Conjugated linoleic acid alleviates glycolipid metabolic disorders by modulating intestinal microbiota and short-chain fatty acids in obese rats.

Although conjugated linoleic acid (CLA) has been shown to have anti-obesity properties, the effect and mechanism of CLA in alleviating glycolipid metabolism disorders remains unclear. In this work, it was observed that rats fed a high-fat diet (HFD) had lower body weight and body fat levels after 9 weeks of low-dose and high-dose CLA interventions. The results of blood biochemical indices showed that CLA significantly reduced the levels of total cholesterol, triglycerides, fasting blood glucose and insulin. Additionally, high-dose CLA could restore the intestinal microbiota composition, including increasing the relative abundances of short-chain fatty acid (SCFA)-producing microbiota, such as Dubosiella, Faecalibaculum and Bifidobacterium; decreasing the relative abundances of Enterococcus and Ruminococcus_2; and increasing the content of SCFAs in feces and serum. Further analysis showed that high-dose CLA could increase the expression levels of Insr, Irs-2, Akt and Glut4 in the liver tissue of HFD-induced obese rats. Consistently, high dose of CLA could reversibly improve the downregulation of INSR, AKT, PI3K and GLUT4 protein expression caused by HFD and reverse the decline in AKT phosphorylation levels. Correlation clustering analysis with a heatmap showed that the changes in specific microbiota induced by high-dose CLA were correlated with changes in obesity-related indices and gene expression. The molecular docking analysis showed that the molecular docking of SCFAs with the IRS-2, AKT and GLUT4 proteins had high linking activity. The results supported that CLA can alleviate glycolipid metabolic imbalances associated with obesity by altering the intestinal microbiota to induce the production of SCFAs and thereby activate the INSR/IRS-2/AKT/GLUT4 pathway. This study supports CLA may be preferentially used by the intestinal microbiota of the host to promote its health.

Multiple biological activities and biosynthesis mechanisms of specific conjugated linoleic acid isomers and analytical methods for prospective application.

Conjugated linoleic acid (CLA) is a collective term for the octadecadienoic acid isomers containing conjugated double bonds. This article reviewed CLA isomers from biological activities, biosynthesis mechanisms and analytical methods. The biological activities of CLA isomers in anti-obesity, cardiovascular protection, diabetes management and anti-cancer in vitro and in vivo were mainly reviewed. More attention has been paid to the production of the specific CLA isomer due to its biological activity. The biosynthesis methods of CLA isomers, such as dietary modification in ruminants and fermentation by microorganisms & enzymes, were systematically introduced. A rapid, accurate and economic analysis method will promote the research in both biological activities and biosynthesis mechanisms of CLA isomers. The merits of UV spectrometry, GC, HPLC, MS and CE used in the analysis of CLA isomers were also compared in detail. This paper aims to put into perspective the current status and future trends on CLA isomers.

Preparation of Conjugated Linoleic Acid by Ultrasound-assisted Nanonickel Catalyst Isomerization of Sunflower Oil.

A homemade nanonickel catalyst was made by the ultrasonic liquid-phase reduction method, characterized by X-ray diffraction, scanning electron microscopy and transmission electron microscopy, and applied to the isomerization reaction of high linoleic acid sunflower oil. Scanning electron microscopy (SEM), transmission electron microscopy (TEM) and particle size analysis showed that the homemade nickel particles were spherical, uniformly dispersed, less agglomerated, 20 to 75 nm in size, and nanoscale nickel powder. Compared with commercially available Raney nickel, the homemade nanonickel powder has a larger specific surface area, smaller pore size and higher catalytic activity. The X-ray diffraction spectrum of the homemade nanonickel powder had distinct diffraction peaks at its characteristic peaks which indicated that the powder was pure nickel. The nanometal nickel particles are fully dispersed in high oleic sunflower oil under the action of ultrasound. The results showed that it could effectively reduce the activation reaction time of nanonickel, and the conversion rate of conjugated linoleic acid could reach 86.24%. The process of activating the catalyst is omitted, the number of times of repeated uses of the nanonickel catalyst is increased, and the environmental pollution of the production is avoided. To obtain sunflower oil rich in CLA, it also provides a new idea for the preparation of conjugated linoleic acid.

