Adipose depletion and apoptosis induced by trans-10, cis-12 conjugated linoleic Acid in mice

OBJECTIVE: To compare the effectiveness of a conjugated linoleic acid (CLA) isomer mixture (mCLA) with each main isomer [trans-10,cis-12 CLA (CLA10,12) and cis-9,trans-11 CLA (CLA9,11)] in causing body lipid loss and adipose tissue apoptosis. RESEARCH METHODS AND PROCEDURES: Mice selected over 16 generations for high (MH) or low (ML) energy expenditure and a control group (MC) were fed diets containing either soy oil or soy oil plus mCLA, CLA10,12, or CLA9,11 for 5 days in one study and 14 days in a second study. RESULTS: Mice fed mCLA or CLA10,12 had less body lipid (p < 0.05), smaller retroperitoneal fat pads (p < 0.05), and ate less (p < 0.01) than mice fed no CLA or CLA9,11 for 5 days. Mice consuming 1% mCLA or 0.5% CLA10,12 gained less weight (p < 0.01) and had less body lipid (p < 0.05) and smaller epididymal (p < 0.05) and retroperitoneal fat pads (p < 0.01) than mice consuming either control or 0.5% CLA9,11-containing diets for 14 days. Only mCLA and CLA10,12 increased apoptosis in retroperitoneal fat pads (p < 0.01). The effects of mCLA and CLA10,12 were independent of genetic line except for the effect on adipocyte apoptosis. Mice of the MH line were slightly less sensitive than MC or ML mice to CLA-induced adipose tissue apoptosis. DISCUSSION: CLA10,12, but not CLA9,11, can induce both body fat loss and adipose apoptosis. Although mice of a genotype with less body fat and greater metabolic rate and feed intake appear less sensitive, these CLA effects are robust for mice of varying metabolic background.     

Influence of dietary conjugated linoleic Acid and fat source on body fat and apoptosis in mice

OBJECTIVE: To determine whether altered dietary essential fatty acid (linoleic and arachidonic acid) concentrations alter sensitivity to conjugated linoleic acid (CLA)-induced body fat loss or DNA fragmentation. RESEARCH METHODS AND PROCEDURES: Mice were fed diets containing soy oil (control), coconut oil [essential fatty acid deficient (EFAD)], or fish oil (FO) for 42 days, and then diets were supplemented with a mixture of CLA isomers (0.5% of the diet) for 14 days. Body fat index, fat pad and liver weights, DNA fragmentation in adipose tissue, and fatty acid profiles of adipose tissue were determined. RESULTS: The EFAD diet decreased (p < 0.05) linoleic and arachidonic acid in mouse adipose tissue but did not affect body fat. Dietary CLA caused a reduction (p < 0.05) in body fat. Mice fed the EFAD diet and then supplemented with CLA exhibited a greater reduction (p < 0.001) in body fat (20.21% vs. 6.94% in EFAD and EFAD + CLA-fed mice, respectively) compared with mice fed soy oil. Dietary FO decreased linoleic acid and increased arachidonic acid in mouse adipose tissue. Mice fed FO or CLA were leaner (p < 0.05) than control mice. FO + CLA-fed mice did not differ in body fat compared with FO-fed mice. Adipose tissue apoptosis was increased (p < 0.001) in CLA-supplemented mice and was not affected by fat source. DISCUSSION: Reductions in linoleic acid concentration made mice more sensitive to CLA-induced body fat loss only when arachidonic acid concentrations were also reduced. Dietary essential fatty acids did not affect CLA-induced DNA fragmentation.     

Conjugated linoleic acid persistently increases total energy expenditure in AKR/J mice without increasing uncoupling protein gene expression

AKR/J mice fed a high fat diet were treated with a 1% (1 g/100 g) admixture of conjugated linoleic acids (CLA) for 5 wk and compared with control mice. Body weights, energy intakes and energy expenditure (EE) determined by indirect calorimetry were measured weekly. CLA treatment reduced adipose depot weights by approximately 50% but had no significant effects on either body weight or energy intake. CLA increased EE persistently by an average of 7.7% throughout the 5-wk experiment. This greater EE, despite no difference in energy intake, was sufficient to account for the lower body fat stores in the CLA-treated mice. De novo fatty acid biosynthesis in adipose tissue, measured by incorporation of deuterium-labeled water, was not decreased by CLA treatment and therefore did not explain the lower adipose lipid in these mice. Expression of uncoupling protein (UCP) in skeletal muscle, white adipose tissue and kidney was not affected by CLA treatment. In brown adipose tissue, UCP1 expression was not affected by CLA treatment. However, UCP2 expression, although quite low, was significantly greater in CLA-fed mice. We conclude that CLA acts to reduce body fat stores by chronically increasing metabolic rate. This effect on metabolic rate is likely not due to increased UCP gene expression. Furthermore, the reduced body fat is not due to decreased de novo fatty acid synthesis in white adipose tissue.     

