The role of lipid in the management of methylmalonic acidaemia: administration of linoleic acid does not increase excretion of methylmalonic acid.
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We have previously shown that a diet containing a mixture of conjugated linoleic acid (CLA) isomers reduces the incidence of colon tumors in rats treated with 1,2-dimethylhydrazine (DMH). The present study examined which of the two main CLA isomers, trans-10,cis-12 CLA (t10c12) or cis-9,trans-11 CLA (c9t11), decreases colon tumor numbers and the mechanisms for this effect. Six-week-old, male Sprague-Dawley rats were intramuscularly injected with 15 mg/kg of DMH twice per week for 6 weeks and fed a control diet, 1% t10c12, or 1% c9t11 for 30 weeks. The experimental diets were initiated simultaneously with DMH injection. The tumor numbers were decreased and the apoptotic index was significantly increased in the colonic mucosa of the t10c12 and c9t11 groups, when the results were compared with those of the control group. The protein levels of Bcl-2 and cyclooxygenase-2 were significantly decreased, but Bax levels were increased in both of the CLA isomer groups. The thromboxane B(2) levels in colonic mucosa were substantially lower in the two CLA isomer groups than in the control group. However, there was no difference in these parameters between the CLA isomer groups. We have demonstrated that diets containing 1% t10c12 and c9t11 were equally effective in reducing tumor numbers and inducing apoptosis in the colonic mucosa of rats treated with DMH. These results indicate that Bcl-2 family protein levels are associated with CLA-induced apoptosis in the colonic mucosa of DMH-treated rats.
BACKGROUND: High-fat dairy foods contain many potentially anticarcinogenic factors, including conjugated linoleic acid (CLA). However, few epidemiologic studies have specifically evaluated high-fat dairy food consumption, and none have evaluated CLA intake, in relation to colorectal cancer risk. OBJECTIVE: The aim of this study was to prospectively examine the associations of long-term high-fat dairy food consumption and CLA intake and the incidence of colorectal cancer. DESIGN: Our study population consisted of 60 708 women aged 40-76 y who participated in the Swedish Mammography Cohort. The women's consumption of high-fat dairy foods was assessed at baseline, which was from 1987 to 1990, and again in 1997. RESULTS: We ascertained 798 incident cases of colorectal cancer during an average 14.8 y of follow-up. After adjustment for age and other potential confounders, the women who consumed > or =4 servings of high-fat dairy foods/d (including whole milk, full-fat cultured milk, cheese, cream, sour cream, and butter) had a multivariate rate ratio of colorectal cancer of 0.59 (95% CI: 0.44, 0.79; P for trend = 0.002) when compared with the women who consumed <1 serving/d. Each increment of 2 servings of high-fat dairy foods/d corresponded to a 13% reduction in the risk of colorectal cancer (multivariate rate ratio: 0.87; 95% CI: 0.78, 0.96). For CLA, the multivariate rate ratio of colorectal cancer in a comparison of the 2 extreme quartiles of intake was 0.71 (95% CI: 0.55, 0.91; P for trend = 0.004). CONCLUSION: These prospective data suggest that high intakes of high-fat dairy foods and CLA may reduce the risk of colorectal cancer.
