10-12-octadecadienoic-acid and 9-11-linoleic-acid

Conjugated linoleic acid (CLA) has been shown to inhibit carcinogenesis and atherosclerosis, enhance immunologic function while protecting against the catabolic effects of immune stimulation, affect body composition change (reducing body fat gain while enhancing lean body mass gain), and stimulate the growth of young rats. We discuss possible biochemical mechanisms that underlie these physiological effects. We emphasize the importance of considering the effects, both individually and combined, of the two CLA isomers (cis-9, trans-11 CLA and trans-10, cis-12 CLA) that have been shown to exhibit biological activity and which appear to exert their effects via different biochemical mechanisms.

Topics: Animals; Anticarcinogenic Agents; Arteriosclerosis; Body Composition; Drug Design; Linoleic Acids; Linoleic Acids, Conjugated; Molecular Conformation; Rats

Conjugated dienoic linoleate (CLA), a linoleic acid derivative, has received considerable attention as a chemoprotective agent in the past few years because it has been shown experimentally to inhibit rat mammary tumorigenesis, mouse forestomach neoplasia, and mouse skin carcinogenesis. CLA has several unique structural and functional properties resulting in chemical and physiological effects that are different from those of all-cis, nonconjugated polyunsaturated fatty acids. In turn, these unique qualities appear to modulate cellular processes involved in carcinogenesis. This review will introduce the chemical background of conjugated dienoic linoleate, examine findings describing its chemoprotective qualities, present possible mechanisms of chemoprotection, and correlate the possible significance of dietary CLA modulation to carcinogenesis to humans.

Topics: Animals; Anticarcinogenic Agents; Humans; Linoleic Acids; Linoleic Acids, Conjugated; Neoplasms, Experimental; Nutritional Physiological Phenomena

Conjugated linoleic acid (CLA) refers to geometric and positional isomers of linoleic acid. Animal studies have shown that CLA modulates the immune system and suggest that it may have a therapeutic role in inflammatory disorders. This double-blind placebo-controlled intervention trial investigated the effects of CLA supplementation on indices of immunity relating to cardiovascular disease (CVD) in a cohort of healthy middle-aged male volunteers. Subjects were randomly assigned to supplement their diet with 2.2 g 50:50 isomeric blend of cis 9, trans 11 (c9, t11)-CLA and trans 10, cis 12 (t10, c12)-CLA or placebo daily for 8 weeks. Interleukin (IL) 2, IL-10 and tumour necrosis factor (TNF) alpha were measured in the supernatant of cultured unstimulated and concanavalin A (Con A)-stimulated peripheral blood mononuclear cells (PBMC) by ELISA. Serum IL-6 and plasma CRP were measured by ELISA and plasma fibrinogen by automated clotting assay. Gene expression was investigated by real-time RT-PCR. CLA supplementation significantly reduced Con A-stimulated PBMC IL-2 secretion (37.1%; P=.02). CLA supplementation had no significant effect on transcription of IL-2. CLA supplementation had no direct significant effects on PBMC TNFalpha or IL-10 secretion. Other inflammatory markers associated with CVD, including IL-6, CRP and fibrinogen, were not affected by CLA supplementation. This study showed that CLA supplementation reduced PBMC IL-2 secretion from Con A-stimulated PBMC but lacked effect on other markers of the human inflammatory response.

Topics: Adult; Concanavalin A; Double-Blind Method; Humans; Interleukin-10; Interleukin-2; Leukocytes, Mononuclear; Linoleic Acids, Conjugated; Male; Middle Aged; Tumor Necrosis Factor-alpha

Obesity-related hypertension may be caused by activation of the local adipose tissue renin-angiotensin system, resulting in exaggerated production of the vasoconstrictor angiotensin II. Additionally, secretion of adiponectin from adipose tissue, which prevents endothelial dysfunction, is altered in obesity. Consumption of conjugated linoleic acid (CLA) has been shown to modulate cytokine release from adipocytes and positively influence blood pressure in younger rats, but its physiological actions in older models with established hypertension and isomer-specific effects on adipocyte size remain to be determined. Therefore, we investigated the effects of CLA isomers on adipocyte size in relation to blood pressure and adipokine production by hypertrophic adipocytes in older fa/fa Zucker rats with established hypertension. fa/fa Zucker rats were fed with cis(c)9, trans(t)11-CLA or t10, c12-CLA isomers for 8 weeks and compared with lean and obese rats fed with the control diet. Blood pressure and adipocyte size were subsequently measured. Collagenase-isolated adipocytes were size-separated and angiotensinogen and adiponectin protein levels quantified by Western blotting. The t10, c12-CLA group had reduced blood pressure, fewer large adipocytes and increased serum adiponectin. Angiotensinogen was present at higher levels in the large adipocytes, whereas the converse was observed for adiponectin. The beneficial effects of the t10, c12-CLA isomer on blood pressure and adipocyte size in vivo may be due to its ability to reduce the number of large adipocytes, which alters the levels of vasoactive molecules secreted from adipose tissue.

