Behavior of biliary phospholipids in intestinal lumen during fat digestion in rat

The phospholipids present in the intestinal lumen of rats following ingestion of triglycerides are of biliary origin. They consist of lecithins accompanied by a small proportion of lysolecithins. Their behavior in comparison with the other lipid constituents of the intestinal content was studied by subjecting the latter to gel filtration on an agarose column in the presence of a solution of 6 mM sodium taurocholate in 0.1 M NaCl. Part of the phospholipids is present with the triglycerides and diglycerides in the emulsified phase excluded from the gel where pancreatic lipase and colipase also are found. The remainder is found in optically clear fractions containing fatty acids, monoglycerides, and bile salts. These fractions are eluted at 2.0 column volumes, while mixed fatty acids, monoglycerides, bile salts micelles emerge from the column at 2.4 column volumes in the same chromatographic conditions. This difference in behavior may be explained by the presence of biliary lecithins. This presence could have an important bearing upon the mucosal uptake of the lipolysis products of triglycerides.     

Hydrolysis of acylglycerols and phospholipids of milled rice surface lipids during storage

The relative contribution of acylglycerols and phospholipids to the lipid hydrolysis in milled rice was investigated during storage at 37°C and 70% RH for 50 d. The MAG, DAG, and lysophospholipid contents of surface lipid were determined by reversed-phase HPLC. The MAG and DAG content of milled rice increased during storage from 0.06 to 0.18% (w/w milled rice), with the MAG content increasing more than that of the DAG. Lysophosphatidylcholine increased throughout the study from 0.013 to 0.034% (w/w), whereas lysophosphatidylinositol and lysophosphatidylethanolamine contents initially increased from 0.002 to 0.003% and from 0.005 to 0.009% (w/w), respectively, and then stabilized until day 50. The relative percentage of TAG and phospholipids hydrolyzed in milled rice increased rapidly during the first 3 d of storage from 12.3 to 37.6% and 25.0 to 62.5% (w/w), respectively, and steadily increased to 53.1 and 73.8% (w/w) on day 50. A higher percentage (62.5%) of phospholipids was hydrolyzed relative to TAG (37.6%) after 3 d and probably contributed significantly to milled rice lipid hydrolysis during early storage. However, TAG concentration (0.57%, w/w) was much higher relative to phospholipids (0.08%, w/w) in surface lipids, and therefore TAG hydrolysis was the major contributor to FFA development and the quality of stored milled rice.     

Homeostasis of mucosal cholesterol in the small intestine of the rat

This study was undertaken to investigate the capacity of the intestinal mucosa to maintain a constant cholesterol content under conditions where mucosal uptake or cholesterol transport into the lymph were manipulated. Two series of bile-diverted unanaesthetised rats were infused intraduodenally with saline, triolein emulsified with Pluronic F68, or taurocholate with or without added tomatine. Pluronic F68 is a nontoxic detergent which promotes mucosal uptake of polar lipids but not cholesterol. Tomatine is a cholesterol-binding saponin. One series of rats was used for measuring mucosal cholesterol content, DNA and protein after the test infusions. A second series of rats had the thoracic lymph duct cannulated but otherwise remained the same as the first series. The second series was used for measuring the effect of the different infusions on mass cholesterol output into lymph. Mucosal cholesterol content of rats that were not fed decreased with bile-diversion and was restored with taurocholate infusion. This suggested a contribution of luminal cholesterol to the mucosal cholesterol pool. However, evidence for a contribution from the lumen was provided by only one of two groups of rats given infusions which did not promote mucosal uptake of cholesterol. First, addition of tomatine to the taurocholate infusate prevented both the increase in lymph output of cholesterol and the increased mucosal cholesterol content shown in rats given taurocholate alone. Second, in another group of rats in which mucosal uptake of cholesterol was prevented, i.e. in rats given Pluronic F68-triolein emulsions, the increased fat absorption was accompanied by a marked increase in cholesterol output into lymph without a concomitant decrease in mucosal cholesterol content. These results would be consistent with increased mucosal synthesis of cholesterol as a possible source of endogenous cholesterol absorbed into lymph.     

