Dietary Fat Does Not Overcome trans-10, cis-12 Conjugated Linoleic Acid Inhibition of Milk Fat Synthesis in Lactating mice

Trans-10, cis-12 conjugated linoleic acid (CLA) is a potent inhibitor of milk fat synthesis in the cow and similarly reduces milk fat in rodents. The objective of this study was to determine whether dietary fat can overcome CLA inhibition of milk fat concentration in lactating mice. Wild type C57Bl/6J mice (n = 31) were fed semipurified diets containing either low fat (LF; 4% fat) or high fat (HF; 23.6% fat) starting 4–6 days postpartum. Dietary fat was increased by inclusion of high oleic sunflower oil. After 2 days on the experimental diets, lactating dams were orally dosed with either water (control) or trans-10, cis-12 CLA (20 mg/day) for 5 days. CLA treatment decreased pup growth similarly in both HF and LF diets. Milk fat percent was increased over 16% by the HF diet and decreased over 12% by CLA, but there was no interaction of dietary fat and CLA. Both CLA and the HF diet reduced the proportion of short- and medium-chain fatty acids that originate from de novo synthesis, and there was no interaction of diet and CLA. CLA had no effect on the percent of preformed fatty acids, but the HF diet increased their abundance. Dietary fat and CLA both modified mammary expression of lipogenic enzymes and regulators, but no interactions were observed. In conclusion, CLA reduced milk fat concentration and litter growth, but these effects were not overcome by increased dietary fat from high oleic sunflower oil. CLA inhibition of milk fat in the mammary gland is not substrate dependent, and the mechanism is independent from dietary supply of oleic acid.     

trans-10,cis-12 Conjugated Linoleic Acid Improved Growth Performance, Reduced Lipid Deposition and Influenced CPT I Kinetic Constants of Juvenile Synechogobius hasta

trans-10,cis-12 (t10c12) Conjugated linoleic acid (CLA) reduced body lipid deposition in various experimental animals, but the mechanisms involved were still emerging. Carnitine palmitoyltransferase I (CPT I) catalyzes an important regulatory step in lipid metabolism. At present, no studies, to our knowledge, have evaluated the kinetic constants influenced by dietary CLA in fish. In the present study, we tested the hypothesis that changes in body lipid content in fish as a response to dietary t10c12 CLA was related to the change of CPT I kinetic constants [Michaelis constant (K _m), maximal velocity and catalytic efficiency for carnitine and palmitoyl-CoA]. Juvenile Synechogobius hasta were fed three experimental diets with fish oil replaced with 0 (control), 1, or 2 % t10c12 CLA for 8 weeks. Weight gain, specific growth rate and protein efficiency rate increased with dietary t10c12 CLA level. Dietary t10c12 CLA addition significantly reduced lipid contents both in liver and muscle. Dietary CLA addition also improved CPT I activities in muscle but did not significantly influence hepatic CPT I activity. CPT I kinetic parameters (K _m, V _max and catalytic efficiency) were significantly influenced by t10c12 CLA. CPT I catalytic efficiencies with carnitine and palmitoyl-CoA as substrates were higher in muscle and liver of fish fed increasing t10c12 CLA. For the first time, the findings demonstrated effect of dietary CLA addition on CPT I kinetics in fish and supported our starting hypothesis that dietary t10c12 CLA addition induced alterations in CPT I kinetic constants of muscle and liver. Increased CPT I catalytic efficiency might be the main reason for reduced lipid deposition in these tissues by dietary t10c12 CLA supplementation.     