Increasing the amount of fat in a conjugated linoleic acid-supplemented diet reduces lipodystrophy in mice

Conjugated linoleic acid (CLA) is a naturally occurring group of dienoic derivatives of linoleic acid found in beef and dairy products. However, when 1 g CLA/100 g diet was given to mice in a low fat diet (4 g fat/100 g diet), they showed a marked decrease in fat mass, but demonstrated symptoms of lipoatrophic diabetes, i.e., marked hepatomegaly and insulin resistance. In this study, to determine whether the decrease in adipose tissue was responsible for these adverse effects, mice were fed different doses of CLA and dietary fat. In Experiment 1, mice were fed different doses of CLA (0, 0.1 and 1 g CLA/100 g diet) in a fixed 4 g fat/100 g diet; in those fed 0.1 g CLA, subcutaneous white adipose tissue (WAT) weight was 48% lower than in mice fed 0 g CLA. The mice fed 0.1 g CLA did not exhibit hepatomegaly and insulin resistance. In Experiment 2, mice were fed for 5 mo different amounts of dietary fat (4, 13 and 34 g fat/100 g diet) in 0 or 1 g CLA/100 g diet; in mice fed 1 g CLA with 34 g fat, retroperitoneal and subcutaneous WAT weights were 76 and 79% lower, respectively, than those of mice fed 0 g CLA with 34 g fat. Mice fed 1 g CLA in the diet with 34 g fat had normal plasma insulin concentrations and a 45% greater liver weight. These data suggested that the percentage of CLA in dietary fat might be a determinant of CLA-mediated lipodystrophy.     

Serum leptin concentrations are not related to dietary patterns but are related to sex, age, body mass index, serum triacylglycerol, serum insulin, and plasma glucose in the US population

Background

The conjugated linoleic acid (CLA) content of beef can be increased by supplementing appropriate beef cattle diets with vegetable oil or oil seed. Yet the effect of consumption of such beef on adipose tissue characteristics is unclear, thus the study was conducted to compare adipose tissue responses of rats to diets containing beef from steers either not provided or provided the oil supplements to alter CLA composition of the fat in muscle.

Methods

Effects of feeding synthetic (industrial hydrogenation) CLA or CLA from beef on growth and adipose tissue responses of weanling, male, Wistar rats (n = 56; 14 per treatment diet) were investigated in a completely randomized design experiment. Diets were: control (CON) diet containing casein and soybean oil, synthetic CLA (SCLA) diet; where 1.69% synthetic CLA replaced soybean oil, two beef-diets; CONM and CLAM, containing freeze dried beef from steers either not fed or fed 14% sunflower seeds to increase CLA content of beef. Diets were isonitrogenous (20% protein) and isocaloric. Rat weights and ad libitum intakes were recorded every 2 wk. After 9 wk, rats were fasted for 24 h, blood sampled by heart puncture, sacrificed, tissue and organs were harvested and weights recorded. The adipose tissue responses with regard to cellularity and fatty acid compositions of retroperitoneal and inguinal adipose tissue were determined.

Results

Body weights and gains were comparable, but organ weights as percent of body weight were greater for rats fed SCLA than CONM. Fasting blood glucose concentration was lower (p < 0.01) in rats fed SCLA than those fed CONM or CLAM. Retroperitoneal and inguinal fat weights, as percent of body weight were greater (p < 0.01) in rats fed CONM or CLAM than those fed CON or SCLA diets. Adipocyte numbers were least in retroperitoneal tissue of rats fed SCLA, while inguinal tissue cell density and total number were lower (p = 0.02) in rats fed CLAM (7.26 × 10⁷ cells/g and 8.03 × 10⁸ cells) than those fed CONM (28.88 × 10⁷ cells/g and 32.05 × 10⁸ cells, respectively).