Topics: Adipocytes; Adiponectin; Adipose Tissue; Angiotensinogen; Animals; Anti-Obesity Agents; Antihypertensive Agents; Blood Pressure; Blotting, Western; Collagenases; Dietary Fats; Hypertension; Hypertrophy; Isomerism; Linoleic Acids, Conjugated; Male; Obesity; Rats; Rats, Zucker

Previous studies have demonstrated that the intake of a 1% conjugated linoleic acid (CLA) diet in an 80:20 mixture of cis-9,trans-11 and trans-10,cis-12 exerts age-specific effects on the immune system: immunoglobulin enhancement and proliferative down-modulation in neonatal and adult rats, respectively. The present study evaluates the influence of the same diet on antibody synthesis of early infant Wistar rats during suckling and/or after weaning. Dietary supplementation was performed during suckling and early infancy (4 weeks), only during suckling (3 weeks), or only in early infancy (1 week). CLA content in plasma and serum immunoglobulin (Ig) G, IgM and IgA concentration were determined. Proliferation, cytokines and Ig production were evaluated on isolated splenocytes. Cis-9,trans-11- and trans-10,cis-12-CLA isomers were detected in the plasma of all CLA-supplemented animals, and the highest content was quantified in those rats supplemented over the longest period. These rats also exhibited higher concentrations of serum IgG, IgM and IgA. Moreover, splenocytes from CLA-supplemented rats showed the highest IgM and IgG synthesis and interleukin (IL)-6 production, whereas their proliferative ability was lower. In summary, in infant rats, we observed both the enhance antibody synthesis previously reported in neonates, and the reduced lymphoproliferation previously reported in adults.

Topics: Animal Nutritional Physiological Phenomena; Animals; Animals, Newborn; Cell Proliferation; Cytokines; Diet; Dietary Supplements; Female; Immune System; Immunoglobulin A; Immunoglobulin G; Immunoglobulin M; Linoleic Acids, Conjugated; Lymphocytes; Pregnancy; Rats; Rats, Wistar; Spleen

Conjugated linoleic acid (CLA) is a polyunsaturated fatty acid that has numerous biologic activities. Previous studies in rodents demonstrated that chronic intake of CLA t10,c12 or CLA c9,t11 isomers perturbs the metabolism of retinoids (vitamin A and its derivatives). Specifically, although both isomers increased liver retinoid levels, only CLA t10,c12 also stimulated hepatic retinol secretion into the bloodstream. Given that retinoid homeostasis in mammalian serum and tissues is crucial to maintain health, it is important to gain more insights into the mode of action of this nutrient-nutrient interaction. Here we hypothesized that an acute administration of either CLA isomer may also influence vitamin A metabolism. By gavaging wild-type and retinol-binding protein knockout mice with an oral bolus of radiolabeled retinol containing 1 of these 2 isomers, we showed that both CLA t10,c12 and CLA c9,t11 rapidly enhance hepatic uptake of dietary vitamin A and its resecretion from the liver in the form of retinol bound to retinol-binding protein. Indeed, in mice lacking this protein, the sole specific carrier for retinol in the circulation, this latter effect was blunted. In addition, by using a pharmacologic inhibitor of the clearance of chylomicrons, which distribute recently ingested vitamin A and lipids throughout the body, we provided evidence that CLA intake might rapidly enhance intestinal absorption of dietary vitamin A. These data demonstrate the existence of multiple levels of interaction between dietary CLA and retinoid metabolism and warrant further studies to understand the molecular mechanisms underlying these effects and their implications for human health.

Topics: Animals; Intestinal Absorption; Linoleic Acids, Conjugated; Liver; Male; Mice; Mice, Inbred C57BL; Mice, Knockout; Retinoids; Retinol-Binding Proteins; Tissue Distribution; Vitamin A