The behavior of rat bile phospholipids in the intestine and in incubation media containing pancreatic juice

Samples of radioactive bile were collected from rats after intravenous injection of potassium soaps ([9–10³H₂] or [1¹⁴C] oleate, [1¹⁴C] linoleate or [9–10³H₂] palmitate). These radioactive acids were chosen because it is well established that, in natural phosphatidyl cholines, palmitic acid is located chiefly at the 1 position and linoleic and oleic acids at the 2 position. After incubation of bile with pancreatic juice, the labeling of unchanged biliary phospholipids was higher when native bile was labeled with oleic acid than with palmitic or linoleic acids. These data suggest that monounsaturated molecular species of biliary phospholipids are more resistant than the diunsaturated ones to in vitro hydrolysis by phospholipase A₂. Ninety min after introduction of the radioactive bile into the upper part of the rat duodenum, high labeling of luminal phospholipids was observed regardless of the bile sample used, although labeling of free fatty acids was always low. The passage of intact biliary phospholipids through the intestinal epithelium is discussed.     

Effects of dietary fish oil on biliary phospholipids and prostaglandin synthesis in the cholesterol-fed pairie dog

Cholesterol gallstone formation in the prairie dog is accompanied by an increase in the percentage of biliaryphospholipids containing arachidonic acid, and an increase in gallbladder prostaglandin (PG) synthesis, but the pathogenetic significance of these changes is unclear. Dietary supplementation with eicosapentaenoic acid (EPA), an omega-3 fatty acid which is commonly found in fish oil, decreases prostaglandin synthesis in some tissues by replacing arachidonic acid, and by competitively inhibiting prostaglandin synthesis. We studied the effect of dietary fish oil on gallbladder PG synthesis, and the relative abundance of various molecular species of phosphatidylcholines and phosphatidylethanolamines in bile and gallbladder epithelium in the cholesterol-fed prairie dog. Prairie dogs were maintained for 4 weeks on one of four diets: i) control, ii) cholesterol-supplemented (0.34%), iii) menhaden oil (50 g/kg chow), or iv) cholesterol plus menhaden oil. Supplementation with menhaden oil resulted in a replacement of arachidonic and linoleic acids with EPA and docosahexaenoic acids in the phospholipids of bile and gallbladder mucosa. In cholesterol-fed animals, supplementation with menhaden oil prevented increased gallbladder PG synthesis. Menhaden oil also reduced the incidence of cholesterol monohydrate crystals among cholesterol-fed animals (9/20 with cholesterol plus menhaden oil vs 21/22 with cholesterol alone), but the improvement could not clearly be attributed to decreased PG synthesis since supplementation with menhaden oil also increased the total phospholipid concentration in bile, and decreased the degree of cholesterol saturation. These results demonstrate that dietary supplementation with omega-3 fatty acids significantly influences biliary phospholipids, and decreases the incidence of cholesterol monohydrate crystal formation in this animal model.     

Structure determinations and synthesis of pharmacologically active phospholipids from kidney and intestine

The chemical structures of an acidic phospholipid originally isolated from equine intestine by Vogt and of a phospholipid isolated from rabbit kidney medulla by Walaszek are shown by total synthesis to be similar mixtures of 2-alkyl (and alkenyl)-4-hydroxymethyl-1,3-dioxolane-dihydrogen phosphate esters. In these materials, the alkyl residues derive primarily from oleyl, palmityl and linoleyl aldehydes. The smooth muscle contracting activity observed in the natural substances is shown to reside exclusively in the oleyl aldehyde derivative.     

Resistance to the effect of phospholipase A2 of the biliary phospholipids during incubation of bile

Mixtures of fresh bile of the rat and of isolated hepatic phospholipids (one or the other of these components having been labeled with³H oleic acid) were incubated either with heated rat pancreatic juice, at 37 C during periods of 1 and 3 hr, or with snake venom, at 25 C during periods of 17 and 36 hr, as sources of phospholipase A₂. After incubation, tritiated free oleic acid was measured since this acid was in the 2 position of both phospholipidic substrates. With heated pancreatic juice, no significant enzymatic hydrolysis of the bile phospholipids occured, but isolated hepatic phospholipids were readily attacked. With snake venom, the whole isolated hepatic phospholipids were very strongly hydrolyzed while biliary phospholipids were hydrolyzed to a much lesser extent.     