Conclusion

Study suggests that dietary CLA either as synthetic or high CLA-beef may alter adipose tissue characteristics by decreasing the number of adipocytes and by decreasing the size of the tissue.     

Dietary conjugated linoleic acid reduces rat adipose tissue cell size rather than cell number

We investigated the basis for the reduction in fat pad size in rats fed conjugated linoleic acid (CLA). In the first study, growing female Sprague-Dawley rats (initial weight150 g) were fed diets containing 0, 0.25 and 0.5 g/100 g diet of a purified (97% CLA) and 0.5% of a feed-grade (55% CLA) source of CLA for 5 wk to determine the effects on growth performance and fat mass. There was no effect of CLA on growth rate or food intake. Dietary CLA reduced retroperitoneal fat pad weight 13, 25 and 32% in rats fed 0.25 and 0. 5% of the pure CLA and 0.5% of the feed-grade CLA, respectively (P < 0.05). Similar effects were observed in the parametrial fat pad. The reduced pad size was due to smaller adipocyte size rather than a reduced cell number. Relative to the control group, mean cell volume was 15, 28 and 29% lower in tissue from rats fed 0.25 and 0.5% of the pure CLA and 0.5% of the feed-grade CLA, respectively (P < 0.01). In the second study, rats were fed CLA (0 vs. 0.5%) for 7 or 49 d. Reductions in fat pad weight were observed within 7 d. In addition, the effects of CLA on energy metabolism were studied in the chronically fed rats. There were no significant effects of CLA on oxygen consumption, CO(2) or heat production. During wk 4 of feeding, but not at other times, there was a 5% lower respiratory quotient in CLA-fed rats (P < 0.05). There was a time-dependent accumulation of CLA in adipose tissue and a decrease in monounsaturated fatty acids. These results suggest that the reduction in fat mass in rats fed CLA can be accounted for by a reduction in cell size rather than a change in cell number.     

The combination of dietary conjugated linoleic acid and treadmill exercise lowers gain in body fat mass and enhances lean body mass in high fat-fed male Balb/C mice

Nearly half of the U.S. adult population is overweight or obese, which may be related to increased energy intake combined with lack of physical activity. Obesity increases the risk of several chronic diseases including diabetes, coronary heart disease, hypertension, and stroke. Conjugated linoleic acids (CLA) were shown to decrease fat and increase lean mass in several animal studies. However, the effects of CLA in combination with exercise (Ex) on body composition have not been studied in an animal model. We examined the effect of a low concentration of either safflower oil as control (0.5%) or mixed isomers of CLA (0.4%) along with treadmill exercise on body composition in male Balb/C mice fed a high-fat diet (20% corn oil) in a 2 x 2 factorial design. CLA consumption lowered change in fat mass (P < 0.001) confirming the results of other studies, and change in fat mass decreased further (P < 0.001) with CLA and exercise. Change in lean mass did not increase with exercise alone; it increased, although not significantly, with CLA alone and increased significantly (P < 0.05) due to the combination of CLA and exercise. This effect was accompanied by decreased serum leptin levels and lower leptin mRNA expression in peritoneal fat (P < 0.001). Serum insulin, glucose, tumor necrosis factor (TNF)-alpha, and interleukin-6 were lower in CLA-fed mice than in controls (P < 0.05), whereas serum TNF-alpha was increased by exercise (P < 0.05). Exercise increased oxygen consumption and energy expenditure when measured under resting conditions (P < 0.05). In summary, the combination of dietary CLA and exercise decreased fat mass and increased lean mass in mice fed a high-fat diet, and these effects may be related in part to decreased serum leptin and exercise-induced increases in oxygen consumption and energy expenditure.     