Conjugated linoleic acid (CLA) describes a group of isomers of linoleic acid and has variable effects on bone formation and adiposity in vivo and in vitro. The variability may be due to individual effects of the predominant bioactive 9cis,11trans (9,11) and 10trans,12cis (10,12) CLA isomers. Osteoblasts and adipocytes are derived from mesenchymal stem cells (MSCs), and bone loss is accompanied by an increase in marrow adiposity. Osteoblast differentiation from MSCs requires activation of Wnt/beta-catenin signaling by Wnt10b, which inhibits adipocyte differentiation by suppressing CCAAT/enhancer-binding protein (C/EBP) alpha. The objective of this study was to determine if 9,11 and 10,12 CLA affect osteoblast and adipocyte differentiation from MSCs and to determine whether any effects are associated with changes in Wnt10b and C/EBPalpha expression. Osteoblast differentiation was assessed by calcium deposition, alkaline phosphatase (ALP) activity, and the expression of Wnt10b, runx2 and osteocalcin. Adipocyte differentiation was assessed by oil red O staining and C/EBPalpha, PPARgamma and FABP4 expression. Compared to vehicle, 9,11 CLA decreased calcium deposition ( approximately 15%), increased oil red O staining ( approximately 21-28%) and increased FABP4 (AP2) expression ( approximately 58-75%). In contrast, 10,12 CLA increased calcium deposition ( approximately 12-60%), ALP activity ( approximately 2.1-fold) and the expression of Wnt10b ( approximately 60-80%) and osteocalcin ( approximately 90%), but decreased oil red O staining ( approximately 30%) and the expression of C/EBPalpha ( approximately 24-38%) and PPARgamma ( approximately 60%) (P<.05). Thus, our findings demonstrate isomer-specific effects of CLA on MSC differentiation, and suggest that 10,12 CLA may be a useful therapeutic agent to promote osteoblast differentiation from MSCs.

Topics: Adipocytes; Cell Differentiation; Female; Humans; Linoleic Acids, Conjugated; Mesenchymal Stem Cells; Osteoblasts; Stereoisomerism; Young Adult

In vitro work suggests that conjugated linoleic acid (CLA) isomers (c9,t11 and t10,c12) are cytotoxic to human breast cancer cells, however the mechanism remains unknown. Using human MCF-7 breast cancer cells, we examined the effects of c9,t11 and t10,c12 CLA compared to oleic acid (OA), linoleic acid (LA), or untreated cells on cell membrane phospholipid composition, cell survival, and the insulin-like growth factor-I (IGF-I) and the downstream insulin receptor substrate-1 (IRS-1). Both CLA isomers were incorporated into membrane phospholipids (p < 0.05). Compared to untreated cells, c9,t11 or t10,c12 CLA significantly reduced the metabolic activity of IGF-I stimulated MCF-7 cells, increased lactate dehydrogenase (LDH) release, and decreased cellular concentrations of the IGF-I receptor (IGF-IR) and insulin receptor substrate-1 (p < 0.05). Incubation with t10,c12 CLA also reduced the levels of phosphorylated IGF-1R. The effects on all of these measures were greater (p < 0.05) for t10,c12 CLA compared to c9,t11 CLA. There were few differences between LA-treated and c9,t11 CLA-treated cells, whereas cellular metabolic activity, LDH release, and IGF-IR concentrations differed between t10,c12 CLA-treated and LA-treated cells (p < 0.05). OA stimulated growth compared to the untreated condition (p < 0.05). In summary, this study demonstrated that the t10,c12 CLA isomer inhibits growth of MCF-7 cells and suggested that this may be mediated through incorporation into cellular phospholipids and interference with the function of IGF-I and related signaling proteins.

Topics: Breast Neoplasms; Cell Line, Tumor; Cell Membrane; Cell Proliferation; Female; Humans; Insulin Receptor Substrate Proteins; Insulin-Like Growth Factor I; Linoleic Acids, Conjugated; Phospholipids; Receptor, IGF Type 1

Mixed CLA isomers variably affect bone resorption in animals and decrease osteoclast formation and activity in murine osteoclasts. These variable effects may be due to the different isomers present in commercial preparations of CLA, and the effects of the predominant individual isomers, 9cis,11trans (9,11) and 10trans,12cis (10,12) CLA are not clear. The objectives of this study were to determine the effects of the individual CLA isomers on osteoclast formation and activity from human CD14+ monocytes, and to determine whether any changes are accompanied by changes in cathepsin K, matrix metalloproteinase-9 (MMP-9), receptor activator of NF-kappaB (RANK) and tumour necrosis factor alpha (TNFalpha) gene expression. Osteoclasts were identified as TRAP+ multinucleated cells. Osteoclast activity was quantified by the amount of TRAP in the cultured media.. At 50 microM, 9,11 CLA inhibited osteoclast formation by approximately 70%, and both 9,11 and 10,12 CLA decreased osteoclast activity by approximately 85-90%. Both isomers inhibited cathepsin K (50 microM 9,11 by approximately 60%; 10,12 by approximately 50%) and RANK (50 microM 9,11 by approximately 85%; 50 microM 10,12 by approximately 65%) expression, but had no effect on MMP-9 or TNFalpha expression.. 9,11 CLA inhibits osteoclast formation and activity from human cells, suggesting that this isomer may prevent bone resorption in humans. Although 10,12 CLA did not significantly reduce osteoclast formation, it reduced osteoclast activity and cathepsin K and RANK expression, suggesting that this isomer may also affect bone resorption.