Apoptotic pathways in breast cancer cells are frequently altered, reducing the efficiency of radiotherapy. Conjugated linoleic acid (CLA), known to trigger apoptosis, was tested as radiosensitizer in breast cancer cells MCF-7 and MDA-MB-231. The CLA-mix, made up of the isomers CLA-9cis 11trans and CLA-10trans 12cis, was compared to three purified isomers, i.e., the CLA-9cis 11cis, CLA-9cis 11trans, and CLA-10trans 12cis. Using the apoptotic marker YO-PRO-1, the CLA-9cis 11cis at 50 micro mol/L turned out to be the best apoptotic inducer leading to a 10-fold increase in MCF-7 cells and a 2,5-fold increase in MDA-MB-231 cells, comparatively to the CLA-mix. Contrary to previous studies on colorectal and prostate cancer cells, CLA-10trans 12cis does not lead to an apoptotic response on breast cancer cell lines MCF-7 and MDA-MB-231. Our results also suggest that the main components of the CLA-mix (CLA-9cis 11trans and CLA-10trans 12cis) are not involved in the induction of apoptosis in the breast cancer cells studied. A dose of 5 Gy did not induce apoptosis in MCF-7 and MDA-MB-231 cells. The addition of CLA-9cis 11cis or CLA-mix has allowed us to observe a radiation-induced apoptosis, with the CLA-9cis 11cis being about 8-fold better than the CLA-mix. CLA-9cis 11cis turned out to be the best radiosensitizer, although the isomers CLA-9cis 11trans and CLA-10trans 12cis have also reduced the cell survival following irradiation, but using a mechanism not related to apoptosis. In conclusion, the radiosensitizing property of CLA-9cis 11cis supports its potential as an agent to improve radiotherapy against breast carcinoma.
Effects of conjugated linoleic acids (CLA) on a series of metabolic events are expected to depend on the feeding regimen and levels of energy ingested. This study was the first examining the mode of action of CLA on body composition, tissue lipids, lipoproteins and hepatic enzymes in situations of enhanced fat store mobilization. Two groups of male growing Sprague-Dawley rats were fed for 3 wk a diet containing 0 (control group) or 3 g/100 g of a CLA mixture at the expense of sunflower oil, and were then subjected to a weight-loss feeding regimen for another 18 d. Rats fed the CLA-fortified diet gained 11% less weight than the control rats (P<0.05). Rats fed the high CLA diet had less body fat (1.47+/-0.16 vs. 1.07+/-0.09 g/100g, P<0.05) and a higher lean deposition (25.6+/-0.2 vs. 28.4+/-0.3 g/100 g, P<0.05) than control rats. CLA-fed rats had a 41% lower cholesterol concentration in liver than the control rats (P<0.05). Some differences in glycerophospholipid subclass profile of liver and erythrocyte membrane were observed; the hepatic concentrations of phosphatidylethanolamine (4.76+/-0.46 vs. 6.86+/-0.99 micromol/g, P = 0.07) and phosphatidylcholine (12.9+/-0.5 vs. 15.3+/-1.2 micromol/g, P = 0.09) tended to be greater and the level of phosphatidylcholine in erythrocyte membranes was significantly greater (1.40+/-0.12 vs. 1.83 +/-0.16 micromol/g, P<0.05) in the CLA-treated group than in the control group. The activities of catalase and ornithine decarboxylase in liver did not differ between the groups. Further, CLA-treated rats had significantly lower serum concentrations of VLDL lipids than control rats, whereas concentrations of LDL and HDL lipids were unaffected. The results indicate that a high dose of a CLA mixture is a strong repartitioning agent and a modulator of lipid metabolism under conditions of enhanced fat store mobilization in rats.
BACKGROUND: The aim of this study was to evaluate the effect of linoleic acid (LA) and gamma-linolenic acid (GLA), both precursors of PGE(1), on tear production, tear fluorescein clearance and on the ocular surface after photorefractive keratectomy (PRK). METHODS: Sixty subjects (age 25+/-10 years; refractive error -3+/-2 diopters (spherical equivalent), mean +/- standard deviation) undergoing PRK were enrolled. The inclusion criteria were: Schirmer 1 test >10 mm/5 min, no corneal fluorescein staining, low irritation symptoms (questionnaire score <5), standardised visual scale (to evaluate tear fluorescein clearance) score <3. Patients were randomly divided into two groups. One group of 31 subjects was treated once daily orally with tablets containing LA (28.5 mg) and GLA (15.1 mg) (from 3 days before PRK to 1 month after PRK). The control group (29 subjects) underwent PRK and received no treatment with LA and GLA. A symptoms questionnaire, Schirmer 1 test and fluorescein clearance test (FCT) using the standardised visual scale were performed before starting therapy (T(0)) and 30 days after PRK (T(1)). RESULTS: All patients completed the study. The Schirmer 1 test varied from 16.3+/-6.9 (T(0)) to 17.6+/-7.2 (T(1)) for the treated group and from 18.3+/-6.2 (T(0)) to 15.7+/-7.4 (T(1)) for the untreated group ( P<0.0001, two-tailed unpaired t-test). FCT was 1.9+/-0.6 at T(0) and 1.6+/-0.8 at T(1) for the treated group and 1.7+/-0.7 at T(0) and 2.0+/-0.9 at T(1) for the untreated group ( P<0.0001). The symptoms score was 4.7+/-1.9 at T(0) and 7.6+/-7.2 at T(1) for the treated group and 4.2+/-2.0 at T(0) and 10.1+/-7.6 at T(1) for the untreated group ( P<0.05). CONCLUSION: Reduced corneal sensitivity has already been proved after PRK. This could be the main reason for a decrease in tear production and for a reduced blinking rate leading to delayed tear clearance. The oral precursors of PGE(1), LA and GLA, could be helpful in increasing tear production and clearance after PRK.