Sensitivity of ob/ob mice to leptin-induced adipose tissue apoptosis

OBJECTIVE: ob/ob mice have increased sensitivity to many of leptin's effects. The primary objective of this experiment was to determine whether ob/ob mice demonstrated increased sensitivity to leptin-induced adipose tissue apoptosis. RESEARCH METHODS AND PROCEDURES: Fifteen-week-old female ob/ob and Ob/? mice received 0 (saline), 2.5, or 10 microg/d leptin for 14 days through subcutaneous (sc) osmotic minipumps. Food intake (FI), body temperature, physical activity, and body weight were measured daily. Body composition and weights and adipose tissue apoptosis (percentage DNA fragmentation) of inguinal, parametrial, and retroperitoneal fat pads were determined at the end of the study. RESULTS: FI decreases were more pronounced in ob/ob. Leptin (10 microg/d) decreased total FI 71% in ob/ob and 34% in Ob/? (p < 0.05). Body weight was decreased by both doses of leptin in ob/ob (p < 0.01) but was unchanged in Ob/?. Leptin increased body temperature in ob/ob but not in Ob/?. Physical activity was increased 400% by 10 microg/d leptin in ob/ob (p < 0.01) but decreased 13% in Ob/? (p < 0.01). Body fat content of ob/ob was reduced by both leptin doses, whereas only 10 microg/d leptin decreased body fat in Ob/?. Fat pad weights were decreased similarly by leptin in both genotypes. However, apoptosis was increased by leptin in all three fat pads in ob/ob, whereas Ob/? showed significant increases only in retroperitoneal. DISCUSSION: ob/ob mice had greater overall sensitivity to leptin. Although ob/ob mice appeared to be more sensitive than Ob/? mice to leptin-induced adipose tissue apoptosis, there were differences among adipose depots in responsiveness to leptin-induced apoptosis.     

Incorporation of conjugated linoleic acid into brain lipids is not necessary for conjugated linoleic acid–induced reductions in feed intake or body fat in mice

Dietary conjugated linoleic acid (CLA) causes reduced feed intake (FI) and body fat (BF). It is unknown, though, if CLA incorporation into tissues, alterations in serum hormones, and/or appetite-regulating neuropeptides are involved. We hypothesized that CLA incorporation into brain lipids would be correlated with changes in appetite-regulating neuropeptide expression and reductions in FI and BF. Male mice (n = 150; 9 weeks old, ICR) received the control diet ad libitum (CON), 2% CLA diet ad libitum (CLA), or control diet pair-fed to the intake of CLA-fed mice for 1, 2, 3, 5, or 7 days. Both FI and body weight were measured daily, and a BF index was calculated. Liver, adipose, and brain fatty acids; serum insulin, leptin, and peptide YY; and arcuate nucleus neuropeptide Y, agouti-related protein, and α-melanocyte–stimulating hormone protein were determined. Mice fed CLA ate less (P < .05) than did the CON on days 1, 2, 3, and 7 but were leaner (P < .05) only on day 7. Mice that received the control diet pair-fed to the intake of CLA-fed mice did not differ in BF from the CON. By days 1 and 2, CLA isomers were incorporated into the liver and adipose but not in the brain. Insulin was increased in CLA-fed mice on days 5 and 7, and leptin was decreased on day 7. Peptide YY and the neuropeptides did not differ. Tissue CLA was not correlated with FI, body weight, or BF but was positively correlated with insulin and negatively correlated with leptin. The reduction in FI is not sufficient to cause the reduction in BF, and tissue CLA accumulation does not appear to be required.     

Dietary conjugated linoleic acid reduces adiposity in lean but not obese Zucker rats

Recent studies have demonstrated a reduction in body fat in growing animals fed conjugated linoleic acid (CLA). Two experiments were conducted to extend these observations to obese rats so that the mechanism of the actions of CLA might be more easily elucidated. In experiment 1, male lean and obese Zucker rats were fed diets containing either 0 or 0.5% CLA for 5 wk. There was no effect of diet on growth rate or food intake. Dietary CLA reduced retroperitoneal and inguinal fat pad weights in the lean rats but increased fat pad weights in the obese genotype (diet x genotype interaction; P < 0.05). Determination of fat pad cellularity indicated that these changes in fat pad weight were due to a reduction or increase in average fat cell size for the lean and obese Zucker rats, respectively. In experiment 2, we sought to reproduce these effects on fat pad size, as well as to determine the effect of dietary CLA on the catabolic response to bacterial endotoxin injection in obese Zucker rats. Growing female lean and obese Zucker rats were fed diets containing 0 or 0.5% CLA for 8 wk. On d 28, each rat was injected intraperitoneally with lipopolysaccharide from Escherichia coli serotype 055:B5 (1 mg/kg body weight) and body weight was determined over the next 96 h. There was a diet x genotype interaction (P < 0.05) for the body weight response to lipopolysaccharide 24 h postinjection. Lean rats fed CLA lost less weight than did lean controls, but obese rats fed CLA lost more weight than did obese controls. As in the first experiment, there was a diet x genotype (P < 0.05) for the effect of treatment on retroperitoneal fat pad weights determined at the end of the experiment. Lean rats fed CLA had smaller RP fat pads than did lean controls, but obese rats fed CLA once again had heavier RP fat pads than did obese controls. These results indicate that CLA reduces body fat and catabolic response to endotoxin injection in lean Zucker rats but not in the obese genotype. The observed interaction between diet and genotype warrants additional investigation into the specific mechanism(s) of the biological activities of CLA.     