Limited hydrolysis of glycosylated whey protein isolate ameliorates the oxidative and physical stabilities of conjugated linoleic acid oil-in-water emulsions.

Conjugated linoleic acid contains unsaturated fatty acids with multiple bioactivities, but it has poor oxidative and physical stabilities. Its emulsion was fabricated with glycosylated whey protein isolate and hydrolysates of glycosylated whey protein isolate to enhance its stability. An obvious decrease in peroxide value, thiobarbituric acid-reactive substances, particle size and creaming index of emulsion loaded by hydrolysates of glycosylated protein isolate with the increase of hydrolysis time. However, the absolute value of zeta-potential and interfacial adsorption rate of emulsion stabilized by hydrolysates of glycosylated whey protein isolate, were increased by 10.99 and 16.94% at hydrolysis time of 120 min, compared with emulsion loaded by glycosylated whey protein isolate. Thus, limited hydrolysis of glycosylated whey protein isolate as an effective method, remarkably improved the oxidative and physical stability of emulsion.

Laccase cross-linking of sonicated α-Lactalbumin improves physical and oxidative stability of CLA oil in water emulsion.

α-lactalbumin was modified by ultrasound (US, 20 kHz, 43 ± 3.4 W/cm-2) pre-treatments (0, 15, 30 and 60 min) and laccase cross-linking of sonicated α-lactalbumin was used to evaluate the physical and oxidative stability of conjugated linoleic acid (CLA) emulsions. The emulsions prepared with laccase cross-linking US-α-lactalbumin (α-lactalbumin treated with US pre-treatment) and US-α-lactalbumin were scrutinized for oxidative and physical stability at room temperature for two weeks of storage. Laccase cross-linking US-α-lactalbumin (Lac-US-α-lactalbumin) revealed improved physical stability in comparison with US-α-lactalbumin, specified by droplet size, structural morphology, adsorbed protein, emulsifying properties and creaming index. SDS-PAGE analysis showed that there was formation of polymers in Lac-US-α-lactalbumin emulsion. Surface hydrophobicity of Lac-US-α-lactalbumin was higher than that of US-α-lactalbumin, and gradually enhanced with the increase of ultrasound time. More importantly, the measurements of peroxide values and conjugated dienes were used to study the oxidative stability of the CLA emulsions. The Lac-US-α-lactalbumin emulsion proved to be reducing the synthesis of fatty acid hydroperoxides and less conjugated dienes compared to the native and US-α-lactalbumin emulsions. This study revealed that the combination of US pre-treatment and laccase cross-linking might be an effective technique for the modification of CLA emulsions.

Soy oil conjugated linoleic acid production with solar light photoisomerization.

The objectives were (1) to produce soy oil conjugated linoleic acid (CLA) triacylglycerides in large quantities with solar light photoisomerization, utilizing iodine as a photosensitizer, (2) to study the temperature variation in the photoisomerized oil during various hours of the day, and (3) to study the variations in solar light intensity during various hours of the day. A 0.5% iodine containing soy oil in glass box with a glass lid was photoisomerized, under natural solar light for 0, 11, and 27 days, and CLA isomers were determined with gas chromatography with flame ionization detector. After 27 days of solar light photoisomerization, the cis-9, trans-11 CLA; other cis, trans CLA; trans-10, cis-12 CLA; trans, trans CLA, and total CLA were found to be 0.62 ± 0.05%, 1.04 ± 0.09%, 0.54 ± 0.11%, 6.16 ± 0.68%, and 8.37 ± 0.90%, respectively. The concentration of CLA isomers between 0 and 11 days was significantly different (p < .05), and the concentration of CLA isomers between 0 and 27 days was also significantly different (p < .05). There is no significant difference (p > .05) in CLA concentration between 11 and 27 days treatment. The CLA was not found in control soy oil samples. The CLA isomers were measured with GDFID in 45 min instead of 120min. The temperature of the edible oil in glass boxes ranged from 26 °C (8 a.m.) to 56 °C (1 p.m.). The light intensity ranged from 4,146 lux (7 p.m.) to 95,490 lux (12 p.m.). Glass lid on the glass box affected light transmission to a small but statistically significant extent (p < .05). The CLA isomers could be energy efficiently and inexpensively produced in soy oil by solar light photoisomerization, at low temperature and without needing expensive reactor vessels or catalysts. PRACTICAL APPLICATION: CLA was produced effectively with the iodine sensitized solar light photoisomerization. CLA is produced in large quantities, inexpensively, for possible food additive applications. Produced CLA is in the form of stable triacylglycerides.