Both 2,3,7,8-Tetrachlorodibenzo-p-dioxin (TCDD) and carbon tetrachloride (CCl4) have conspicuous effects on lipid metabolism in rat liver. Although it is generally accepted that CCl4 administration leads to hepatic lipid peroxidation in vivo, conflicting reports from different laboratories make it unclear whether or not lipid peroxidation is involved in the mechanism of toxicity of TCDD. The present study involved pretreating F344 rats with CCl4 or TCDD, then at predetermined times thereafter, giving [U-14C]linoleic acid. A variety of compound classes were monitored in extracts of liver taken 30 min after the label was given. A previously unreported effect of CCl4 was a conspicuous increase in turnover of 1,2-diglycerides. That CCl4 did cause lipid peroxidation was evident from the presence of allylic hydroxyacids not seen in vehicle-treated controls, greatly increased radioactivity in protein-bound material, and decreased levels of arachidonate without decreased synthesis from linolate. Where effects of TCDD pretreatment could be seen, they were much less than the corresponding effects of CCl4. No allylic hydroxyacids were detected in livers of TCDD-treated rats. The concentration of arachidonate was not reduced, and elongation of linolate was not stimulated, indicating that TCDD did not cause extensive-but-repaired peroxidation. It is concluded that while TCDD may slightly increase hepatic lipid peroxidation in rats in vivo, the extent of such stimulation appears to be too slight to account for the toxicity of TCDD.
Recent studies showed that conjugated linoleic acids (CLA) lower triacylglycerol concentrations in the milk of lactating animals. This study was performed to determine the reasons for this phenomenon; we also investigated whether there is a relation between altered lipid metabolism in the liver and the reduction in milk triacylglycerols in rats fed CLA. Two groups of female rats were fed diets containing 0 [sunflower oil (SFO) group] or 14.7 g/kg diet of a CLA mixture (CLA group) at the expense of sunflower oil during growth, pregnancy, and lactation. CLA-fed rats had 49 and 80% lower mRNA concentration and activity of fatty acid synthase, respectively, a 51% lower mRNA concentration of lipoprotein lipase (LPL) in their mammary glands at d 17 of lactation, and a 46% lower milk fat content than SFO rats (P < 0.05). Although CLA rats had lower concentrations of triacylglycerols in the liver than SFO rats (20.8 +/- 2.6 vs. 62.6 +/- 27.7 micromol/g, P < 0.05), concentrations of triglycerides in plasma, which are the substrates of LPL, did not differ between the groups. Moreover, the number of pups per litter, litter weights, and pup weights at d 17 of lactation were 41, 35, and 22% lower, respectively, in the CLA group than in the SFO group. In conclusion, the present study suggests that dietary CLA reduces triacylglycerol concentrations in the milk via reduced de novo fatty acid synthesis in the mammary gland and an impaired uptake of fatty acids from lipoproteins into the mammary gland. This might be the reason for reduced growth rates and an increased mortality of suckling pups.