Trans-vaccenic acid is desaturated to conjugated linoleic acid in mice

Mice were fed pure trans11 octadecenoic acid (trans-vaccenic acid; TVA) to determine whether it is desaturated to cis9, trans11 octadecadienoic acid, a predominant isomer of conjugated linoleic acid (CLA). In a preliminary trial, 12% of the TVA consumed during a 2-wk feeding period was recovered in the carcass as CLA. As a proportion of TVA in the tissues available for bioconversion, 48.8% was desaturated. We tested whether desaturation could be modified by supplementing no modifier, 0.5% clofibric acid to stimulate desaturation, or increasing the polyunsaturated fatty acids (PUFA) (10% corn oil vs. 4% corn oil) to inhibit desaturation in diets with or without 1% TVA. These diets were fed to six groups of mice in a 3x2 factorial arrangement of treatments. Feeding 1% TVA with 10% corn oil decreased feed intake (2.70 vs. 3.73 g/d, SEM 0.23; P<0.05). Bioconversion of dietary TVA was 12.0, 7.5 and 5.1% for mice fed no modifier of desaturation, clofibrate and increased PUFA, respectively. Conversion based on TVA available for desaturation was 52.6, 55.5 and 37.0%, respectively. Thus, clofibrate did not increase bioconversion, but increasing PUFA decreased conversion by 30%. To test whether TVA decreases food intake directly or after conversion to CLA, four groups of mice were fed diets containing 1% stearic, TVA, elaidic or conjugated linoleic acid. Dietary CLA decreased food intake and body fat, but did not change body protein. CLA was found in the carcass only when TVA or CLA was fed. CLA was found in both triacylglycerol and phospholipids when CLA was fed, but only in triacylglycerol when TVA was fed, suggesting that bioconversion occurred in the adipose tissue. In three trials, conversion of dietary TVA to CLA was 11.4+/-1.25%; conversion of stored TVA was 50.8+/-1.91%. Similar bioconversion of TVA in humans would increase current estimates of CLA available for the general population by 6- to 10-fold.     

Conjugated linoleic acid induces mast cell recruitment during mouse mammary gland stromal remodeling

Conjugated linoleic acid (CLA) is a dietary chemopreventive agent that induces apoptosis in the mammary adipose vascular endothelium and decreases mammary brown adipose tissue (BAT) and white adipose tissue (WAT). To determine onset and extent of stromal remodeling, we fed CD2F1/Cr mice diets supplemented with 1 or 2 g/100 g mixed CLA isomers for 1-7 wk. BAT loss, collagen deposition, and leukocyte recruitment occurred in the mouse mammary fat pad, coincident with an increase in parenchymal-associated mast cells in mice fed both levels of CLA. Feeding experiments with purified isomers (0.5 g/100 g diet) demonstrated that these changes were induced by trans-10, cis-12 CLA (10,12-CLA), but not by cis-9, trans-11 CLA (9,11-CLA). This stromal remodeling did not require tumor necrosis factor (TNF)-alpha, a major cytokine in mast cells, as TNF-alpha null mice demonstrated collagen deposition, increased leukocytes, and BAT loss in the mammary fat pad in response to 10,12-CLA. To test the hypothesis that mast cells recruited in response to 10,12-CLA were required for stromal remodeling, Steel mice (WBB6F1/J-kit(W)/kit(W-V)), which lack functional mast cells, were examined for their stromal response to 10,12-CLA. Both wild-type and Steel mice showed a significantly increased leukocytic adipose infiltrate, collagen deposition, and decreased adipocyte size, although BAT was maintained in Steel mice. These results demonstrate that 10,12-CLA induces an inflammatory and fibrotic phenotype in the mouse mammary gland stroma that is independent of TNF-alpha or mast cells and suggest caution in the use of 10,12-CLA for breast cancer chemoprevention.     

