Printer Friendly

Effect of Supplementation with n-3 Fatty Acids Extracted from Microalgae on Inflammation Biomarkers from Two Different Strains of Mice.

1. Introduction

Diabetes mellitus is a multifactorial chronic noncommunicable disease, characterized by states of hyperglycemia resulting from defects in insulin secretion, its action, or both [1]. In Mexico, according to the National Health and Nutrition Survey HalfWay 2016 (ENSANUT MC 2016 for its acronym in Spanish), 9.4% of the adult population has been diagnosed with diabetes [2]. It also represents the leading cause of negative health outcomes such as heart failure, blindness, kidney failure, amputations, and premature death [3]. The main cause of health complications in diabetes is chronic hyperglycemia, which is associated with changes in immunomodulation and inflammation [4].

The use of n-3 polyunsaturated fatty acids as a strategy to minimize damage caused by hyperglycemia has been deeply studied [5]. Its biological effects include benefits on the metabolism of lipoproteins [6], platelet, and endothelial and vascular function [7], as well as antioxidant and antiinflammatory impact [8]. Evidence suggests that n-3 inhibits the proliferation of T lymphocytes in murine models and in humans [9, 10] and inhibits degranulation of cytotoxic T lymphocytes [11]. Thus, it suggests that polyunsaturated fatty acids have potentially immunosuppressive properties. Moreover, the supplementation with EPA (eicosapentaenoic acid) for 12 weeks has been shown to modify the fatty acids composition of the phospholipids of plasma, platelets, neutrophils, monocytes, and T and B lymphocytes [12].

Within their anti- and proinflammatory effects, it has been shown that, in cell cultures, EPA and DHA (docosahexaenoic acid) have high anti-inflammatory and immunosuppressive effects [13-15]. Same findings have been shown on animal studies supplemented with fish oil [16-18]. It has been proven that EPA and DHA supplementation decreases proinflammatory cytokines such as tumor necrosis factor Alpha (TNF-[alpha]), interleukin-6 (IL-6), monocyte-1 chemoattractant protein (MCP-1), and plasminogen activator-1 (PAI1) inhibitor [19].

The main dietary sources of EPA and DHA fatty acids are fish, shellfish, and marine oils [20]. However, some disadvantages of the use of these sources are undesirable nutritional and organoleptic effects, such as oxidation (due to their high polyunsaturation) and the characteristic odor of the product [21]. Another disadvantage of the use of marine oils is the risk of contamination with heavy metals and pesticides, the solution of which may be oil refining, but this process involves a higher cost of production [22]. In addition, in recent years, marine sources have been diminished because fish catch has exceeded the maximum sustainable levels [23].

Microalgae are an evolutionarily microscopic diverse eukaryotic group of unicellular and predominantly aquatic photosynthetic organisms that have recently been studied for their potential to produce compounds of high biological value and beneficial for health such as carotenoids, polyunsaturated fatty acids, and very long chain polyunsaturated fatty acids [24]. They are the primary natural producers of EPA and DHA, because they have the biosynthetic machinery to sequentially alternate between desaturation and elongation in their carbon chains [25]. Because microalgae are at the beginning of the food chain, fish are the main consumers of these fatty acids, which is why they are incorporated into the lipids of membranes and accumulated in the fats and meat of many marine species [24]. However, there are fewer studies describing their anti-inflammatory effects as those already described for fatty acids of animal origin [26-28].

For all the above and the growing study of the composition and properties of microalgae, as well as the possibility of cultivating them in artificial form, interest has aroused in the study of these microorganisms as a renewable source of n-3 fatty acids. The aim of this study was to analyze the effect of the consumption of n-3 fatty acids extracted from microalgae (Chlorophyceae and Eustigmatophyceae families) either provided as a supplement or incorporated to diet, on the inflammatory markers from two different strains of mice: db/db mice as a model of obesity and diabetes mellitus in which an inflammation state is expected and CD1 mice as a model of optimal state of health without inflammation.