Topics: Bone Resorption; Cathepsin K; Cathepsins; Gene Expression Regulation; Humans; Linoleic Acids, Conjugated; Lipopolysaccharide Receptors; Matrix Metalloproteinase 9; Monocytes; Osteoclasts; Receptor Activator of Nuclear Factor-kappa B; Stereoisomerism; Tumor Necrosis Factor-alpha

Conjugated linoleic acid (CLA) is a collective term describing a mixture of positional and geometric isomers of linoleic acid with two conjugated double bonds. Among these compounds two isomers cis9, trans11 and trans10, cis12 have received considerable attention. They are present in milk and meat fat of ruminants and the cis9, trans11 (rumenic acid) is the most abundant with lesser amounts of the trans10, cis12 isomer. A considerable number of papers suggest anticarcinogenic properties of CLA, including their ability to suppress the growth of different cancer cell lines in vitro. It was also found that these isomers may act through different mechanisms to inhibit carcinogenesis. In view of the above, the objective of this paper was evaluation of isomer-specific effects of the natural CLA isomers i.e. cis9, trans11 and trans10, cis12 on proliferation of mammary cancer cell lines. Two mammary cancer cell lines were used: MCF-7 and T47D (ATCC Collection). The cells were incubated with both CLA isomers: cis9,trans11 or trans10, cis12 within the range of 5-200 microM for 24-120 h. There were no toxic effects of any of the isomers in the range of 5-100 microM, as indicated by the Cytotoxicity Detection Kit (Roche) and cells proliferation was determined in these experimental conditions. Proliferation of cells was determined using Cell Proliferation ELISA BrdU (5' bromo-2' deoxyuridine), based on incorporation of BrdU to DNA of growing cells. The results obtained indicate that both isomers suppress the proliferation of the studied mammary cancer cell lines i.e. MCF-7 and T47D, especially when treated with the CLA isomers for 48 h or longer. Of the studied lines the strongest growth-suppressing function approximately 65% was observed for the line T47D.

Topics: Apoptosis; Breast Neoplasms; Cell Division; Cell Line; Cell Line, Tumor; Cell Proliferation; Female; Humans; Linoleic Acids, Conjugated

The objective of this study was to evaluate potential anti-atherogenic properties of hen eggs enriched naturally with conjugated linoleic acid (CLA) isomers (cis-9, trans-11 and trans-10, cis-12). Eighteen apoE and LDL receptor double-knockout mice (apoE/LDLR- / - ), at the age of 4 months with pre-established atherosclerosis, were randomly assigned to three experimental groups (n 6) and fed AIN-93G-based diets for the next 2 months. The experimental diets were: AIN-93G+ CLA-free egg-yolk powder (control); AIN-93G+ CLA-free egg-yolk powder +0.1 % CLA (CLA-supplemented eggs); and AIN-93G+ CLA-enriched egg-yolk powder, providing 0.1 % CLA (CLA-enriched eggs). For assessment of anti-atherogenic properties of CLA-enriched or CLA-supplemented eggs the following criteria were used: (1) serum lipid profile; (2) development of atherosclerosis; and (3) composition of atherosclerotic plaque. CLA-enriched eggs, compared with CLA-supplemented eggs, reduced significantly (P < 0.05) total plasma cholesterol in the mice. At the same time, both CLA-supplemented eggs and CLA-enriched eggs tended to decrease the size of atherosclerotic plaque in aortic roots of mice. Most importantly, atherosclerotic plaques of mice fed CLA-enriched eggs showed significantly (P < 0.05) reduced number of atherogenic macrophages and increased area occupied by smooth muscle cells in atherosclerotic lesions. In conclusion, CLA-enriched eggs exerted an anti-inflammatory effect more effectively than CLA-supplemented eggs. This anti-inflammatory effect can be considered their major functional claim that warrants further exploitation.

