In rat livers perfused with undiluted rat blood at perfusion rates of 6, 12, or 18 ml/min, hepatic O2 consumption rose with blood flow. Lipogenesis was unaffected by blood flow in control livers and was enhanced by insulin at 12 and 18 ml/min. Very-low-density lipoprotein triacylglycerol secretion also rose with increased flow and was stimulated by insulin at both 6 and 12 ml/min. When glucose was added to livers perfused at 12 or 18 ml/min, uptake was independent of perfusion rate and was slightly stimulated by insulin. Total lipogenesis and the secretion of newly synthesized fatty acids in very-low-density lipoprotein triacylglycerols were unaffected by insulin at either flow rate. The hormone stimulated triacylglycerol secretion at 18 ml/min but inhibited it at 12 ml/min. It seems that in perfused liver, effects of insulin on lipogenesis and very-low-density lipoprotein secretion may be modified not only by changes in O2 consumption (in this case through alterations in blood flow) but also by the choice of substrate.