Digestion and assimilation features of dietary DAG in the rat small intestine

Several recent studies have demonstrated that dietary DAG oil rich in 1,3-species suppresses the postprandial increase of serum TAG level and decreases body fat accumulation, compared with TAG oil. To clarify the mechanisms underlying the beneficial effects of DAG, we investigated the metabolic features of DAG in the small intestine with regard to the digestion pathway in the lumen and the TAG-synthesis pathway in the mucosa. When intraduodenally infused as an emulsion, TAG was digested to 1,2-DAG, 2-MAG, and FFA, whereas 1,3-DAG was digested to 1(3)-MAG and FFA. When assessed by the incorporation of [1-¹⁴C]linoleic acid in lipids, the mucosal TAG-synthesis was significantly reduced by DAG infusion compared with TAG infusion. However, the mucosal 1,3-DAG synthesis was remarkably increased in the DAG-infused rats. The total amount of mucosal 1,3-DAG was also increased (4.5-fold) after DAG infusion compared with that after TAG infusion. Next, we examined the synthesis pathway of 1,3-DAG. In cultures of the everted intestinal sacs, 1,3-DAG production required the presence of 1-MAG, suggesting that the 1,3-DAG synthesis was due to acylation of 1(3)-MAG in the DAG-infused rats. Furthermore, measurements of DAG acyltransferase activity indicated that 1,3-DAG was little utilized in TAG synthesis. These findings suggest that features of 1,3-DAG digestion and assimilation in the intestine may be responsible for the reduction of the postprandial serum TAG level by dietary DAG.     

Mevalonate 5-pyrophosphate decarboxylase in isolated villus and crypt cells of chick intestine

Mevalonate 5-pyrophosphate decarboxylase was studied in isolated enterocytes obtained from duodenal, jejunal and ileal villi and crypts. In our assay conditions, decarboxylase activity was linear for 60 min and up to 0.3 mg of protein. The subcellular location of decarboxylase in chick enterocytes was investigated. About 94% of the total activity was recovered in the cytosol. The distribution of enzyme activity in epithelial cells also was studied. Maximal specific activity was found in cell fractions from jejunum followed by ileum and duodenum. About 80% of total activity was recovered in the villus cells, indicating an active role of these cells in cholesterogenesis. Ileal cells showed the highest cholesterol content. In all the intestinal epithelial cells assayed, free cholesterol represented about 95% of the total cholesterol.     

Regulation of cholesterol synthesis in isolated epithelial cells of human small intestine

We have investigated the regulation of cholesterol synthesis in isolated human small intestine epithelial cells (enterocytes). It was established that the amount of cholesterol synthesized increased linearly with the incubation time and the number of cells in the incubation mixture; the synthesis was suppressed by 7-ketocholesterol. Cholic, dehydrocholic, chenodeoxycholic, glycocholic, taurocholic, taurochenodeoxycholic and taurodeoxycholic acids inhibited cholesterol synthesis in enterocytes to different degrees in a dose-dependent manner. Lithocholic acid enhanced the rate of cholesterol synthesis. Deoxycholic acid, methyl ester of cholic acid and cholesterol did not affect the process. No bile acids tested, with the exception of taurodeoxycholic acid, affected fatty acid synthesis in enterocytes. Most bile acids also decreased cholesterol synthesis in cultured human skin fibroblasts. The results obtained make it possible to postulate that cholesterol synthesis in human enterocytes may be subject to a complex regulation by bile acids.     

Fatty acid and sterol synthesis by rat small intestine in vitro

Slices of rat jejunum were incubated with [2-¹⁴C]pyruvate, [1-¹⁴C]acetate, or [³H]H₂O to determine lipogenic activity. Under all conditions studied, pyruvate acted as a better precursor than acetate for fatty acid synthesis but not for the synthesis of sterol. Exogenous glucose significantly (P≤0.05) increased the conversion of both pyruvate and acetate to fatty acids. By contrast, fasting resulted in a decrease (p≤0.05) in lipogenic activity. The highest levels of lipogenesis were observed when [³H]H₂O + glucose at a concentration of 20 mM was used. From such experiments, the absolute rate of fatty acid synthesis in the tissue preparation was calculated: 734±54 nmoles acetyl units incorporated into fatty acids/g tissue/hr.     

Cholesterol metabolism in the liver and intestine of the chick: Effect of dietary cholesterol,taurocholic acid and cholestyramine

The effect of feeding cholesterol, taurocholic acid, or cholestyramine to chicks on cholesterolgenesis from [1-¹⁴C] acetate in liver and intestine was determined in vitro using tissue slices, and in vivo by i.v. injection of [¹⁴C] acetate. The conversion of cholesterol to bile acids in liver in vivo was measured in the same treatments after i.v. injection of [³H] cholesterol. Hepatic cholesterogenesis in vitro and in vivo was depressed by dietary cholesterol and taurocholate and enhanced by cholestyramine. Intestinal cholesterogenesis in vivo was depressed only by taurocholate whereas ileal cholesterogenesis in vitro was reduced by dietary cholesterol. Conversion of cholesterol to bile acids was enhanced by dietary cholesterol and cholestyramine and depressed by taurocholate. Hepatic cholesterol metabolism in the chick appears to be regulated by mechanisms similar to those reported for other species.     