Dietary conjugated linoleic acid affects morphofunctional and chemical aspects of subcutaneous adipose tissue in heavy pigs

We investigated the in vivo effects of dietary conjugated linoleic acid (CLA) on subcutaneous adipose tissue from heavy pigs to clarify the involvement of possibly different causative effects in the established antiadipogenic effect of CLA. Pigs [n = 36; initial body weight, 106 kg live weight (LW)] were assigned to 1 of 2 LW-matched groups supplemented with either 0 or 0.75% of a CLA preparation containing 50% CLA isomers. The pigs were slaughtered at 155 kg LW and adipose tissue analyzed. CLA supplementation affected ash content, and decreased iodine values (P < 0.01) and adipocyte size (P < 0.05). The fat content of adipose tissue was lower (P < 0.05) in females than castrated males, and females had smaller (P < 0.01) adipocytes than castrated males. Neither CLA nor sex influenced adipocyte lipid droplet diameter or the extent of lipid peroxidation as determined by quantitation of Schiff's histochemical reaction. NADPH-diaphorase was not influenced by CLA treatment. Preadipocyte proliferation rates were lower in pigs fed CLA (P < 0.05), whereas the number of adipocyte apoptotic nuclei was greater (P < 0.05). Preadipocyte proliferation was also greater (P < 0.05) in females than castrated males. Neuronal and endothelial nitric oxide synthase activities did not differ between groups in adipose tissue vessels, but inducible nitric oxide synthase expression in adipocytes was lower in pigs fed CLA (P = 0.05). These findings suggest that the antiadipogenic effect of CLA in heavy pigs is not a direct effect but may occur by downregulation of a NO-mediated lipolytic pathway.     

Effects of conjugated linoleic acid on growth performance, feed conversion efficiency, and subsequent carcass quality in broiler chickens

The effect of dietary conjugated linoleic acid isomers (CLA) on growth performance, carcass composition, fatty acid composition of adipose and muscle tissues, and serum lipoproteins was investigated in broiler chickens. A total of 160 (eighty male and eighty female) chickens were allocated to four dietary treatments (0.0, 0.5, 1.0, and 1.5 % CLA) and fed a standard starter diet from 8 to 21 d, and a grower-finisher diet from 22-42 d. When determined for the total period 8-42 d, feed intake and body weight gains of broiler chickens were significantly reduced (from 3.31 to 3.12 kg and from 1615 to 1435 g respectively; P < 0.05), particularly at the 1.5 % dietary CLA level. Feed conversion efficiency and carcass yield values showed no significant effects of dietary CLA. Abdominal fat deposition was significantly reduced (from 2.68 to 1.78 %; P < 0.05), the relative proportion of breast muscles was unaffected, and that of leg muscles significantly increased (from 19.0 to 20.6 %; P < 0.05). The concentration of CLA isomers (% of total methyl esters of fatty acids) increased linearly in tissue samples from broilers fed 0.5, 1.0, and 1.5 % dietary CLA. The relative proportions of saturated fatty acids (16:0, 18:0) were significantly (P < 0.01) increased, and those of monounsaturated (16:1, 18:1) and polyunsaturated fatty acids (18:2, 20:4 in muscle tissues) significantly (P < 0.05) reduced. Total serum cholesterol concentrations reached a maximum in broilers fed 1.0 % CLA and then decreased slightly (from 141.73 to 136.47 mg/dl; P < 0.01). The same was true also for HDL-cholesterol (from 113.58 to 109.97 mg/dl; The HDL cholesterol:total cholesterol ratio and serum triacylglycerol concentration was unaffected. In conclusion, feeding CLA to broiler chickens resulted in substantial incorporation of CLA isomers into their tissue lipids, thus providing a potential CLA-rich source for human consumption.     

Dietary conjugated linoleic acid consumption during pregnancy and lactation influences growth and tissue composition in weaned pigs