Topics: Animals; Aorta; Aorta, Thoracic; Apolipoproteins E; Atherosclerosis; Chickens; Diet; Dietary Supplements; Eggs; Immunohistochemistry; Linoleic Acids, Conjugated; Lipids; Mice; Mice, Knockout; Random Allocation; Receptors, LDL

We have previously shown that the 9c,11t-conjugated linoleic acid (CLA) concentration was always significantly higher than the 10t,12c-CLA concentration following the administration of these compounds to mice and rats, and considered that structural differences between the conjugated double bonds in these isomers affected absorption in the small intestine. This study investigates the absorption of CLA in the rat intestine by a lipid absorption assay of lymph from the thoracic duct. In Study 1, we used safflower oil and a triacylglycerol form of CLA (CLA-TG), while in Study 2, we used 9c,11t-CLA and 10t,12c-CLA. The cumulative recovery of CLA was lower than that of linoleic acid until two hours after sample administration. There was no difference in the extent of lymphatic recovery of 9c,11t-CLA and 10t,12c-CLA after the administration of CLA-TG, 9c,11t-CLA, and 10t,12c-CLA to the rats, suggesting that geometrical and positional isomerism of the conjugated double bonds did not influence the absorption.

Topics: Animals; Intestinal Absorption; Kinetics; Linoleic Acids, Conjugated; Lymph; Rats; Thoracic Duct

This study investigated the incorporation of cis-9,trans-11 conjugated linoleic acid (c9,t11 CLA) and trans-10,cis-12-CLA (t10,c12 CLA) into plasma and peripheral blood mononuclear cell (PBMC) lipids when consumed as supplements highly enriched in these isomers. Healthy men (n = 49, age 31 +/- 8 years) consumed one, two, and four capsules containing approximately 600 mg of either c9,t11 CLA or t10,c12 CLA per capsule for sequential 8 week periods followed by a 6 week washout before consuming the alternative isomer. Both isomers were incorporated in a dose-dependent manner into plasma phosphatidylcholine (PC) (c9,t11 CLA r = 0.779, t10,c12 CLA r = 0.738; P < 0.0001) and cholesteryl ester (CE) (c9,t11 CLA r = 0.706, t10,c12 CLA r = 0.788; P < 0.0001). Only t10,c12 CLA was enriched in plasma nonesterified fatty acids. Both c9,t11 CLA and t10,c12 CLA were incorporated linearly into PBMC total lipids (r = 0.285 and r = 0.273, respectively; P < 0.0005). The highest concentrations of c9,t11 CLA and t10,c12 CLA in PBMC lipids were 3- to 4-fold lower than those in plasma PC and CE. These data suggest that the level of intake is a major determinant of plasma and PBMC CLA content, although PBMCs appear to incorporate both CLA isomers less readily.

Topics: Adult; Cholesterol Esters; Cross-Over Studies; Dietary Supplements; Double-Blind Method; Fatty Acids; Humans; Leukocytes, Mononuclear; Linoleic Acids; Linoleic Acids, Conjugated; Lipids; Male; Phosphatidylcholines

The effects of four conjugated linoleic acid (CLA) isomers on in vitro collagen-induced human platelet aggregation and thromboxane (TXB(2), the inactive metabolite of the proaggregatory TXA(2)) production were examined. As the free fatty acid (FFA), 9t, 11t-CLA was the most effective inhibitor of these two processes (I(50)s of 2.2 and 4 microM, respectively) and the 9c, 11c-CLA was the least effective (I(50)s of 8.3 and 37 microM) of the isomers tested. When platelets were preesterified with either 25 microM 9t, 11t-CLA or 9c, 11c-CLA, CLA incorporation in total platelet lipids increased from 0.24% to 0.31% and 0.38%, and most of this increase was found to be in the phosphatidyl choline and phosphatidyl ethanolamine subclasses. The decrease in arachidonic acid (AA) content in total fatty acids or phospholipids was an order of magnitude greater. Furthermore, no significant differences between platelets prelabeled with either 9t, 11t- or 9c, 11c-CLA in the inhibition of collagen-induced aggregation and TXB(2) formation were observed. However, platelets prelabeled with 9c, 11c-CLA stimulated basal TXB(2) production (4-fold) which was not observed with platelets pretreated with either 9t, 11t-CLA, linoleic acid or stearic acid. This enhancement was associated with a 2.4-5-fold increase in the release of endogenous AA. Our results suggest that the presence of a conjugated cis, cis double bond appears to change the lipid environment sufficiently to stimulate the basal platelet phospholipase activity, which in turn increases the formation of TXB(2).