共轭亚油酸的生物合成及相关酶基因的克隆与表达

共轭亚油酸是一类在9-与11-位、10-与12-位或11-与13-位碳原子处有顺式或反式共轭双键的十八碳二烯酸.在共轭亚油酸的各种异构体中,只有c9,t11-18:2和t10,c12-18:2两种异构体具有多种生理活性,如抗癌、抗粥状动脉硬化、抗氧化、降低胆固醇、减肥等.共轭亚油酸异构酶可以专一性地催化亚油酸转化为活性共轭亚油酸异构体,因此近几年来倍受关注,利用生物合成法生产共轭亚油酸已成为该领域的发展趋势.对近年来共轭亚油酸的生物合成以及相关酶基因克隆表达方面的研究进展进行了综述.     

Milk fat synthesis is unaffected by abomasal infusion of the conjugated diene 18:3 isomers cis-6,trans-10, cis-12 and cis-6,trans-8,cis-12

It has been previously established that trans-10, cis-12 CLA is a potent inhibitor of milk fat synthesis. Although the mechanism of this action is not completely understood, it has been speculated that eicosanoid-like metabolites of this isomer formed by the activity of tissue desaturases may be responsible for its activity. The objective of this study was to investigate the effects of an enrichment containing an 18∶3 conjugated diene, produced in the metabolism of trans-10, cis-12 CLA, on milk fat synthesis. Three rumen-fistulated Holstein cows (210±8 d in milk) were randomly assigned in a 3×3 Latin square experiment. Treatments were (i) control, (ii) trans-10, cis-12 CLA supplement (2.1 g/d; positive control), (iii) enrichment providing two conjugated diene 18∶3 isomers (2.6 g/d of cis-6, trans-10, cis-12 and 4.0 g/d of cis-6, trans-8, cis-12) and trans-10, cis-12 CLA (2.1 g/d). Treatments were abomasally infused for 5 d at 4-h intervals, and there was a 7-d interval between periods. Milk yield, dry matter intake, and milk protein yield were unaffected by treatments. In contrast, the trans-10, cis-12 CLA supplement reduced milk fat yield by 27%, whereas the supplement enriched with conjugated diene 18∶3 isomers (treatment iii) had no effect on milk fat yield beyond that attributable to its trans-10, cis-12 CLA content. The transfer efficiency of trans-10, cis-12 CLA into milk fat was 25 and 24% for treatments ii and iii, respectively. At the same time, the abomasally infused conjugated diene 18∶3 isomers were transferred to milk fat with an efficiency of 33 and 41% for cis-6, trans-10, cis-12 and cis-6, trans-8, cis-12 18∶3, respectively. Overall, short-term abomasal infusion of the conjugated diene 18∶3 isomers had no effect on milk fat synthesis, thereby offering no support for an involvement of metabolies of trans-10, cis-12 CLA in the regulation of milk fat synthesis.     

Dietary Conjugated Linoleic Acid-c9t11 Prevents Collagen-Induced Arthritis,Whereas Conjugated Linoleic Acid-t10c12 Increases Arthritic Severity

Two conjugated linoleic acid (CLA) isomers, cis‐9, trans‐11 (CLAc9t11) and trans‐10, cis‐12 (CLAt10c12), reduce inflammation in a number of animal models, including collagen‐induced arthritis (CA). However, little is known about the ability of individual CLA isomers to prevent autoimmune disease onset. Evidence that mixed isomer CLA drives T helper cell (Th) 1 responses suggests that CLA, or a specific isomer, exacerbates onset of Th1 autoimmune diseases. In two experiments, we examined if prior dietary exposure to CLAt10c12 (experiment 1) or CLAc9t11 (experiment 2) affected the incidence or severity of CA. DBA/1 mice were fed a semi purified diet with either 6% corn oil (CO, w/w), 5.75% CO plus 0.25% CLAt10c12, or 5.5% CO plus 0.5% CLAc9t11 prior to arthritis development. Arthritis incidence and severity, anti‐collagen antibodies, paw cytokines, and hepatic fatty acids were measured. CLAt10c12 had no effect on arthritis incidence but increased arthritic severity (42%, P = 0.02); however, CLAc9t11 decreased arthritis incidence 39% compared to CO fed mice (P = 0.01), but had no effect on disease severity. CLAt10c12‐induced increase in anti‐collagen type II IgG antibodies may be a mechanism by which this isomer increased arthritic severity, and CLAc9t11‐induced increase in Th2 paw cytokines (IL‐4 and IL‐10, P ≤ 0.04) may explain how CLAc9t11 reduced the arthritis incidence. While both isomers are well known to reduce inflammation in arthritic mice, these new data suggest isomer differences when fed prior to autoimmune disease.     

