Evidence for the induction of fatty acid desaturation in proliferating hepatic endoplasmic reticulum in response to treatment with polychlorinated biphenyls. Are fatty acid desaturases cytochrome P-450-dependent monooxygenases?

1. Polychlorinated biphenyls (PCBs) are abundant and persistent pollutants in the ecosystem which accumulate in biological systems. 2. We have shown previously (Borlakoglu et al., 1990; Eur. J. Biochem. 118, 327-332) that 120 hr after treating pigeons and rats with 1.5 mmol Aroclor 1254/kg body weight, hepatic microsomal membranes showed significant increases in the proportion of arachidonate (20:4,5, 8,11,14), in the concentration of cytochrome P-450 and in the activities of a wide range of cytochrome P-450-dependent enzymes involved in the metabolism of drugs and other xenobiotics. 3. After treating pigeons and rats in vivo with Aroclor 1254, linoleate desaturases activity increased significantly 3.35-, 4.35-, 5.83- and 8.61-fold 24, 48, 68 and 120 hr for pigeons and 2- and 7-fold for rats respectively 48 and 120 hr post treatment. The total activity of linoleate desaturases in the whole liver of pigeons and rats increased 40- and 10-fold respectively. 4. There were excellent correlations between the concentrations of cytochrome b5 and cytochrome P-450 and the activity of pigeon linoleate desaturases. Extrapolation of the concentration of cytochrome P-450 to zero is coincident with zero linoleate desaturase activity. 5. Evidence is presented to suggest the novel concept that linoleate desaturation is dependent upon the catalytic cycle of these monooxygenases.

Transport and cellular uptake of polychlorinated biphenyls (PCBs)--II. Changes in vivo in plasma lipoproteins and proteins of pigeons in response to PCBs, and a proposed model for the transport and cellular uptake of PCBs.

The complex distribution of polychlorinated biphenyl (PCB) isomers and congeners amongst plasma fractions of the pigeon suggests that the lipid and apolipoprotein components of lipoproteins, as well as plasma proteins, may be important in transporting PCBs to tissues (Borlakoglu et al., Biochem. Pharmac. 40, 265 (1990]. Pigeons were injected with the commercial PCB mixture Aroclor 1254 (1.5 mmol/kg body weight). After 120 hr triacylglycerol-like droplets accumulated in hepatocytes ('fatty liver syndrome'), there was proliferation of the hepatic smooth endoplasmic reticulum, and plasma concentrations of triacylglycerol and total cholesterol increased. This was accompanied by significant decreases in plasma concentrations of total protein, total apolipoproteins of the low density lipoprotein (LDL) and the high density lipoprotein (HDL) fractions, and albumin and by a significant increase in that of urea, indicating increased protein breakdown. These results suggest that Aroclor 1254 increased hepatic lipid synthesis, but decreased hepatic production of albumin and apolipoproteins. This would explain the accumulation of triacylglycerol in the liver and the increase in the proportion of triacylglycerol to apolipoprotein in the total lipoproteins. From the evidence presented, a model is proposed based on the association of PCBs with hydrophobic domains of lipids and proteins for the transport of PCBs by plasma fractions, their uptake into cells and intracellular metabolism, and their accumulation in adipose tissue.

Polychlorinated biphenyls increase fatty acid desaturation in the proliferating endoplasmic reticulum of pigeon and rat livers.