This study was conducted to examine the effects of dietary supplementation with vegetable oils on performance of high-yielding lactating cows and conjugated linoleic acid (CLA) content in milk fat. Twelve lactating Holstein cows in early lactation (30 to 45 d postpartum) were used in a triple 4 x 4 Latin square design. In each period, the cows in each group were fed the same basal diet and received one of the following treatments: 1) control (without oil), 2) 500 g of cottonseed oil, 3) 500 g of soybean oil, and 4) 500 g of corn oil. Each experimental period lasted for 3 wk, with the first 2 wk used for adaptation to the diet. Supplementation with vegetable oils tended to increase milk yield, with the highest milk yield in the cottonseed oil group (35.0 kg/d), compared with the control (34.4 kg/d). Milk fat percentage was decreased, but there were few effects on percentage and yield of milk protein as well as milk fat yield. The cows fed added soybean oil produced milk with the highest content of trans-11 C(18:1) (23.8 mg/g of fat), which was twice that of the control (12.6 mg/g of fat). Content of cis-9, trans-11 CLA in milk fat increased from 3.5 mg/g in the control to 6.0, 7.1, and 10.3 mg/g for the cows fed oils from cottonseed, corn, and soybean, respectively. A significant linear relationship existed between trans-11 C(18:1) and cis-9, trans-11 CLA. Supplementation with oils doubled the content of total fatty acids in blood plasma, with little difference between different vegetable oil sources. Octadecenoic acid content was significantly higher in blood plasma of animals fed added oils from cottonseed and soybean than those fed with corn oil and control. The plasma trans-11 C(18:1) content was significantly higher in the oil-added animals than in control. Supplementation of vegetable oils tended to improve milk production of lactating cows, and the CLA content in milk fat was significantly increased. Soybean oil seemed to be the optimal source to increase CLA production.
AIM: The aim of this study was to evaluate the accruing effects of conjugated linoleic acid (CLA) supplementation and aerobic exercise (Ex) on body composition and serum lipid profile in humans. METHODS: Forty-four healthy female young subjects were divided ExCLA, CLA, Ex and control (C) groups. The groups of CLA and ExCLA were supplemented with 3.6 g/d CLA whilst ExCLA and Ex groups were exercised for 30 min(-1.)3 days(-1.)week(-1) for 6 weeks. RESULTS: After the study period, fat ratio, fat mass, waist and hip girths were reduced in all experimental groups and fat-free mass induced in ExCLA and CLA groups and body weight was reduced in the CLA group when compared to baseline levels. These alterations were significantly different than those of controls with the highest variations were observed in the ExCLA. There was no change in serum leptin, apo-AI, apo-B, total cholesterol, HDL, LDL, free fatty acid, and triglyceride levels. Serum glucose concentrations of ExCLA and CLA groups and insulin level of ExCLA group decreased significantly as compared to baseline levels with only serum glucose reduction of both groups were significantly different than those of controls (P<0.05). Endurance performance significantly increased in ExCLA and Ex groups (P<0.01) but did not vary in the CLA and controls. CONCLUSIONS: It was shown that both CLA and exercise were effective in improvement of body composition and these effects were cumulated when they have been used together. CLA supplementation alone or with exercise seems effective on serum glucose and insulin concentrations but ineffective on endurance performance.