A simple method of preparation of methyl trans-10,cis-12- and cis-9,trans-11-octadecadienoates from methyl linoleate

Pure conjugated isomers of linoleic acid were prepared on a large scale by alkali-isomerization of purified methyl linoleate. The methyl esters of alkali-isomerized linoleic acid contained mainly the methyl cis-9,trans-11- and trans-10,cis-12-octadecadienoates (44 and 47%, respectively). These two isomers were then separated and purified by a series of low-temperature crystallizations from acetone. The isomeric purity obtained for the cis-9,trans-11-octadecadienoate isomer was >90% and that of the trans-10,cis-12-octadecadienoate isomer was 89 to 97%. The isolated yield of the two isomers corresponded to 18 and 25.7%, respectively, of the starting material. The structure of the two isomers was confirmed using partial hydrazine reduction, silver nitrate-thin-layer chromatography of the resulting monoenes and gas chromatography coupled with mass spectrometry of the 4,4-dimethyloxazoline derivatives. Fourier transform infrared spectroscopy of the monoenes gave the confirmation of the geometry of each double bond.     

cis-9,trans-11 and trans-10,cis-12 CLA affect lipid metabolism differently in primary white and brown adipocytes of Djungarian hamsters

We explored whether CLA isomers and other C₁₈ FA affect (i) lipid content and FA concentrations in total adipocyte lipids, (ii) FA synthesis from glucose in TAG and phospholipids of primary brown (BAT) and white adipocytes (WAT), and (iii) mRNA expression of uncoupling protein 1 (UCP1) in primary brown adipocytes of Djungarian hamsters (Phodopus sungorus). c9,t11-CLA, oleic, linoleic, and α-linolenic acid increased whereas t10,c12-CLA decreased lipid accumulation in both adipocyte types. t10,c12-CLA treatment affected FA composition mainly in BAT cells. CLA incorporation into lipids, in particular c9,t11-CLA, was higher in BAT. In both cell types, t10,c12-CLA treatment reduced the incorporation of glucose ¹³C carbon into FA of TAG and phospholipids, whereas c9,t11-CLA, linoleic, and α-linolenic acid either did not influence or dose-dependently increased glucose carbon incorporation into FA. UCP1 mRNA expression was inhibited by t10,c12-CLA but increased by c9,t11-CLA, linoleic, and α-linolenic acid. It is concluded that c9,t11-CLA and t10,c12-CLA have distinctly different effects on lipid metabolism in primary adipocytes. The effects of c9,t11-CLA are similar to those of other unsaturated C₁₈ FA. The opposite effects of c9,t11-CLA and t10,c12-CLA are evident in both WAT and BAT cultures; however, brown adipocytes seem to be more susceptible to CLA treatment.     

Effects of cis-9, trans-11 and trans-10, cis-12 CLA isomers on liver and adipose tissue fatty acid profile in hamsters