Embryonic vs tumor lipids: II. Changes in phospholipids of developing chick brain,heart, and liver

Brain, heart, and liver tissues were excised from embryos and chicks 10, 13, 16, 19, 22, 27, and 53 days after incubation was initiated and the lipids extracted. The quantitative distribution of the phospholipids and the fatty acid composition of the individual phosphatides were determined for each time period. Each tissue exhibited a distinct phospholipid composition that differed from the composition of egg. Elevated concentrations of particular phosphoglycerides that characterize certain mature tissues were observed at the earliest time period. As development progressed, some phospholipid classes in all tissues showed dramatic change, while others remained relatively constant. Brain showed the most stable composition, while the phosphatides of liver were the most dynamic. Each phospholipid class exhibited a characteristic fatty acid profile that was unique for each tissue. All of the phospholipid classes showed a change in fatty acid composition as development progressed, and, in some tissue, the change was dramatic. The fatty acid composition of brain phosphoglycerides showed the least change, while liver showed the greatest fluctuation. Docosahexaenoic acid and, in most cases, arachidonic acid decreased in the phosphoglycerides with increased development. The decrease in docosahexaenoic acid correlated well with the decreasing mitotic indices of heart and liver cells as development progressed. Comparison of observed abnormal lipid patterns between mature and neoplastic tissue with embryonic tissue lipid profiles suggest that some of the observed abnormalities of neoplasms probably are due to changes in lipid metabolism associated with rapidly proliferating cells, whereas other abnormalities appear to be associated with neoplasia.     

Characterization of calcium-independent cytosolic phospholipase A2 activity in the submucosal regions of rat stomach and small intestine

This study was undertaken to compare the calcium-independent phospholipase A₂ (PLA₂) activities in the cytosols of twelve rat tissues and to determine whether their activities were distinct. 1-O-Alk-1′-enyl-2-[¹⁴C]-oleoyl-sn-glycero-3-phosphocho-line (PlsC) and 1-O-Alk-1′-enyl-2-[¹⁴C]oleoyl-sn-glycero-3-phosphethanolamine (PlsE) were synthesized and used as substrates, instead of phosphatidyl compounds, to exclude hydrolysis by cytosolic PLA₁ activity that could be present in some of the cytosolic preparations. For each tissue, we examined substrate specificity, pH optimum, and effect of adenosine triphosphate (ATP) and ATP analogues. PLA₂ activity was detected in eleven out of the twelve issues examined. Based on substrate specificity and pH optimum, cytosolic calcium-independent PLA₂ were classified in three groups. The first group, which included PLA₂ from small intestine, stomach and spleen, had the highest specific activity with PlsC as substrate (1253, 309 and 75 nmol/mg protein/hour, respectively) and an optimal pH at 6.5. Activity with PlsE as substrate was much lower (20–37%) than with PlsC. The second group of PLA₂ activities included the cytosolic activities from thymus, lung, liver and pancreas that showed lower specific activities for both substrates (14–23 nmol/mg protein/hour with PlsC) and had a broader optimal pH range of 6.1 to 7.5. The cytosols from brain, kidney, heart and muscle comprised the third PLA₂ group that was found to have a higher specific activity with PlsE (5–20 nmol/mg protein/hour) than PlsC and an optimal pH range from 7.4 to 7.9. Since the highest specific activity was found in the cytosol from small intestine, this PLA₂ was examined further. PLA₂ activity was found to be equally distributed in the cytosol of the submucosal portion of duodenum, jejunum and ileum with an optimal pH of 6.1 and a 5-fold higher activity with PlsC than PlsE as substrate. Moreover, this PLA₂ activity was inhibited by treatment with detergents. These results indicate the presence in the submucosal portion of the intestine of a calcium-independent cytosolic PLA₂ with a high specific activity toward PlsC and properties distinct from those described for the PLA₂ found in the intestinal brush-border.     