Enhancement of physicochemical and encapsulation stability of O1/W/O2 multiple nanoemulsions loaded with coenzyme Q10 or conjugated linoleic acid by incorporating polyphenolic extract.

Multiple O1/W/O2 nanoemulsions and O1/W nanoemulsions fortified with CLA or CoQ10 were produced using extra virgin olive or olive pomace oil and were also incorporated with polyphenols extracted from olive kernel to enhance their kinetic and chemical stability. They were prepared using a high-speed ultrasonic homogenizer. Specifically, nanoemulsions with 6 wt% lipid phase and 6 wt% non-ionic emulsifier (Tween 40) were produced and they demonstrated a droplet diameter >200 nm and high encapsulation stability during 30 days of storage at 4 °C or 25 °C. The incorporation of CLA or CoQ10 and polyphenolic compounds facilitated the homogenization of emulsions, reducing the droplet size and enhancing their chemical stability, and their bioactive retention values were >79%. O1/W/O2 nanoemulsions were produced using a mixture of non-ionic emulsifiers (Span 20 and Tween 40) and the O1/W enriched nanoemulsion as the dispersed phase. All multiple emulsions showed a bimodal droplet size distribution and Newtonian behavior while polyphenols facilitated their homogenization. Both vegetable oils resulted in samples with high kinetic and chemical stability; the bioactive retention values were found to be >80% at the end of 30 days of storage at 4 °C or 25 °C. Extra virgin olive oil resulted in more stable nanoemulsions in regards to kinetic and chemical stability at 4 °C, showing limited creaming and sedimentation boundary. Multiple nanoemulsions with the lowest initial droplet size presented the lowest droplet diameter growth and phase separation and the highest retention values. By comparing O1/W nanoemulsions and O1/W/O2 nanoemulsions, we noted that the reduction in the total phenolic content and antioxidant activity during storage was higher in the O1/W type. However, both delivery systems protected CLA and CoQ10 presenting high retention during storage. FTIR spectra before and after ultrasonic homogenization indicated that the sonication process did not significantly affect the lipid phase of O1/W/O2 nanoemulsions.

Decreased Metabolic Flexibility in Skeletal Muscle of Rat Fed with a High-Fat Diet Is Recovered by Individual CLA Isomer Supplementation via Converging Protective Mechanisms.