Isomers of conjugated linoleic acid (CLA) are found in beef, lamb and dairy products. Diets containing CLA reduce adipose mass in various depots of experimental animals. In addition, CLA delays the onset of diabetes in the ZDF rat model for obesity-linked type 2 diabetes mellitus. We hypothesize that there would be an inverse association of CLA with body weight and serum leptin in subjects with type 2 diabetes mellitus. In this double-blind study, subjects with type 2 diabetes mellitus were randomized into one of two groups receiving either a supplement containing mixed CLA isomers (CLA-mix; 8.0 g daily, 76% pure CLA; n = 12) or a supplement containing safflower oil (placebo; 8.0 g daily safflower oil, n = 9) for 8 wk. The isomers of CLA in the CLA-mix supplement were primarily c9t11-CLA ( approximately 37%) and t10c12-CLA ( approximately 39%) in free fatty acid form. Plasma levels of CLA were inversely associated with body weight (P < 0.05) and serum leptin levels (P < 0.05). When levels of plasma t10c12-CLA isomer were correlated with changes in body weight or serum leptin, t10c12-CLA, but not c9t11-CLA, was inversely associated with body weights (P < 0.05) and serum leptin (P < 0.02). These findings strongly suggest that the t10c12-CLA isomer may be the bioactive isomer of CLA to influence the body weight changes observed in subjects with type 2 diabetes. Future studies are needed to determine a causal relationship, if any, of t10c12-CLA or c9t11-CLA to modulate body weight and composition in subjects with type 2 diabetes. Furthermore, determining the ability of CLA isomers to influence glucose and lipid metabolism as well as markers of insulin sensitivity is imperative to understanding the role of CLA to aid in the management of type 2 diabetes and other related conditions of insulin resistance.
OBJECTIVES: To study the effects of c9,t11-conjugated linoleic acid (c9,t11-CLA) on invasive ability of human gastric carcinoma cell line (SGC-7901) and to explore its possible mechanism. METHODS: Reconstituted basement membrane invasion assay was used to evaluate invasive ability of cancer cells. Expression of TIMP-1, TIMP-2 and nm23-H(1) mRNA was measured by reverse transcription polymerase chain reaction (RT-PCR) in SGC-7901 cells. RESULTS: At the concentrations of 200 micromol/L, 100 micromol/L and 50 micromol/L, c9,t11-CLA suppressed their reconstituted basement membrane invasion of SGC-7901 by 53.7%, 40.9% and 29.3%, respectively. c9,t11-CLA could induce the expression of TIMP-1, TIMP-2 and nm23-H(1) mRNA in SGC-7901 cells. CONCLUSIONS: The invasion of SGC-7901 cells could be inhibited by c9,t11-CLA through reconstituted basement membrane. Anti-invasion action of c9,t11-CLA might be associated with induction of expression of TIMP-1, TIMP-2 and nm23-H(1) mRNA in tumor cells.
OBJECTIVE: To study the effects of 13 weeks conjugated linoleic acid (CLA) supplementation in overweight subjects after weight loss on weight regain, body composition, resting metabolic rate, substrate oxidation, and blood plasma parameters. DESIGN: This study had a double-blind, placebo-controlled randomized design. Subjects were first submitted to a very-low-calorie diet (VLCD 2.1 MJ/d) for 3 weeks after which they started with the 13-week intervention period. They either received 1.8 g CLA or placebo per day (low dosage, LD) or 3.6 g CLA or placebo per day (high dosage, HD). SUBJECTS: A total of 26 men and 28 women (age 37.8+/-7.7 y; body mass index (BMI) 27.8+/-1.5 kg/m(2)). MEASUREMENTS: Before VLCD (t=-3), after VLCD but before CLA or placebo intervention (t=0) and after 13-week CLA or placebo intervention (t=13), body weight, body composition (hydrodensitometry and deuterium dilution), resting metabolic rate, substrate oxidation, physical activity, and blood plasma parameters (glucose, insulin, triacylglycerol, free fatty acids, glycerol and beta-hydroxy butyrate) were measured. RESULTS: The VLCD significantly lowered body weight (6.9+/-1.7%), %body fat, fat mass, fat-free mass, resting metabolic rate, respiratory quotient and plasma glucose, insulin, and triacylglycerol concentrations, while free fatty acids, glycerol and beta-hydroxy butyrate concentrations were increased. Multiple regression analysis showed that at the end of the 13-week intervention, CLA did not affect %body weight regain (CLA LD 47.9+/-88.2%, CLA HD 27.4+/-29.8%, Placebo LD 32.0+/-42.8%, Placebo HD 22.5+/-37.9%). The regain of fat-free mass was increased by CLA (LD 6.2+/-3.9, HD 4.6+/-2.4%) compared to placebo (LD 2.8+/-3.2%, HD 3.4+/-3.6%), independent of %body weight regain and physical activity. As a consequence of an increased regain of fat-free mass by CLA, resting metabolic rate was increased by CLA (LD 12.0+/-11.4%, HD 13.7+/-14.4%) compared to placebo (LD 9.1+/-11.0%, HD 8.6+/-8.5%). Substrate oxidation and blood plasma parameters were not affected by CLA. CONCLUSION: In conclusion, the regain of fat-free mass was favorably, dose-independently affected by a 13-week consumption of 1.8 or 3.6 g CLA/day and consequently increased the resting metabolic rate. However, it did not result in improved body weight maintenance after weight loss.