Lecithin inhibits fatty acid and bile salt absorption from rat small intestine in vivo

During digestion of a fatty meal, long chain free fatty acids (FFA) and lecithin are among the lipids solubilized in intestinal contents as mixed micelles with bile salts. We hypothesized that if lecithin were not hydrolyzed, the mixed micelles would be abnormal, and absorption of FFA and bile salts would be depressed. To test this hypothesis, isolated segments of rat small intestine were infused in vivo with micellar solutions of 2 mMolar linoleic acid and 10 mMolar taurocholate to which was added 3 mMolar 1-palmitoyl, 2-oleoyl lecithin (a common lecithin in bile and food), or 1-palmitoyl lysolecithin (the hydrolytic product of lecithin). Absorption of FFA and bile salt was measured under steady state conditions using a single-pass technique. Lecithin depressed the rate of FFA absorption by 40% (p<0.025) in jejunal and ileal segments whereas lysolecithin was associated with normal rates of FFA absorption. Lecithin also reduced taurocholate absorption from the ileum by 30% (p<0.05). These data support the idea that lecithin may depress FFA and bile salt absorption from the small intestine in pancreatic insufficiency. The following trivial names are used: lecithin (1,2-diacyl-sn-glycero-3-phosphorylcholine); lysolecithin (1-acyl-sn-glycero-3-phosphorylcholine).     

Changes in the chemical composition of surfactant-like particles secreted by rat small intestine in response to different dietary fats

Consumption of dietary oil, viz., corn, fish, coconut, or olive, induced the secretion of surfactant-like particles (SLP) in rat intestine. These lipoprotein particles differ in (i) levels of alkaline phosphatase activity, (ii) lipid composition, and (iii) FA composition in response to feeding of different oils. The secreted particles had similar buoyancy (1.07–1.08 g/mL) and cholesterol/phospholipid molar ratios (0.61–0.72) except that feeding coconut oil to rats produced SLP with a low (0.18) cholesterol/phospholipid molar ratio compared to control animals. It is concluded from these observations that feeding different oils induces the secretion of lipoprotein particles in rat intestine with different chemical compositions.     

金霉素的水解动力学研究

应用紫外分光光度法研究了金霉素在不同温度,不同酸碱条件下及不同水生环境中金霉素的降解情况。结果表明,金霉素较易水解,其水解速率受pH和温度的影响较大。在碱性及中性条件下的水解速率明显大于酸性条件,而高温(70℃)条件下的水解速率远大于室温(20℃);金霉素进入环境水体后,由于各种降解过程综合作用的结果,比用水解试验得出的降解速率要快很多。     

Studies on the role of phospholipids in the triglyceride cycle: II. Turnover of plasma phospholipids in ethionine-treated dogs

The disappearance of native labeled plasma phospholipids (nPLP³²) was abruptly halted in the plasma of normal mongrel dogs shortly after givingdl-ethionine by intraperitoneal injection. In other dogs, chronic feeding of ethionine over a period of three days prior to these measurements prolonged the turnover time of plasma phospholipids. This impairment in the turnover of plasma PL brought about by ethionine administration occurs regardless of sex differences in lipid metabolism or other physiological differences relating to the nutritional status of the animal. The amount of nPLP³² found in the liver at the end of the turnover experiments was also measured. These data suggest that phospholipid transport from plasma to liver is not impaired by ethionine. The possible significance of an ethionine-induced choline deficiency which impairs transport of triglycerides and phospholipids from the liver to plasma is considered in regard to the role of phospholipids in the triglyceride cycle. Presented in part at the 58th Annual AOCS Meeting, New Orleans 1967.     

Metabolism of gossypol in the chick

¹⁴C labeled gossypol was administered to young chickens and the deposition of gossypol plus gossypol decomposition products in the various tissues was evaluated by determination of¹⁴C activity in a scintillation spectrometer. Most of the gossypol activity was recovered in the feces with relatively smaller amounts retained by the tissues. Most of the labeled compound retained was found in the liver, muscle, blood and kidneys, with the highest concentration in the liver. Increasing the protein content of the diet by the addition of fish meal reduced the amount of gossypol plus gossypol decomposition products found in the tissues. Deceased: July 3, 1968.     

The functions of the phospholipids in the animal body

This paper deals with the bearing of some of the recent information on the three proposed functions of the phospholipids: that they are intermediary metabolites in fat metabolism; that they serve as an oxidation-reduction system; and that they are essential elements in cell structure. The rapid turnover of phospholipids in intestinal mucosa, liver and blood plasma indicates an active part in fat metabolism. The slow turnover in other tissues, notably skeletal muscle. points to a non-metabolic function. The amount of phospholipid present in various skeletal muscles is shown to be a function of the relative extent to which they are used. The adaptation seems to be a slow process since forced activity of a muscle produces a comparatively slight increase in phospholipid content.