Energy balance, mitochondrial dysfunction, obesity, and insulin resistance are disrupted by metabolic inflexibility while therapeutic interventions are associated with improved glucose/lipid metabolism in skeletal muscle. Conjugated linoleic acid mixture (CLA) exhibited anti-obesity and anti-diabetic effects; however, the modulatory ability of its isomers (cis9, trans11, C9; trans10, cis12, C10) on the metabolic flexibility in skeletal muscle remains to be demonstrated. Metabolic inflexibility was induced in rat by four weeks of feeding with a high-fat diet (HFD). At the end of this period, the beneficial effects of C9 or C10 on body lipid content, energy expenditure, pro-inflammatory cytokines, glucose metabolism, and mitochondrial efficiency were examined. Moreover, oxidative stress markers, fatty acids, palmitoyletanolamide (PEA), and oleyletanolamide (OEA) contents along with peroxisome proliferator-activated receptors-alpha (PPARα), AKT, and adenosine monophosphate-activated protein kinase (AMPK) expression were evaluated in skeletal muscle to investigate the underlying biochemical mechanisms. The presented results indicate that C9 intake reduced mitochondrial efficiency and oxidative stress and increased PEA and OEA levels more efficiently than C10 while the anti-inflammatory activity of C10, and its regulatory efficacy on glucose homeostasis are associated with modulation of the PPARα/AMPK/pAKT signaling pathway. Our results support the idea that the dissimilar efficacy of C9 and C10 against the HFD-induced metabolic inflexibility may be consequential to their ability to activate different molecular pathways.

cis 9, trans 11, but not trans 10, cis 12 CLA isomer, impairs intestinal epithelial barrier function in IPEC-J2 cells and mice through activation of GPR120-[Ca2+]i and the MLCK signaling pathway.

This study aimed to investigate the effects of conjugated linoleic acid (CLA) on intestinal epithelial barrier function and explore the underlying mechanisms. IPEC-J2 cells and mice were treated with different CLA isomers. The intestinal epithelial barrier function determined by transepithelial electrical resistance (TEER), the expression of tight junction proteins, and the involvement of G-protein coupled receptor 120 (GPR120), intracellular calcium ([Ca2+]i) and myosin light chain kinase (MLCK) were assessed. In vitro, c9, t11-CLA, but not t10, c12-CLA isomer, impaired epithelial barrier function in IPEC-J2 by downregulating the expression of tight junction proteins. Meanwhile, c9, t11-CLA isomer enhanced GPR120 expression, while knockdown of GPR120 eliminated the impaired epithelial barrier function induced by c9, t11-CLA isomer. In addition, c9, t11-CLA isomer increased [Ca2+]i and activated the MLCK signaling pathway in a GPR120-dependent manner. However, chelation of [Ca2+]i reversed c9, t11-CLA isomer-induced MLCK activation and the epithelial barrier function impairment of IPEC-J2. Furthermore, inhibition of MLCK totally abolished the impairment of epithelial barrier function induced by c9, t11-CLA. In vivo, dietary supplementation of c9, t11-CLA rather than t10, c12-CLA isomer decreased the expression of intestinal tight junction proteins and GPR120, increased intestinal permeability, and activated the MLCK signaling pathway in mice. Taken together, our findings showed that c9, t11-CLA, but not t10, c12-CLA isomer, impaired intestinal epithelial barrier function in IPEC-J2 cells and mice through activation of GPR120-[Ca2+]i and the MLCK signaling pathway. These data provided new insight into the regulation of the intestinal epithelial barrier by different CLA isomers and more references for CLA application in humans and animals.

Antiproliferation Activity and Mechanism of c9, t11, c15-CLNA and t9, t11, c15-CLNA from Lactobacillus plantarum ZS2058 on Colon Cancer Cells.