1. Polychlorinated biphenyls (PCB) are abundant and persistent pollutants in the ecosystem. Commercial mixtures (e.g. Aroclor 1254) can contain up to 80 different isomers and congeners, many of which accumulate in biological systems by the ingestion of PCB-contaminated lipid components of food chains. 2. Commercial mixtures of PCB induce, in hepatic microsomal membranes in vivo, a variety of different forms of the cytochrome P-450 components of enzyme systems involved in the metabolism of drugs and other xenobiotics, and can also induce the proliferation of this membrane. Since these microsomal enzyme systems share a number of the requirements of microsomal fatty acid desaturases, we have investigated whether the induction by PCB in vivo of cytochrome-P-450-linked enzymes in the proliferating hepatic microsomal membrane of the pigeon and the rat is accompanied by increased proportions of polyunsaturated fatty acids in this membrane. 3. The most striking changes observed 120 h after treating pigeons and rats with 1.5 mmol Aroclor 1254/kg body mass were 2.2-fold and 1.6-fold increases, respectively, in the proportion of arachidonic acid in the hepatic microsomal membrane. When the effects of this treatment on the proliferation of this membrane and increase in liver mass are taken into account, the amount of arachidonic acid in the total microsomal membrane of pigeon and rat livers increased 6.7-fold and 1.9-fold, respectively. 4. These changes were accompanied by very significant increases in pigeons and rats of the concentration of hepatic microsomal cytochrome P-450, and in the activity in microsomal protein of a wide range of cytochrome P-450-dependent enzyme involved in the metabolism of drugs and other xenobiotics. 5. This effect of PCB, of increasing in vivo the degree of unsaturation of fatty acids of hepatic microsomal membrane, appears to be a novel finding, and does not seem to have been investigated for other drugs and xenobiotics. Preliminary results have shown that the effect is accompanied by substantial increases in the total activity of delta 6 and delta 5 microsomal fatty acid desaturases converting 18:2 (9, 12) (linoleic acid) to 20:4 (5, 8, 11, 14) (arachidonic acid) [Borlakoglu, J.T., Dils, R.R., Edwards-Webb, J.D. & Walker, C.H. (1988) Biochem. Soc. Trans. 16, 1072]. 6. It is postulated that there is a significant link between increased fatty acid desaturation and the induction of cytochrome-P-450-linked enzymes, and this is discussed in terms of the mechanisms involved in the metabolism of foreign compounds.

Evidence for the induction of cytochrome P-452 in rat liver by Aroclor 1254, a commercial mixture of polychlorinated biphenyls.

We have examined the ability of a commercial mixture of polychlorinated biphenyls (Aroclor 1254) to induce hepatic cytochrome P-452-linked enzyme activities in rat and pigeon liver five days after its intraperitoneal injection. The results provide evidence that, at the doses used, Aroclor 1254 induces cytochrome P-452-linked enzyme activities in rats, but not in pigeons. This inductive effect was previously regarded as being specific for hypolipidemic drugs and phthalate ester plasticisers.

Concurrent studies of the flow of digesta in the duodenum and of exocrine pancreatic secretion of calves. 7. Influence of milk substitutes on abomasal lipolysis and biliary secretion.

1. The abomasal hydrolysis of lipids and the flow of endogenous (biliary) lipids was studied in two Friesian calves given four milk-substitute diets, by sampling the duodenal digesta. The diets were: reconstituted, mildly preheated, spray-dried skim-milk powder with (SKF) or without (SK) margarine fat or with 500 g/kg skim-milk powder in diet SKF replaced by soy-beran flour (ASKF) or fish-protein concentrate (BSKF) together with dried whey. The diets were given ad lib. twice daily from 13 to 37 d of age, each diet being given for six consecutive days. Collections of duodenal digesta from the re-entrant cannula situated caudal to the bile duct were made for 12 h after feeding the 6th and 12th meals for each diet. Samples from one collection only were subjected to detailed analysis of the lipid classes. 2. The inclusion of non-milk protein (ASKF and BSKF) not only increased the rate of passage of lipid through the abomasum but also the proportion of the lipid present as triglyceride particularly, in the first 2 h after feeding. 3. In a 12 h period, 2.3-6.3 g 'polar' lipids (mainly biliary phospholipids) were estimated to have been secreted. The rate of flow was high during the first hour after feeding and constant thereafter. The quantity of 'polar' lipid was not related to the type of milk fed or the duodenal flow of lipid. 4. When diet SK was fed, the small amounts of lipid present were extensively hydrolysed so that free fatty acids represented 700 g/kg lipid of dietary origin passing through the duodenum. When margarine fat was included in the diets (SKF, ASKF and BSKF), the free fatty acids represented only 210 g/kg lipid of dietary origin. 5. The quantities of lipid and nitrogen passing through the duodenum were poorly related to the quantities ingested at the beginning of the 12 h experimental period but were closely related to each other.