The aim of the present study was to investigate the effect of cis-9, trans-11 and trans-10, cis-12 CLA on FA composition of TAG in epididymal adipose tissue and liver, and of hepatic phospholipids PL. Twenty-four Syrian Golden hamsters were randomly divided into three groups of eight animals each and fed semipurified atherogenic diets supplemented with either 0.5 g/100g diet of linoleic acid or cis-9, trans-11 or trans-12, cis-9 CLA for 6 wk. Total lipids were extracted, and TAG and PL were separated by TLC. FA profile in lipid species from liver and adipose tissue, as well as in feces, was determined by GC. Trans-10, cis-12 CLA feeding significantly reduced linoleic and linolenic acids in TAG from both tissues, leading to reduced total PUFA content. Moreover, in the epididymal adipose tissue docosenoic and arachidonic acids were significantly increased. In liver PL, although no changes in individual FA were observed, total saturated FA (SFA) were decreased. No changes in TAG and PL FA profiles were induced by the cis-9, trans-11 CLA. TAG and PL incorporated cis-9, trans-11 more readily than trans-11, cis-12 CLA. This difference was not due to differential intestinal absorption, as shown by the analysis of feces. We concluded that only trans-10, cis-12 CLA induces changes in FA composition. Whereas increased PUFA content was observed in either liver or adipose tissue TAG, decreased SFA were found in liver PL. Incorporation of cis-9, trans-11 CLA in TAG is greater than that of trans-10, cis-12 CLA, but this is not due to differences in intestinal absorption.     

Dietary Free Oleic and Linoleic Acid Enhances Neutrophil Function and Modulates the Inflammatory Response in Rats

The high ingestion of oleic (OLA) and linoleic (LNA) acids by Western populations, the presence of inflammatory diseases in these populations, and the importance of neutrophils in the inflammatory process led us to investigate the effects of oral ingestion of unesterified OLA and LNA on rat neutrophil function. Pure OLA and LNA were administered by gavage over 10 days. The doses used (0.11, 0.22 and 0.44 g/kg of body weight) were based on the Western consumption of OLA and LNA. Neither fatty acid affected food, calorie or water intake. The fatty acids were not toxic to neutrophils as evaluated by cytometry using propidium iodide (membrane integrity and DNA fragmentation). Neutrophil migration in response to intraperitoneal injection of glycogen and in the air pouch assay, was elevated after administration of either OLA or LNA. This effect was associated with enhancement of rolling and increased release of the chemokine CINC-2αβ. Both fatty acids elevated l-selectin expression, whereas no effect on β₂-integrin expression was observed, as evaluated by flow cytometry. LNA increased the production of proinflammatory cytokines (IL-1β and CINC-2αβ) by neutrophils after 4 h in culture and both fatty acids decreased the release of the same cytokines after 18 h. In conclusion, OLA and LNA modulate several functions of neutrophils and can influence the inflammatory process.     

Conjugated Linoleic Acid (t-10, c-12) Reduces Fatty Acid Synthesis de Novo, but not Expression of Genes for Lipid Metabolism in Bovine Adipose Tissue ex Vivo

We hypothesized that exogenous fatty acids, and especially or 18:2 trans-10, cis-12 conjugated linoleic acid (CLA), would decrease adipogenic and lipogenic gene expression and de novo fatty acid biosynthesis in intramuscular (i.m.) and subcutaneous (s.c.) adipose tissues. Fresh i.m. and s.c. adipose tissues were collected from the longissimus thoracis muscle of Angus steers at 12, 14, and 16 months of age (n = 4 per time point). Adipose tissue explants were incubated in duplicate for 48 h with 40 μM α-linolenic (ALA), oleic, stearic, trans-vaccenic, or CLA. Adipocyte size, acetate and glucose incorporation into fatty acids in vitro and mRNA levels for C/EBPβ, CPT1β, GPR43, PPARγ, PRKAA1 (AMPKα) and SCD1 were measured following the incubations. PRKAA1 and SCD1gene expression were greater (P < 0.001) in s.c. adipose tissue than in i.m. adipose tissue and acetate incorporation into lipids and C/EBPβ, PPARγ, and SCD1gene expression were greater at 16 months of age than at 12 months of age in i.m. adipose (P < 0.01). C/EBPβ gene expression increased by 16 months of age and PRKAA1 gene expression decreased by 16 months of age in s.c. adipose tissue. All fatty acids increased s.c. adipocyte volumes whereas CLA decreased acetate incorporation into lipids in s.c. adipose tissue (P < 0.05), but none of the fatty acids affected gene expression in i.m. or s.c. adipose tissue (P > 0.10). Thus, CLA depressed de novo fatty acid biosynthesis from acetate but neither CLA nor other fatty acids significantly affected adipogenic or lipogenic gene expression.     