Conjugated linolenic acid (CLNA) is a type of ω-3 fatty acid which has been proven to have a series of benefits. However, there is no study about the function of Lactobacillus-derived CLNA isomer. Lactobacillus plantarum ZS2058 has been proven to manifest comprehensive functions and can produce CLNA. To investigate the specific functions of CLNA produced by this probiotic bacterium, two different conjugated α-linolenic acid (CLNA) isomers were successfully isolated. These isoforms, CLNA1 (c9, t11, c15-CLNA, purity 97.48%) and CLNA2 (c9, t11, t15-CLNA, purity 99.00%), both showed the ability to inhibit the growth of three types of colon cancer cells in a time- and concentration-dependent manner. In addition, the expression of MDA in Caco-2 cells was increased by CLNA1 or CLNA2, which indicated that lipid peroxidation was related to the antiproliferation activity of CLNAs. An examination of the key protein of pyroptosis showed that CLNA1 induced the cleavage of caspase-1 and gasdermin-D, while CLNA2 induced the cleavage of caspase-4, 5 and gasdermin-D. The addition of relative inhibitors could alleviate the pyroptosis by CLNAs. CLNA1 and CLNA2 showed no effect on caspase-3, 7, 9 and PARP-1, which were key proteins associated with apoptosis. No sub-diploid apoptotic peak appeared in the result of PI single staining test. In conclusion, CLNA1 activated caspase-1 and induced Caco-2 cell pyroptosis, whereas CLNA2 induced pyroptosis through the caspase-4/5-mediated pathway. The inhibition of Caco-2 cells by the two isomers was not related to apoptosis. This is the first study on the function of Lactobacillus-derived CLNA isomer. The inhibition pathway of Lactobacillus-derived CLNA isomer on colon cancer cells were proved.

Conjugated Linoleic Acid Isomers Affect Profile of Lipid Compounds and Intensity of Their Oxidation in Heart of Rats with Chemically-Induced Mammary Tumors-Preliminary Study.

Breast cancer and cardiovascular diseases (CVD) have shared risk factors and mechanisms of pathogenicity, as proven by increased cardiac risk in breast cancer patients receiving anticancerogenic therapies and in cancer survivors. A growing mammary tumor may cause heart injury in cancer patients who have not yet been treated. This study aimed to evaluate the effect of conjugated linoleic acid (CLA) supplementation of female rats with 7,12-dimethylbenz(a)anthracene (DMBA)-induced cancerogenesis on fatty acids (FAs), conjugated FAs (CFAs), malondialdehyde (MDA), cholesterol and oxysterols content in cardiac tissue. FAs, cholesterol and oxysterols contents were determined by gas chromatography coupled with mass spectrometry, while the contents of CFAs and MDA were determined by high performance liquid chromatography with photodiode detection. Our results indicate that both CLA supplementation and the presence of tumors influence the lipid biomarkers of CVD. A significant interaction of both experimental factors was observed in the content of polyunsaturated FAs (PUFAs), n-6 PUFAs and CFAs. CLA supplementation significantly inhibited PUFA oxidation, as evidenced by the lower content of MDA in rats' hearts, while the cancerous process intensified the oxidation of cholesterol, as confirmed by the elevated levels of 7-ketocholesterol in DMBA-treated rats. These results may significantly expand knowledge about CLA properties in terms of the prevention of co-existing non-communicable diseases.

Cis-9, Trans-11 Conjugated Linoleic Acid Reduces Phosphoenolpyruvate Carboxykinase Expression and Hepatic Glucose Production in HepG2 Cells.

Dysregulated hepatic gluconeogenesis is a hallmark of insulin resistance and type 2 diabetes mellitus (T2DM). Although existing drugs have been proven to improve gluconeogenesis, achieving this objective with functional food is of interest, especially using conjugated linoleic acid (CLA) found in dairy products. Both cis-9, trans-11 (c9,t11) and trans-10, cis-12 (t10,c12) isomers of CLA were tested in human (HepG2) and rat (H4IIE) hepatocytes for their potential effects on gluconeogenesis. The hepatocytes exposed for 24 h with 20 µM of c9,t11-CLA had attenuated the gluconeogenesis in both HepG2 and H4IIE by 62.5% and 80.1%, respectively. In contrast, t10,c12-CLA had no effect. Of note, in HepG2 cells, the exposure of c9,t11-CLA decreased the transcription of gluconeogenic enzymes, cytosolic phosphoenolpyruvate carboxykinase (PCK1) by 87.7%, and glucose-6-phosphatase catalytic subunit (G6PC) by 38.0%, while t10,c12-CLA increased the expression of G6PC, suggesting the isomer-specific effects of CLA on hepatic glucose production. In HepG2, the peroxisome proliferator-activated receptor (PPAR) agonist, rosiglitazone, reduced the glucose production by 72.9%. However, co-administration of c9,t11-CLA and rosiglitazone neither exacerbated nor attenuated the efficacy of rosiglitazone to inhibit glucose production; meanwhile, t10,c12-CLA abrogated the efficacy of rosiglitazone. Paradoxically, PPARγ antagonist GW 9662 also led to 70.2% reduction of glucose production and near undetectable PCK1 expression by abrogating CLA actions. Together, while the precise mechanisms by which CLA isomers modulate hepatic gluconeogenesis directly or via PPAR warrant further investigation, our findings establish that c9,t11-CLA suppresses gluconeogenesis by decreasing PEPCK on hepatocytes.