We evaluated the effects of conjugated linoleic acid (CLA) on growth performance, tissue fatty acid composition and ex vivo lipogenic enzyme activity in piglets (n = 40) reared on sows fed diets supplemented with CLA or linoleic acid (LA). Weaned offspring of both sow groups were offered either a CLA- or LA-enriched starter diet for 35 d. The starter diets were formulated to contain 2 g CLA (containing 58.9 g CLA/100 g total fatty acids) or LA per 100 g feed. All piglets were slaughtered at 70 d of age and tissue samples of the back fat, omental fat and longissimus dorsi were collected. Irrespective of the dietary fat supplied in the starter period, piglets reared on the CLA sows had greater final body and warm carcass weights (P: < 0.01), and greater feed intake (P: = 0.02) than piglets reared on the LA sows. The dietary effect on the fatty acid composition was similar for the adipose and muscle tissues. Compared with the LA-enriched diets, CLA increased the level of total saturated fatty acids (P: < 0.05), whereas that of monounsaturated fatty acids was decreased (P: < 0.05). Dietary CLA increased glucose-6-phosphate dehydrogenase (P: < 0.01) and malic enzyme activities (P: < 0.06) in the fat tissues, but did not affect fatty acid synthase activity. The shift toward a higher deposition of saturated fatty acids and a lower deposition of monounsaturated fatty acids is the result of down-regulation of Delta9-desaturase activity that was induced by CLA rather than an altered rate of de novo synthesis.     

The decrease in body fat in mice fed conjugated linoleic acid is due to increases in energy expenditure and energy loss in the excreta

We carried out energy balance studies in four groups of young, growing, 5-wk-old Balb-C mice (n = 12/group) that were either food restricted or nonrestricted and fed high fat diets (38 energy%) with or without 0.93 g/100 g conjugated linoleic acid (CLA) for 39 d. The energy in carcasses, excreta and food was measured in a bomb calorimeter. CLA lowered the percentage of the energy intake that was stored in the body from 1.9 +/- 0.8 to -2.3 +/- 0.7% (mean +/- SD, P < 0.05) in the nonrestricted mice and from 1.4 +/- 1.3 to -2.9 +/- 0.7% (P < 0.05) in the restricted mice. Thus, the CLA-treated mice had a net loss of body energy. The percentage of the energy intake eliminated in the excreta increased from 7.6 +/- 0.9% in controls to 8.7 +/- 1.0% (P < 0.05) in the CLA-treated mice that were nonrestricted and from 7.3 +/- 0.8 to 8.4 +/- 0.6 (P < 0.05) in the restricted mice. The amount of energy ingested minus the amount retained in carcasses and excreta equals the energy expenditure. The percentage of the energy intake that was expended as heat increased from 90.5 +/- 1.2 in controls to 93.6 +/- 1.5% (P < 0.05) in the CLA-treated nonrestricted mice and from 91.3 +/- 1.5 to 94.5 +/- 1.0% (P < 0.05) in the restricted mice. The lower energy storage in the CLA-fed mice was accounted for by an increase in the energy expenditure (74%) and by an increase in energy lost in the excreta (26%). Feeding CLA also increased liver weight, which may warrant further studies on the safety of CLA.     

Changes in Body Composition with Conjugated Linoleic Acid

Conjugated linoleic acid has been shown to reduce body fat accumulation in several animal models. We have conducted several studies in AKR/J mice showing that CLA reduces body fat accumulation whether animals are fed a high-fat or low-fat diet, with no effect on food intake. One mechanism by which CLA reduces body fat is by increased energy expenditure, which is observed within one week of CLA feeding and is sustained for at least six weeks. The increased energy expenditure is sufficient to account for the decreased fat accumulation. Increased uncoupling protein gene expression does not appear to be involved in the increased energy expenditure. We have observed increased fat oxidation but no decrease in de novo fat biosynthesis with CLA feeding. We have also observed increased liver weights and plasma insulin levels with higher doses of CLA. In all of the studies we have conducted to date we have used a CLA preparation that contains several isomers, primarily c9,t11 and t10,c12. It was assumed that the active form was c9,t11, as CLA was identified as an anticarcinogenic compound from cooked beef, of which the c9,t11 form accounts for 60% to 80% of the CLA. Most of the studies conducted so far must be repeated using the purified isomers in order to determine which isomers are responsible for each of the identified actions of CLA.     