Mb (myoglobin) plus H2O2 catalyses the oxidation of various substrates via a peroxidase-like activity. A Y103F (Tyr103-->Phe) variant of human Mb has been constructed to assess the effect of exchanging an electron-rich oxidizable amino acid on the peroxidase activity of human Mb. Steady-state analyses of reaction mixtures containing Y103F Mb, purified linoleic acid and H2O2 revealed a lower total yield of lipid oxidation products than mixtures containing the wild-type protein, consistent with the reported decrease in the rate constant for reaction of Y103F Mb with H2O2 [Witting, Mauk and Lay (2002) Biochemistry 41, 11495-11503]. Irrespective of the Mb employed, lipid oxidation yielded 9(R/S)-HODE [9(R,S)-hydroxy-10E,12Z-octadecadienoic acid] in preference to 13(R/S)-HODE [13(R,S)-hydroxy-9Z,11E-octadecadienoic acid], while 9- and 13-keto-octadecadienoic acid were formed in trace amounts. However, lipid oxidation by the Y103F variant of Mb proceeded with a lower V(max) value and an increased K(m) value relative to the wild-type control. Consistent with the increased K(m), the product distribution from reactions with Y103F Mb showed decreased selectivity compared with the wild-type protein, as judged by the decreased yield of 9(S)-relative to 9(R)-HODE. Together, these data verify that Tyr103 plays a significant role in substrate binding and orientation in the haem pocket of human Mb. Also, the midpoint potential for the Fe(III)/(II) one-electron reduction was shifted slightly, but significantly, to a higher potential, confirming the importance of Tyr103 to the hydrogen-bonding network involving residues that line the haem crevice of human Mb.
OBJECTIVE: We investigated the effects of specific conjugated linoleic acid (CLA) isomers on glucose metabolism and insulin resistance and on mRNA levels of genes important in glucose and lipid metabolism. METHODS: Sprague-Dawley rats were fed for 8 wk on a high-fat diet (45% kcal from fat) or one of three CLA-supplemented diets (1% CLA) containing differing isomers of CLA, including a mixture of CLAs (CLA mix), cis-9, trans-11-CLA (C9,T11-CLA), or trans-10, cis-12-CLA (T10,C12-CLA). RESULTS: Compared with the high-fat group, all the CLA groups had enhanced glucose tolerance. Insulin resistance index was significantly lower in the CLA-treated groups. No significant difference could be observed in the level of serum lipids between groups and in the activities of phosphoenolpyruvate carboxylase, glucose-6-phosphatase, and glucokinase. However, C9,T11-CLA and T10,C12-CLA significantly increased acyl coenzyme A oxidase mRNA in skeletal muscle. In addition, C9,T11-CLA increased hepatic acyl coenzyme A oxidase mRNA and skeletal muscle uncoupling protein-2 mRNA. The CLA mix showed intermediate effects on the levels of these genes. CONCLUSIONS: The addition of all types of CLA to Sprague-Dawley rats fed a high-fat diet can decrease insulin resistance. Possible mechanisms are increased fat oxidation and energy expenditure by increasing acyl coenzyme A oxidase and uncoupling protein-2 mRNA in the liver and/or skeletal muscle.
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