How to obtain cytosolic fraction from liver homogenate?
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We have observed, as have others, that liver homogenates from healthy rats are capable of inactivating small (microgram) quantities of endotoxins. In contrast, if the rats received lead acetate intravenously prior to the preparation of liver homogenates, no endotoxin inactivation was observed. Similar results were obtained if we incubated natural bile fluid with lead acetate. Such bile solutions lost their capacity to detoxify endotoxin. It is likely that lead acetate damages the side chains that play an important role in the detergent effect of bile acids, and for this reason the modified bile acids are unable to detoxify endotoxin. This phenomenon may play a role in the lead acetate induced endotoxin sensitivity in vivo.
We demonstrated that after the dissociation of endotoxin by bile acids to its basic units, the detoxified material maintains the capacity to induce endotoxin tolerance. It is possible that this observation will lead to the isolation of non-toxic endotoxin subunits, which will be able to stimulate natural resistance without toxic side effects.
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