Genistein decreases food intake, body weight, and fat pad weight and causes adipose tissue apoptosis in ovariectomized female mice

Genistein, an isoflavone in soybean products, has estrogenic activity and is used as a natural substitute for estrogen replacement therapy in postmenopausal women. Genistein was also shown to decrease fat pad weight in female mice. The primary objective of this study was to determine the effect of genistein on adipose tissue apoptosis in vitro and in vivo. 3T3-L1 preadipocytes and mature adipocytes were treated with 0, 1, 10, 100, and 400 micromol/L genistein and then assayed for apoptosis, whereas only mature adipocytes were assayed for viability. Mature adipocytes treated with genistein demonstrated a dose-related increase in apoptosis. Ovariectomized female mice (9 mo old) were given 0, 150, or 1,500 mg/kg genistein in the semipurified phytoestrogen-free casein-based diet for 3 wk (n=10). After mice were killed, body composition was determined by dual-energy X-ray absorptiometry analysis, and parametrial (PM), inguinal (ING), and retroperitoneal (RP) fat pads were weighed and assayed for apoptosis (% DNA fragmentation). Genistein (1500 mg/kg) reduced food intake (FI) by 14% (P<0.01) and body weight (BW) by 9% (P<0.01). Body composition was not significantly affected, but PM and ING weights were decreased 22% (P<0.05) and 19% (P<0.07), respectively, by 1,500 mg/kg genistein. Apoptosis in ING fat was increased 290% (P<0.05) by 1,500 mg/kg genistein. These findings show that oral genistein treatment can reduce BW, mobilize body fat, and induce apoptosis of adipose tissue in ovariectomized female mice. Thus, genistein may be useful in treating or preventing increased adiposity after menopause.     

Pre- and postnatal dietary conjugated linoleic acid alters adipose development, body weight gain and body composition in Sprague-Dawley rats

Sprague-Dawley rats were fed either a control diet (7 g/100 g soybean oil) or a conjugated linoleic acid (CLA) diet (6.5 g/100 g soybean oil and 0.5 g/100 g CLA) beginning on d 7 of gestation to determine whether pre- and postnatal CLA affects short- and long-term growth and adiposity. At weaning (d 21), progeny were assigned control or CLA diet and fed until 11 wk of age. At birth, litter size and weight were not different between treatments. There were age- and sex-dependent changes in inguinal adipose fatty acid composition at birth and weaning, whereas there were no differences in lipid accretion or adipocyte proliferation. At weaning, CLA did not alter inguinal adipocyte proliferation but increased (P < 0.01) CCAAT/enhancer binding protein alpha expression in inguinal adipose tissue from females, whereas there was no difference in expression in males. Significant differences in size distribution of inguinal adipocytes at weaning and retroperitoneal adipocytes at 11 wk of age were observed. In general, CLA increased the proportion of smaller cells and decreased the proportion of larger cells. The main long-term effect of the dams' diet was the significantly heavier gastrocnemius and soleus muscles, and significantly longer tail lengths, an indication of skeletal growth, of male pups whose dams were fed CLA. Postweaning diet reduced fat pad weights in female but not male pups fed CLA. This response was due to differences in cell size rather than number. Response to CLA treatment may depend on the sex and age of the animal as well as duration of feeding.     

Short-term intake of conjugated linoleic acid inhibits lipoprotein lipase and glucose metabolism but does not enhance lipolysis in mouse adipose tissue

Feeding diets supplemented with t10c12 conjugated linoleic acid (CLA) to growing mice reduces body fat mass. The effects are evident after 1 wk and maximal by 3 wk and are accompanied by reductions in fat cell size. This may complicate direct comparisons with adipocytes from control mice. Accordingly, we investigated the early biochemical events that occur within adipocytes during the first week of CLA feeding, before changes in the size of adipocytes have occurred. Female ICR mice were fed a control diet or a diet supplemented with 0.5 g/100 g of CLA for 4 d, at which time there were no differences in body weight, fat mass or adipocyte size (except that CLA-fed mice had fewer adipocytes >90 micro m in diameter). Parametrial adipose tissue from the CLA-fed mice had significantly reduced heparin-releasable lipoprotein lipase (LPL) and intracellular LPL activities and significantly reduced glucose incorporation into CO(2), fatty acid and glycerol. There were no differences between adipose tissues from CLA-fed or control mice in the ratios of 16:0 to 16:1 and 18:0 to 18:1 fatty acids or in norepinephrine-stimulated lipolysis. Serum insulin levels in food-deprived mice, measured at 4 d and 7 wk, did not differ between groups nor did the concentration of free fatty acids in serum of food-deprived or fed mice measured at the same time points. In mice, CLA-induced inhibition of heparin-releasable LPL and glucose metabolism may be the most important early steps leading to subsequent body fat reduction. In addition, CLA does not appear to enhance lipolysis in mouse adipose tissue in vivo.