Conjugated Linolenic Acids: Implication in Cancer.

Conjugated fatty acids (CFAs) including both conjugated linoleic acids (CLAs) and conjugated linolenic acids (CLNAs) have various health promoting effects. These beneficial effects are comprised by their antioxidant, antiatherogenecity, anticarcinogenic activities, etc. Several reports indicate that CLNAs such as eleostearic acid, punicic acid, jacaric acid, and calendic acid possess anticancer properties. These CLNAs are produced and accumulated in seeds of certain commonly available plants. This review discusses their role in chemoprevention of cancer. Using in vitro as well as in vivo models of cancer, bioactivities of these CLNAs have been explored in detail. CLNAs have been shown to have potent anticancer activity as compared to the CLAs. Although the molecular basis of these effects has been summarized here, more detailed studies are needed to explore the underlying mechanisms. Further clinical trials are obligatory for assessing the safety and efficacy of CLNAs as an anticancer agent.

Effect of frying and storage on oxidative quality of conjugated linoleic-acid-rich soybean oil produced by photoisomerization using plantain as a model system.

BACKGROUND: Conjugated linoleic acid (CLA) is known to have beneficial properties to health. Naturally, in foods it is found in very low concentrations, and so these beneficial properties cannot be obtained. This study investigated the enrichment of soybean oil by photoisomerization, as well as assessing its oxidative stability during the frying process using plantain slices as a model system and after a storage period of 20 days at 60 °C. RESULTS: The oxidative stability of soybean oil enriched with CLA by photoirradiation was measured based on the peroxide, p-anisidine, and Totox values, as well as by the polyphenol content, tocopherol content and DPPH· scavenging capacity. The results obtained showed that a substantial amount of CLA was obtained by photoirradiation (31.73%). The oxidative stability values of the oil enriched with CLA showed good stability during a frying cycle; however, this stability decreased when it was stored and during the final frying cycles. CONCLUSIONS: The results obtained indicated that photoirradiation is a good technique for obtaining oils enriched with CLA, and in this way CLA can be incorporated into foods; however, it is necessary to add antioxidants to improve their stability. © 2019 Society of Chemical Industry.

Regulation of Stearoyl-Coenzyme A Desaturase 1 by trans-10, cis-12 Conjugated Linoleic Acid via SREBP1 in Primary Goat Mammary Epithelial Cells.