Infrared absorption of methylcis-9,trans-11-, andtrans-10,cis-12-octadecadienoates

The ratio of absorptivity at 10.2 µm and 10.6 µm differs between methylcis-9,trans-11-, andtrans-10,cis-12-octadecadienoates. For thecis-9,trans-11-ester, a_10.2 µm/a_10.6 µm is in the range of 1.1–1.2; for thetrans-10,cis-12-ester, it is 1.3–1.4. These differences in absorptivities are great enough to affect significantly compositions calculated from IR absorption.     

植物乳杆菌共轭亚油酸异构酶基因的克隆、序列分析及重组表达

利用MRS培养基培养植物乳杆菌AS1.555,收集菌体后提取总DNA,用PCR技术扩增其亚油酸异构酶基因,克隆到pQE30质粒载体上,并进行测序.测序结果表明,扩增DNA全长1710 bp,编码569个氨基酸,分子量为64.7 ku.经同源性比较,此核酸序列与罗伊氏乳杆菌、短双歧杆菌、疮疱丙酸杆菌亚油酸异构酶基因核苷酸序列差别较大,推测此基因的来源与上述菌种不同.     

Evidence that the trans-10,cis-12 isomer of conjugated linoleic acid induces body composition changes in mice

We investigated the effects of conjugated linoleic acid (CLA) preparations, which were enriched for the cis-9,trans-11 CLA isomer or the trans-10,cis-12 CLA isomer, on body composition in mice. Body composition changes (reduced body fat, enhanced body water, enhanced body protein, and enhanced body ash) were associated with feeding the trans-10,cis-12 CLA isomer. In cultured 3T3-L1 adipocytes, the trans-10,cis-12 isomer reduced lipoprotein lipase activity, intracellular triacylglycerol and glycerol, and enhanced glycerol release into the medium. By contrast, the cis-9,trans-11 and trans-9,trans-11 CLA isomers did not affect these biochemical activities. We conclude that CLA-associated body composition change results from feeding the trans-10,cis-12 isomer.     

Effects of the individual isomers cis-9,trans-11 vs. trans-10,cis-12 of conjugated linoleic acid (CLA) on inflammation parameters in moderately overweight subjects with LDL-phenotype B

Immune-modulating effects of CLA have been reported in animals, but results are inconsistent. In humans, CLA has shown no effects or only minor effects on immune function. The objective of this study was to evaluate the immune-modulating effects of 3 g cis-9,trans-11 (c9,t11) vs. trans-10,cis-12 (t10,c12) CLA isomers in a population with a high risk of coronary heart disease characterized by moderate overweight (body-mass index, 25–32.5 kg/m²) in combination with LDL-phenotype B (≥35% small LDL cholesterol, density≥1.040 g/mL). After a run-in period of 1 wk, 42 men and women were randomly allocated to the c9,t11 CLA group, the t10,c12 CLA group, or the placebo group. Effects of 13 wk of consumption of 3 g of CLA isomers on cytokine production by ex vivo lipopolysaccharide (LPS)-stimulated peripheral blood mononuclear cells (PBMC) and whole blood, and on plasma C-remononuclear protein (CRP) concentrations were evaluated. To generate hypotheses for future studies, protein expression patterns of 42 cytokines, chemokines, and growth factors were evaluated with an antibody array in pooled, nonstimulated, fasting plasma samples. LPS induced interleukin (IL)-6, IL-8, and tumor necrosis factor-α production by PBMC, and whole blood as well as plasma CRP concentrations were not significantly changed by the c9,t11, and the t10,c12 CLA isomers. The cytokine expression profile in nonstimulated plasma suggested that both CLA isomers induced a specific inflammatory signature, in which the c9,t11 CLA group showed more activity in terms of numbers of proteins regulated. We conclude that daily consumption of 3 g of c9,t11 or t10,c12 CLA isomer did not affect LPS-stimulated cytokine production by PBMC or whole blood and plasma CRP levels. Inflammatory signatures in fasting, nonstimulated plasma as determined by an antibody array may indicate enhanced immune function by both CLA isomers.     

