Carlson, Ting

The effect of 25-hydroxycholesterol (25-OH-cholesterol) and chenodeoxycholic (CDC) acid on apoprotein secretion, low-density lipoprotein receptor activity, and [3H]triacylglycerol secretion in Hep G2 cells was studied. Both 25-OH-cholesterol and CDC acid increased the secretion of apolipoprotein (apo) E by Hep G2 cells. The secretion of apo A-I was slightly lowered (less than 10% disease). The maximal increase in apo E secretion was observed in culture medium containing 2 micrograms of 25-OH-cholesterol/ml or 10 micrograms of CDC acid/ml plus 10% fetal calf serum. Cholesterol, 7-OH-cholesterol and other bile acids were ineffective in inducing increases in apo E secretion. Another cholesterol synthesis inhibitor, mevinolin, was also ineffective in generating an increase in apoprotein secretion. The data indicated a specific interaction between 25-OH-cholesterol or CDC acid and apo E secretion in Hep G2 cells. Cholesterol synthesis, as measured by the incorporation of [14C]acetic acid into sterols, was repressed in Hep G2 cells in the presence of 25-OH-cholesterol (17% of control value). CDC acid, on the other hand, increased [14C]acetic acid incorporation (156% of control value). The number of LDL receptors in Hep G2 cells was decreased after incubation with 25-OH-cholesterol (62% of control value), but increased significantly after incubation with CDC acid (149% of control value). The secretion of [3H]triacylglycerol by Hep G2 cells incubated with 25-OH-cholesterol was greatly increased (248% of control value). On the contrary, CDC acid did not cause any increase in [3H]triacylglycerol secretion. The above results suggest that 25-OH-cholesterol and CDC acid have different effects on lipid metabolism in Hep G2 cells. The mRNA levels of apo E increased in cells preincubated with 25-OH-cholesterol and CDC acid, which suggested that the increase in apo E secretion is at least partly due to an increase in synthesis.

AbstractLipid and apolipoprotein (apo) A‐I concentrations in different density fractions of New Zealand White (NZW) and Watanabe (WHHL) rabbit plasma were studied. Aside from the low plasma apoA‐I and high density lipoprotein (HDL) cholesterol levels in WHHL rabbits, the distribution of apoA‐I was also different between the two rabbits. ApoA‐I was concentrated in both the HDL2 and HDL3 fractions of NZW rabbits but was found primarily in the HDL3 fraction of WHHL rabbits. ApoA‐I secretion in these two rabbits was further studiedin vitro by using intestinal and hepatocyte cell cultures. ApoA‐I secretion was highest from cultures of the duodenum and the proximal end of the jejunum; whereas, cell cultures of the distal end of the small intestine secreted very little apoA‐I into the medium. Intestinal cell cultures from WHHL rabbits secreted less, but significant amounts of, apoA‐I compared to that of NZW rabbits. ApoA‐I was most concentrated in the density range of 1.12–1.21 (HDL3) fraction in medium containing 10% fetal calf serum (FCS). Serum‐free medium promoted apoA‐I secretion by intestinal cell cultures that was mostly found in the d>1.21 (lipoprotein‐deficient) fraction. Hepatocytes isolated from the same rabbits by collagenase perfusion secreted little apoA‐I, and it was found only in the d>1.21 fraction. The addition of oleic acid into the culture medium with 10% FCS decreased the secretion of total apoA‐I and HDL by intestinal cell cultures and increased the secretion of very low density lipoprotein (VLDL) and intermediate density lipoproteins (IDL). The results indicate that intestinal cells, not hepatocytes, are responsible for the secretion of apoA‐I and HDL3 in rabbits, and that the secretion may be regulated under different nutritional conditions.