trans-10, cis-12 Conjugated linoleic acid (t10c12-CLA) is a biohydrogenation intermediate in the rumen that inhibits mammary fatty acid de novo synthesis in lactating dairy goats. However, the underlying molecular pathways in milk-lipid metabolism affected by t10c12-CLA are not completely understood. The present study investigated the lipid-regulation mechanisms in goat mammary epithelial cells (GMECs) in response to t10c12-CLA. Gene-expression analysis indicated sterol-regulatory-element-binding transcription factor1 ( SREBF1) and its putative target gene stearoyl-CoA desaturase ( SCD1) were down-regulated (fold changes of 0.33 ± 0.04, P < 0.05, and 0.19 ± 0.01, P < 0.01, respectively). Concentrations of cellular palmitoleic acid (C16:1) and oleic acid (C18:1) were decreased (1.12 ± 0.05 vs 1.69 ± 0.11% and 15.70 ± 0.44 vs 24.97 ± 0.82%, respectively, P < 0.01), whereas those of linoleic acid (C18:2) were increased (5.00 ± 0.14 vs 3.81 ± 0.25%, P < 0.05); the desaturation indices of C16 and C18 were decreased in response to t10c12-CLA treatment (6.90 ± 0.05 vs 8.00 ± 0.30% and 61.41 ± 0.65 vs 67.73 ± 1.33%, respectively, P < 0.05). A luciferase-activity assay indicated that deletion of the sterol-response-element (SRE) site and the nuclear-factor (NF-Y) site in the SCD1-promoter region (-511/+65 bp) suppressed the regulatory effect of t10c12-CLA. Overexpression of SREBF1 partly counteracted the inhibitory effect of t10c12-CLA on de novo fatty acid synthesis. Overall, t10c12-CLA causes an inhibition of fatty acid synthesis and desaturation and regulates SCD1 expression by affecting the binding of SREBP1 protein to the SRE and NF-Y sites.

How to safeguard an appropriate "all trans retinoic acid" concentration to keep cell division on track: Exploring therapeutic hotspots from metabolomics.

In this letter to editor, I hypothesize a potential affinity of retinol saturase (RetSat) enzyme towards a conjugated trienoic fatty acid; alpha-eleostearic acid (α-ESA) and subsequent hindrance of the action on its usual substrate; all trans retinol. Hence, RetSat is speculated to be involved in a rapid unusual conversion of α-ESA to conjugated linoleic acid (CLA), giving a less priority to its usual substrate all trans retinol, which would subsequently be converted into "all trans retinoic acid" (atRA). Otherwise, all trans retinol is converted by RetSat into all-trans-13,14-dihydroretinol and eventually forms all-trans-13,14-dihydroretinoic acid, but not the atRA. The atRA controls differentiation, proliferation and apoptosis of cells and it's deficiencies end up as neoplasms. Thus, here it is emphasized that safeguarding atRA would help controlling cell division and growth in a favourable manner. Hence, inhibition of RetSat could be a hot target to control unwarranted cell growths within the body. This hypothesis could be easily tested in a RetSat ablated (RetSat -/-) animal model or using antagonists on RetSat activity or α-ESA.

PPARs are mediators of anti-cancer properties of superparamagnetic iron oxide nanoparticles (SPIONs) functionalized with conjugated linoleic acid.

Breast cancer chemotherapy can cause side effects due to nonspecific drug delivery, low solubility and fast metabolism of drugs used in conventional therapy. Moreover, the therapeutic effect of the drugs is often reduced by the strengthening of chemoresistance, which occurs via a variety of mechanisms. Different strategies have been developed to reduce multidrug resistance (MDR)-associated gene expressions including the use of surfactants and polymers. In this study superparamagnetic iron oxide nanoparticles (SPIONs) functionalized with conjugated linoleic acid (CLA) reduced the number and viability of cells in comparison with both untreated cells or cells treated with SPIONs alone. This cytostatic effect correlated with the increase of peroxisome proliferator-activated receptors γ (PPARγ). The necrotic cell death induced, as a consequence, an inflammatory process, as evidenced by the decrease of the anti-inflammatory PPARα and increase of pro-inflammatory TNFα and IL-1ß. PPARs were examined because CLA is one of their natural ligands. The antitumor effect observed was accompanied by a down-regulation of p-glycoprotein (P-gp), which was the first important discovered efflux transporter belonging to MDR, and of ALDH3A1, an enzyme able to metabolize some drugs, reducing their effects. The down-regulation of P-gp correlated with the increase of cytokines. The ALDH3A1 decrease correlated with the increase of PPARγ. Based on these results, PPARs are molecular mediators of anti-cancer effect of SPIONs functionalized with CLA, being changes in these nuclear receptors correlated with induction of cytotoxicity and inflammation, and decreased ability of cancer cells in blocking anti-cancer drug effect.