2. Methods

2.1. Animals and Study Groups. The present experimental, prospective, controlled, and randomized study was conducted with sixty 8-week-old male mice from two different strains: 30 db/db mice (BKS.Cg + Leprdb + LeprdbOlaHsd Harlan[R]) and 30 CD1 mice (Crl: cD1 (ICR) belonging to the Faculty of Medicine from the Autonomous University of Mexico State). For each strain, five study groups were formed (n = 6): (1) a baseline (BL) group to obtain baseline values; (2) a Rodent Chow (RC) group; (3) a RC + lyophilized microalgae n-3 fatty acids (LY) group; (4) a RC + saturated fatty acid (SAT) group; (5) modified diet (MD) supplemented with microalgae n-3 fatty acids group. Supplementation was administered from 8th to 16th week of life (Figure 1). The animals were housed in acrylic cages of 19 x 29 x 13 cm, with light/dark cycles of 12/12 h with controlled temperature at 21 [+ or -] 1[degrees]C. Groups 2, 3, and 4 were fed a standard normal diet (Rodent Laboratory Chow 5001 from Purina [3.02kcal/g]) and water ad libitum. Water consumption (mL/week) and food (g/week) were recorded weekly. Animal care and experimental procedures in rodents were carried out in accordance with the rules of the Internal Regulations for the Use of Laboratory Animals and the Committee of Ethics in Research of the UAEMex, as well as the guidelines of the Ministry of Health and Agriculture of Mexico for the Production and Care of Laboratory Animals (NOM-062-ZOO-1999), Mexico City, Mexico. This protocol was approved by the Ethical Research Committee from the Faculty of Medicine of the UAEMex.

2.2. Obtaining of n-3 Fatty Acids (EPA and DHA) from Microalgae. The microalgae used in this project were native and collected and isolated by BIOMEX SA. de CV. The strains used were from Chlorophyceae and Eustigmatophyceae families which have a high content of EPA and DHA. The use of these microalgae for such purpose is of recent interest. The process for obtaining the EPA and DHA includes the cultivation of microalgae, separation of biomass, extraction of total lipids, and finally chromatographic procedures for EPA and DHA content determination (25.7% EPA + DHA). EPA and DHA were provided as free fatty acids form.

2.3. Supplementation. (a) LY group was fed with Rodent Chow and supplemented with lyophilized powder containing EPA + DHA obtained from microalgae. The supplemental dose was 1mg/g of mouse weight, reconstituted in 100 [micro]l of distilled water, and administered with micropipette by direct oral deposition every day at 8:00 am.

(b) SAT group was fed with Rodent Chow and supplemented with coconut oil. The daily dose of coconut oil was 1 mg/g of mouse weight administered with micropipette by oral deposition at 8:00 am.

(c) MD group was fed with a Rodent Chow added with microalgae EPA + DHA for a total content of 2.0% n-3 fatty acid which means 10x the original content; Chow was administered ad libitum (Table 1).

2.4. Determination of Body Mass Index (BMI) and Blood Glucose Concentration. The BMI and blood glucose concentrations of animals were determined at the 8th and 16th week of life. The formula used for BMI determination was BMI = [weight (g)/length (cm)2 * 100]. Weight was determined using a mouse Triple Beam 700/800 series Ohaus[R] brand weighing scale and length was determined by measuring the animal from nose to anus. Blood glucose was determined with a Bayer Contour TS glucometer through tail puncture.

2.5. Collection of Biological Samples. The BL groups were sacrificed at the 8th week of life and the rest of the groups were sacrificed at the 16th week of life. Animals were anesthetized by ether camera, bled by direct cardiac puncture (using a heparinized syringe, obtaining 1mL of blood), and then sacrificed by cervical dislocation. 500 [micro]lof the collected blood was used for lymphocyte isolation using Ficoll-Hypaque Plus (GE Healthcare Bio-Sciences AB, Sweden); lymphocytes were stored with a PBS (phosphate-buffered saline) solution to obtain a final volume of 1 ml for further flow cytometry.