Evidence that commercial calf and horse sera can contain substantial amounts of trans-10,cis-12 conjugated linoleic acid

We analyzed fetal calf, newborn calf, horse, and adult cow sera for conjugated linoleic acid (CLA). All sera samples contained CLA, but the amounts varied. The predominant isomer was cis-9,trans-11 CLA but some samples appeared to contain substantial amounts of an isomer with the retention time of trans-10,cis-12 CLA.     

Improved Estimation of Oil,Linoleic and Oleic Acid and Seed Hull Fractions in Safflower by NIRS

Near-infrared reflectance spectroscopy (NIRS) of intact seeds allows the non-destructive estimation of seed quality parameters which is highly desirable in plant breeding. Together with yield, oil content and quality, a main aim in safflower (Carthamus tinctorius L.) breeding is the selection of genotypes with a low percentage of empty seeds even under cooler climates. We developed NIRS calibrations for seed oil content, oleic and linoleic acid content, the seed hull fraction and the percentage of empty seeds using seed meal and intact seeds. For the different calibrations 108–534 samples from a safflower breeding program with lines adapted to German conditions, were analyzed with reference analyses (Soxhlet, gas chromatography), and scanned by NIRS as intact seeds and seed meal. Calibration equations were developed and tested through cross validation. The coefficient of determination of the calibration (R ²) for intact seeds ranged from 0.91(oil content), 0.90 (seed hull fraction), 0.84 (empty seeds), 0.73 (linoleic acid) to 0.68 (oleic acid). The coefficient of determination of the cross validation was higher for seed meal than for intact seeds except for the parameter seed hull fraction. The results show that NIRS calibrations are applicable in safflower breeding programs for a fast screening.     

Desaturation indices in liver, muscle, and bone of growing male and female mice fed trans-10, cis-12 conjugated linoleic acid

trans-10, cis-12-CLA (t10,c12-CLA) inhibits lipid deposition in adipose tissue of many species, but it also enhances lipid deposition in liver. We evaluated effects of dietary t10,c12-CLA content and gender on carcass composition, FA profile of selected tissues, and expression of FA synthase (FAS) and stearoyl-CoA desaturase-1 (SCD) mRNA in adipose tissue. Male and female (63 of each) CD-1 mice were assigned a diet containing 0.0, 0.15, or 0.30% t10,c12-CLA at 4 wk of age. Seven mice per dietary group within gender were sacrificed after 2,4 or 6 wk. The CLA isomer caused dose-dependent reductions in dry carcass weight and fat content, without altering protein content, but carcass fat and epididymal fat pad weights of males were reduced to a greater extent than carcass fat and inguinal fat pad weights of females. FAS and SCD mRNA in adipose tissue was more abundant in females than males, but expression in both genders decreased as the t10, c12-CLA content of the diet increased. Although the weight of gastrocnemius muscle was not influenced by diet, total FA content of the muscle of both genders decreased in response to dietary t10,c12-CLA content. Femur weight of male mice increased as the t10,c12-CLA content of the diet increased, but the weight increase was associated with a reduction in total FA content. The Δ9 desaturation indices for muscle and femur suggested a linear reduction in SCD activity, whereas Δ9 indices for liver indicated linear enhancement of SCD activity. Overall, results suggested that growing male mice were more susceptible than females to t10,c12-CLA inhibition of lipid deposition.