane). ultured hepatocytes incubated with 0.5 mM CCl4 displayed a significant (p < 0.001) and rapid (1.5 hr) reduction (40%) in endoplasmic reticulum function that preceded significant (p < 0.001) alterations in mitochondria (6-16 hr) and plasma membrane (6-16 hr) functions. C14-dependent alterations in liver cell functions are a result of CCl4 bioactivation since metyrapone inhibits the CCl4 mediated changes in cell functions. esponse surface methods (RSM) were used to determine the influence of combinations of CCl4, and CHCl3 on liver cell MTT reduction and [3H]choline incorporation. egression coeffients were determined for CCl4, CHCl3, and CCl4-CHCl3. ll results were significant (p < 0.0001) and implied that CCl4 was a more potent hepatotoxin in vitro than CHCl3. he RSM analysis also suggested that combinations of CHCl3 and CCl4 have greater than additive effects on MTT reduction and [3H]choline incorporation. hese effects of CCl4 and/or CHCl3 on liver cell functions in vitro are consistent with liver alterations observed in vivo. herefore, primary cultures of adult rat hepatocytes may be an appropriate model in vitro to the hepatotoxic potential of agents alone or in combination.