Topics: Arachidonic Acid; Blood Platelets; Collagen; Enzyme Activation; Humans; Isomerism; Linoleic Acids; Linoleic Acids, Conjugated; Phospholipases; Platelet Aggregation; Platelet Aggregation Inhibitors; Thromboxane B2

There is increasing evidence that individual isomers of conjugated linoleic acid (CLA) may have unique biological or biochemical effects. A primary objective of this study was to determine whether there might be differences in the anticancer activity of 9,11-CLA and 10,12-CLA. This was achieved by evaluating the reduction in premalignant lesions and carcinomas in the mammary gland of rats that had been treated with a single dose of methylnitrosourea and given 0.5% of either highly purified CLA isomer in the diet. Our results showed that the anticancer efficacies of the two isomers were very similar. At 6 wk after carcinogen administration, the total number of premalignant lesions was reduced by 33-36%. At 24 wk, the total number of mammary carcinomas was reduced by 35-40%. The concentration of each CLA isomer and its respective metabolites was analyzed in the mammary fat pad. Tissue level of 10,12-CLA was much lower than that of 9,11-CLA. The pool of metabolites from each isomer was very similar between the two groups and represented only a small fraction of total conjugated diene fatty acids. Feeding of 9,11-CLA resulted in minimal changes in other unsaturated fatty acids. In contrast, feeding of 10,12-CLA produced a wider spectrum of perturbations. Small but significant increases in 16:1 and 16:2 were detected; these were accompanied by decreases in 20:2, 20:3, 20:4, 22:4, and 22:6. The above observation suggests that 10,12-CLA might be more potent than 9,11-CLA in interfering with elongation and desaturation of linoleic and linolenic acids. In summary, our study showed that, at the 0.5% dose level, the anticancer activity of 9,11-CLA and 10,12-CLA was very similar, even though accumulation of 10,12-CLA in the mammary tissue was considerably less than that of 9,11-CLA. These confounding changes of the other unsaturated fatty acids in contributing to the effect of 10,12-CLA need to be clarified.

Topics: Animals; Carcinoma, Intraductal, Noninfiltrating; Fatty Acids, Unsaturated; Female; Linoleic Acids; Linoleic Acids, Conjugated; Mammary Glands, Animal; Mammary Neoplasms, Experimental; Methylnitrosourea; Precancerous Conditions; Rats; Rats, Sprague-Dawley; Time Factors

Conjugated linoleic acid (CLA) isomers are potent inhibitors of mammary tumor cell growth. Evidence suggests that CLA modulates essential fatty acid (EFA) metabolism; however, it is not clear which parts of this pathway are important regulatory points modulated by CLA. Enriched mixtures of D9-cis,11-trans (D9c,11t)- and D10-trans,12-cis (D10t,12c)-18:2 were used to assess outcome measures of EFA metabolism pertaining to membrane phospholipid incorporation, tumor cell growth, and prostaglandin E2 (PGE2) synthesis in the MDA-MB-231 mammary tumor cell line. Tumor cells were treated with linoleic acid (LA), an equal mixture (Mix), or enriched preparations of D9c,11t- or D10t,12c-18:2. Treatment with Mix or the enriched mixture of D10t,12c-18:2 significantly inhibited the synthesis of arachidonic acid (AA) from LA, resulting in increased levels of LA and decreased levels of AA in membrane phosphatidylcholine and phosphatidylethanolamine (P < 0.05). LA and AA levels were not altered in cells treated with enriched D9c,11t-18:2 and were similar to those in LA control treated cells. All CLA treatments reduced [3H]thymidine uptake, an indicator of tumor cell growth, by more than one-half relative to LA controls. MDA-MB-231 cells challenged with AA in the presence of all CLA mixtures resulted in significantly reduced PGE2 synthesis relative to controls treated with LA (P < 0.05). It is evident that individual isomers exert inhibitory effects at specific steps of EFA metabolism, which correspondingly leads to a reduction in PGE2 synthesis and, ultimately, tumor growth.

Topics: Analysis of Variance; Arachidonic Acid; Breast Neoplasms; Dinoprostone; Female; Glycerophospholipids; Humans; Linoleic Acid; Linoleic Acids; Linoleic Acids, Conjugated; Stereoisomerism; Time Factors; Tumor Cells, Cultured

The objective of this study was to determine the effects of cis-9,trans-11 and trans-10,cis-12 CLA on the fatty acid desaturation in a human hepatoma cell line, HepG2. Therefore, experiments were conducted in which HepG2 cells were incubated with various concentrations of those fatty acids and the concentrations of fatty acids in various lipid fractions of HepG2 cells were determined. In the presence of linoleic acid as substrate, cells treated with 25 micromol/L of trans-10,cis-12 CLA had lower ratios of dihomo-gamma-linoleic acid to linoleic acid and of arachidonic acid to linoleic acid in phospholipids than control cells; with alpha-linolenic acid as substrate, they had a lower ratio of eicosapentaenoic acid to alpha-linolenic acid in phospholipids than control cells. Cells treated with cis-9,trans-11 CLA did not differ in these ratios from control cells. Cells treated with trans-10,cis-12 CLA had also a markedly lower ratio of monounsaturated fatty acids (MUFA) to saturated fatty acids (SFA) in lipids than control cells; cells treated with cis-9,trans-11 CLA had a slightly lower MUFA:SFA ratio than control cells. These findings suggest that trans-10,cis-12 CLA suppresses Delta9-, Delta6- and Delta5-desaturation in HepG2 cells; cis-9,trans-11 CLA slightly reduces Delta9-desaturation but does not inhibit Delta6- and Delta5-desaturation. Moreover, HepG2 cells treated with 100 micromol/L of trans-10,cis-12 CLA released larger amounts of 6-keto-prostaglandin F(1alpha) and prostaglandin F(2alpha) than control cells. Treatment of cells with cis-9,trans-11 CLA did not alter the release of these eicosanoids compared with control cells. In conclusion, this study suggests that trans-10,cis-12 CLA has significant effects on the metabolism of essential fatty acids in HepG2 cells, whereas cis-9, trans-11 CLA does not have any effect in this respect.

