16,16-Dimethyl PGE2 and fatty acids protect hepatocytes against CCl4-induced damage

Summary 16,16-Dimethyl PGE₂ (dmPGE₂) has previously been shown to protect the in vivo rat liver against CCl₄-induced damage. These studies were undertaken to determine if this protection could be demonstrated in vitro where factors of absorption, secretion, and blood flow are not present. Primary hepatocyte cultures were established by perfusing rat liver with collagenase. Hepatocytes were plated at a density of 2×10⁴ cells/cm, allowed 90 min to attach, then stabilized in L15 medium for 18 h. Hepatocytes were then challenged with CCl₄ with concomitant exposure to 10⁻⁹ to 10⁻⁵ M dmPGE₂, stearic acid, oleic acid, or ethanol vehicle (0.00001 to 0.1%). After 1 h, challenge was aspirated and cells were stained with 0.04% trypan blue to determine viability. Hepatocytes in the vehicle groups took up more trypan when exposed to CCl₄ than those treated with dmPGE₂, stearic acid, or oleic acid at concentrations of 10⁻⁹ to 10⁻⁷ M. At 0.1% ethanol vehicle protected as well as all other treatments. Protection against CCl₄ by dmPGE₂, stearic, and oleic acids as well as high concentrations of ethanol may occur by altering the metabolism of CCl₄.