Studies on lipid digestion in the preruminant calf. 2. A comparison of the products of lipolysis of milk fat by salivary and pancreatic lipases in vitro.

1. A comparison has been made of the products of lipolysis of the fat in cow's milk by salivary and pancreatic lipases (EC 3-1-1-3) from a preruminant calf. 2. It was confirmed that salivary lipase releases short-chain acids preferentially, particularly butyric and caproic acids. This was not only true initially but also as the course of lipolysis progressed. Even after 60 min incubation the longer-chain acids were only released to about one-twentieth of the extent of butyric acid. 3. Although pancreatic lipase also initially released proportionately more butyric acid than other fatty acids, within 30 min the longer-chain acids were released to about half the extent of butyric acid. 4. Since the over-all extent of lipolysis by salivary lipase was limited by its inability to release long-chain acids, the release of these acids (which comprise two-thirds of the total fatty acids of milk fat) in the calf is likely to be dependent on the subsequent action of pancreatic lipase. 5. The release of fatty acids by pancreatic lipase was enhanced by pre-incubating milk fat with salivary lipase.

Studies on lipid digestion in the preruminant calf. The source of lipolytic activity in the abomasum.

1. Lipolytic and proteolytic activities and pH values were determined in secretions collected from innervated abomasal pouches and in abomasal contents from preruminant calves given liquid diets. 2. No lipolytic activity was detected in pouch secretions collected during 1 h after feeding, though lipolytic activity was present in abomasal contents; pepsin (EC 3.4.23.1) and renin (EC 3.4.23.4) were present in both pouch secretions and abomasal contents. The pH values of pouch secretions ranged from 1-2 to 1-8 and those of abomasal contents from 4-2 to 5-9. 3. When diet was placed directly into the abomasal pouch soon after feeding, the pH values of pouch and abomasal contents decreased similarly (i.e. from 6-3 to approximately 5). Protease activity (U/ml) of pouch contents ranged from 0-1 to 0-8 and that of abomasal contents from 0-1 to 0-2. No lipolytic activity was detected in pouch contents, though abomasal contents contained 0-6 to 1-2 U/ml and when the diet contained milk-fat as the dietary fat source considerable lipolysis of triglycerides containing shorter-chain fatty acids was found. 4. It is concluded that there is no significant secretion of lipolytic enzymes by the fundal mucosa and that the lipolysis of triglycerides in the abomasum of the preruminant calf is due predominantly to a lipolytic enzyme in saliva.

Studies on metabolism of vitamin A. The effect of vitamin A status on the secretion rates of some steroids into the ovarian venous blood of pregnant rats.

1. Rats raised on a vitamin A-deficient diet supplemented with either retinyl acetate or retinoic acid were mated and became pregnant. 2. The rates of secretion of progesterone, 20alpha-hydroxypregn-4-en-3-one, oestradiol-17beta and oestrone into the ovarian-venous blood of rats in these two groups were measured on days 9 and 15 of pregnancy. 3. Rates of secretion of progesterone and 20alpha-hydroxypregn-4-en-3-one, both on days 9 and 15, were lower for the rats given retinoic acid. No such differences were found in ovarian oestrogen secretion. 4. The implications of these results are discussed in the light of the previous demonstration that the activity of ovarian 3beta-hydroxy-Delta(5)-steroid dehydrogenase was markedly less in pregnant rats given retinoic acid.