Topics: alpha-Linolenic Acid; Eicosanoids; Fatty Acids; Hepatocytes; Humans; Isomerism; Linoleic Acids; Linoleic Acids, Conjugated; Lipid Metabolism; Tumor Cells, Cultured

Conjugated linoleic acid (CLA) is a collective term for a group of positional and geometric conjugated dienoic isomers of linoleic acid. CLA has been shown to have strong inhibitory effects on mammary carcinogenesis both in vitro and in vivo. In this study, we investigated the regulation of human stearoyl-CoA desaturase (SCD, EC 1.14.99.5) expression by CLA in human breast cancer cell lines, MDA-MB-231 and MCF-7. Treatment of the cells with the cis-9,trans-11 and trans-10,cis-12 CLA isomers (45 microM) did not repress SCD mRNA in both MDA-MB-231 and MCF-7 cells. However, the cis-9,trans-11 and trans-10,cis-12 CLA isomers significantly decreased SCD protein levels and SCD activity in MDA-MB-231 cells. In MCF-7 cells, both isomers did not affect protein levels, but they inhibited SCD activity. These results suggest that in MDA-MB-231 cells the cis-9,trans-11 and trans-10,cis-12 CLA isomers regulate human SCD by reducing SCD protein levels, while in MCF-7 cells both isomers have a direct inhibitory effect on SCD enzyme activity.

Topics: Anticarcinogenic Agents; Blotting, Northern; Blotting, Western; Breast Neoplasms; Fatty Acids; Humans; Linoleic Acids; Linoleic Acids, Conjugated; Lipid Metabolism; Microsomes; RNA; RNA, Messenger; Stearoyl-CoA Desaturase; Stereoisomerism; Tumor Cells, Cultured

Stearoyl-CoA desaturase (SCD) catalyzes the rate-limiting step in the cellular synthesis of monounsaturated fatty acids mainly oleate (C18:1) and palmitoleate (C16:1) which are the major monounsaturated fatty acids of membrane phospholipids, cholesterol esters, waxes, and triglycerides. Several SCD isoforms exist in the mouse whereas the human has one well-characterized SCD gene. The trans-10,cis-12 isomer of conjugated linoleic acid has been previously shown to repress the expression of the mouse SCD1 gene isomer by decreasing SCD gene expression as well as by direct inhibition of SCD enzyme activity. We studied the regulation of human stearoyl-CoA desaturase (SCD) expression by conjugated linoleic acid (CLA) in cultured human hepatoblastoma cell line, HepG2. Treatment of the cells with the trans-10,cis-12 CLA isomer did not cause changes in the SCD gene transcription, mRNA and protein levels. However, this isomer decreased both the SCD activity as well as the levels of monounsaturated fatty acids. The other major CLA isomer, cis-9,trans-11 CLA, had no effect on SCD gene expression and activity. These results suggest that in HepG2 cells the trans-10,cis-12 CLA isomer regulates human SCD activity mainly by a posttranslational mechanism.

Topics: Fatty Acids, Monounsaturated; Humans; Linoleic Acids; Linoleic Acids, Conjugated; Protein Processing, Post-Translational; RNA, Messenger; Stearoyl-CoA Desaturase; Tumor Cells, Cultured