Fatty Acids of CLA-enriched Egg Yolks Can Induce Mitochondrial Pathway of Apoptosis in MCF-7 Breast Cancer Cells.

BACKGROUND: Fatty acids from conjugated linoleic acid (CLA)-enriched egg yolk suppressed the viability of the MCF-7 cancer line more effectively than non-enriched egg yolk. Herein we aimed to determine the molecular mechanisms by analysing the expression and activation of proteins involved in cellular stress and apoptosis signaling. MATERIALS AND METHODS: Forty-eight Isa Brown laying hens (26-week-old) were fed a fortified (0.75% CLA) or a control diet (0% CLA) for 4 months. Collected eggs were used to obtain CLA-enriched (EFA-CLA) or non-enriched (EFA) fatty acid extracts for the treatment of the MCF-7 cancer cell line. Protein levels were analysed by PathScan® Stress and Apoptosis Signalling Antibody Array and western blot method. RESULTS: Treatment with EFA-CLA led to activation of caspase signalling as main effector of apoptosis. It also increased levels of pro-apoptotic B-cell lymphoma 2 family proteins as well as promoted the release of cytochrome c, second mitochondria-derived activator of caspase and mitochondrial serine protease from mitochondria to the cytoplasm. EFA-CLA increased levels of tumour protein 53 and mothers against decapentaplegic homolog 2 tumour suppressors, and activated p38 mitogen-activated protein kinases and stress-activated protein kinase/c-Jun NH2-terminal kinase proteins. Finally, treatment down-regulated anti-apoptotic extracellular signal-regulated protein kinases 1 and 2, RAC-alpha serine/threonine-protein kinase, heat-shock protein 27, inhibitor of nuclear factor κß, transforming growth factor beta-activated kinase 1 and survivin proteins. CONCLUSION: Taken together, our results suggest that activation of the mitochondrial apoptotic pathway may be a potential mechanism of EFA-CLA action.

Nano-in-Micro Delivery System Prepared by Co-Axial Air Flow for Oral Delivery of Conjugated Linoleic Acid.

The preparation of a nano-in-micro delivery system (NiMDS) under mild conditions without using toxic organic solvents and expensive equipment still faces challenges. In this study, we introduced the co-axial air flow method to prepare NiMDS for the oral delivery of conjugated linoleic acid (CLA). The chitosan nanoparticles were prepared using the stearic-acid-modified chitosan through self-aggregation. Then, the chitosan nanoparticles were incorporated into alginate microparticles by the co-axial air flow method. The obtained chitosan nanoparticles and NiMDS were spherical in shape with the average sizes of 221⁻243 nm and 130⁻160 µm, respectively. Compared with alginate microparticles, the hybrid particles were of fewer fragments, were bigger in size, had a higher mechanical strength, and showed a controlled release in the phosphate buffer solution (pH 1.2 or 7.4). The release kinetics study showed that encapsulating the chitosan nanoparticles into the alginate microparticles inhibited the dissolution of alginate microparticles at the initial stage. These results revealed the potential of NiMDS as an ideal oral carrier for the sustained release of CLA in the gastrointestinal environment.

Bacterial conjugated linoleic acid production and their applications.

Conjugated linoleic acid (CLA) has been shown to exert various potential physiological properties including anti-carcinogenic, anti-obesity, anti-cardiovascular and anti-diabetic activities, and consequently has been considered as a promising food supplement. Bacterial biosynthesis of CLA is an attractive approach for commercial production due to its high isomer-selectivity and convenient purification process. Many bacterial species have been reported to convert free linoleic acid (LA) to CLA, hitherto only the precise CLA-producing mechanisms in Propionibacterium acnes and Lactobacillus plantarum have been illustrated completely, prompting the development of recombinant technology used in CLA production. The purpose of the article is to review the bacterial CLA producers as well as the recent progress on describing the mechanism of microbial CLA-production. Furthermore, the advances and potential in the heterologous expression of CLA genetic determinants will be presented.