Conjugated linoleic acids (CLA) are octadecadienoic fatty acids that have profound effects on lipid metabolism. Our previous work showed that CLA (mixture of isomers) markedly reduced milk fat synthesis. In this study, our objective was to evaluate the effects of specific CLA isomers. Multiparous Holstein cows were used in a 3x3 Latin square design, and treatments were 4-day abomasal infusions of 1) skim milk (control), 2) 9,11 CLA supplement, and 3) 10,12 CLA supplement. CLA supplements provided 10 g/day of the specific CLA isomer (cis-9,trans-11 or trans-10,cis-12). Treatments had no effect on intake, milk yield, or milk protein yield. Only the 10,12 CLA supplement affected milk fat, causing a 42 and 44% reduction in milk fat percentage and yield, respectively. Milk fat composition revealed that de novo synthesized fatty acids were extensively reduced. Increases in ratios of C(14:0) to C(14:1) and C(18:0) to C(18:1) indicated the 10,12 CLA supplement also altered Delta(9)-desaturase. Treatments had minimal effects on plasma concentrations of glucose, nonesterified fatty acids, insulin, or insulin-like growth factor-I. Overall, results demonstrate that trans-10,cis-12 CLA is the isomer responsible for inhibition of milk fat synthesis.

Topics: Abomasum; Animals; Cattle; Fatty Acids; Female; Injections; Linoleic Acids; Linoleic Acids, Conjugated; Milk; Stereoisomerism

We investigated the cytotoxic effect of conjugated trienoic fatty acids on various human tumor cell lines: DLD- 1, colorectal; HepG2, hepatoma; A549, lung; MCF-7, breast; and MKN-7, stomach. Conjugated linoleic acid (CLA) and conjugated linolenic acid were prepared from linoleic acid (18:2, n-6) and alpha-linolenic acid (18:3, n-3), respectively, by treatment with 6.6% or 21% potassium hydroxide. Spectrophotometric readings at 235 nm for the conjugated diene formation, and at 268 nm for the conjugated triene, were confirmed for the respective conjugated fatty acids. In addition, tung oil (Aleurites fordii) fatty acids consisting principally of a conjugated triene (eleostearic acid, approximately 80% of total fatty acids) were prepared using an alkaline saponification procedure. All tumor cells were incubated for 24 h with 5-100 microM of the conjugated fatty acids, and MTT dye reduction was measured to verify the cell viability. Among the conjugated fatty acids examined, conjugated linolenic acid and tung oil fatty acids exhibited the most intense cytotoxic effects on DLD-1, HepG2, A549, MCF-7 and MKN-7 cells, while CLA was not cytotoxic to the tumor cells. These results demonstrate that conjugated trienoic fatty acids are more cytotoxic to human tumor cells than the conjugated dienoic fatty acid, CLA.

Topics: Antineoplastic Agents; Cell Survival; Coloring Agents; Drug Screening Assays, Antitumor; Humans; Linoleic Acids; Linoleic Acids, Conjugated; Linolenic Acids; Oxidation-Reduction; Plant Oils; Tetrazolium Salts; Tumor Cells, Cultured

Conjugated diene isomers of linoleic acid (CLA) are normal constituents of certain foods and exhibit anticarcinogenic and antiatherogenic properties. In the present study, the effects of several CLA isomers on human platelet aggregation and arachidonic acid metabolism were examined. It was found that 9c,11t-CLA, 10t, 12c-CLA and 13-hydroxy-9c,11t-octadecadienoic acid (13-HODE) inhibited arachidonic acid- and collagen-induced platelet aggregation with I50s in the 5-7 microM range. The nonconjugated 9c, 12c-LA was about 300% and 50%, respectively, less potent an inhibitor with these aggregating agents. Using either thrombin or the calcium ionophore A23187 as aggregating agents, a CLA isomer mix was also found to be more inhibitory than 9c,12c-LA. The 9c,11t- and 10t,12c-CLA isomers as well as the CLA isomer mix inhibited formation of the proaggregatory cyclooxygenase-catalyzed product TXA2, as measured by decreased production of its inactive metabolite [14C]TXB2 from exogenously added [14C]arachidonic acid (I50s=9-16 microM). None of the CLA isomers tested inhibited production of the platelet lipoxygenase metabolite [14C]12-HETE. The additional presence of a hydroxyl group gave opposite results: 13-HODE (I50=3 microM) was about 4-fold more potent a cyclooxygenase inhibitor than the 9c,11t-CLA isomer but 9-HODE was 2- to 3-fold less effective an inhibitor (I50=34 microM) of [14C]TXB2 formation than the corresponding 10t,12c-CLA. In both the aggregation and arachidonic acid metabolism experiments, the inhibitory effects of CLA on platelets were reversible and dependent on the time of addition of either the aggregating agent or the [14C]arachidonic acid substrate. These studies suggest that CLA isomers may also possess antithrombotic properties.

Topics: 12-Hydroxy-5,8,10,14-eicosatetraenoic Acid; Arachidonic Acid; Blood Platelets; Dietary Fats, Unsaturated; Humans; Isomerism; Linoleic Acids; Linoleic Acids, Conjugated; Platelet Aggregation Inhibitors; Thromboxane B2