2.6. Flow Cytometry Assays. Cell suspensions of peripheral blood mononuclear cell (PBMC) were adjusted to 1 x 106 cells/mL in PBS for the cytofluorometric analysis with brief modifications [29]. (i) Surface phenotype of T cells was detected by using fluorescent labeled monoclonal antibodies: anti-CD3 FITC (Cat. number 553063), anti-CD8[alpha] PE (Cat. number 553035), and anti-CD4 PerCP (Cat. number 553052) (all from BD Biosciences). Cells were incubated for 30 min at room temperature. Finally, the cells were then washed with PBS and fixed in 1% paraformaldehyde. (ii) For the detection of intracellular cytokine production, lymphocytes were stimulated with a mixture containing phorbol myristate acetate, ionomycin, and Brefeldin A (Leucocyte Activation Cocktail Kit, BD Pharmingen) and incubated for 4h at 37[degrees] C and 5% CO2. Then, antibodies to cell surface markers, anti-CD4 PerCP, were added and incubated as before. For intracellular staining of CD4+ T cells, fixation and permeabilization were performed using Cytofix/Cytoperm Kits (BD Pharmingen) according to the manufacturer's instructions. These cells were incubated with anti-IL-4 PE (Cat. number 554435), anti-IL5 PE (Cat. number 554395), anti-IL-6 APC (Cat. number 561367), anti-IL-10 FITC (Cat. number 554466), anti-IL-17A FITC (BioLegend Cat. number 506907), anti-IFN-[gamma] FITC (Cat. number 554411), and anti-TNF-[alpha] PE antibodies (Cat. number 554419). For all samples, the expression of CD69 was measured as an activation control. The fluorescent signal intensity was recorded and analyzed by FACS Aria Flow Cytometer (Becton Dickinson). Events were collected from the lymphocyte gate on the FSC/SSC dot plot. 20,000 gated events were acquired from each sample using the CellQuest research software (Becton Dickinson). Data was analyzed using Summit software v4.3 (Dako, Colorado Inc.). Data from six mice per group are reported as the mean [+ or -] standard deviation (SD).

2.7. Statistical Analysis. One-way ANOVA was performed for comparison between groups from each strain (BL, RC, LY, SAT, and MD); Bonferroni post hoc test was applied. Differences were considered significant at p < 0.05. Software used to run statistical analysis was SPSS v.23 for Windows.

3. Results

3.1. BMI Was Higher in the MD Group for Diabetic Mice and Blood Glucose Was Higher in All the db/db Groups. In the diabetic db/db mice, the MD group showed a significantly higher BMI than the BL group; blood glucose concentrations were significantly higher in all groups compared to the BL group; the food intake was significantly lower in the MD group and higher in the LY group and finally the water consumption was significantly higher in the MD group, all of this compared with the BL group (Bonferroni post hoc, p < 0.001). In the healthy mice (CD1), there were no significant differences in BMI and blood glucose. On the other hand, the consumption of food was significantly higher in the SAT group and the water consumption was significantly higher in the LY and SAT groups, all this compared with the BL group (Bonferroni post hoc, p < 0.001) (Table 2).

3.2. Microalgae Fatty Acids Modified Lymphocyte Populations in db/db Mice by Lowering CD3+ and CD8+ Populations and in CD1 Mice by Lowering CD3+. In the db/db strain, the percentage of CD3+ lymphocytes was significantly higher in all the groups when compared to the BL group (Bonferroni post hoc, p < 0.001). Regarding CD4+ lymphocytes, the MD group showed a significantly lower percentage and the SAT group a higher percentage (Bonferroni post hoc, p < 0.001). Finally, the CD8 + lymphocytes have a higher percentage in the MD, LY, and SAT groups (Bonferroni post hoc, p < 0.001).

For CD1 strain mice, the percentage of CD3+ lymphocytes was lower in the MD, LY, and SAT groups and higher in the RC group, all compared with the BL group (Bonferroni post hoc, p < 0.001). Regarding CD4 + lymphocytes, the percentage was significant in the MD and SAT groups (Bonferroni post hoc, p < 0.001). Finally, the percentage of CD8+ lymphocytes was significantly higher in the RC and LY groups, but lower in the MD and SAT groups, all compared with the BL group (Bonferroni post hoc, p < 0.001) (Table 3).

3.3. Supplementation with Microalgae Fatty Acids Increases IL17A, IL-12, IL-4, IL-6, IL-10, and TGF-[beta] but Decreases IFN-[gamma], TNF-[alpha], and IL-5 in Diabetic Mice. In the diabetic mice, the behavior of Th1 type cytokines was the same for all study groups; the proportion of TCD4+ cells producing IFN-[gamma] and TNF-a was significantly lower in all groups compared to the BL (Bonferroni post hoc, p < 0.001). On the contrary, the percentage of TCD4+ cells producing IL-12 and IL-17A was significantly lower in all groups (Bonferroni post hoc, p < 0.001). As for the Th2 type cytokines, the percentage of TCD4+ cells producing IL-4 was higher in the RC, MD, and SAT groups and, on the other hand, it was lower in the LY group. The percentage of TCD4+ cells producing IL-5 was significantly lower in all groups compared to the baseline group (Bonferroni post hoc, p < 0.001). For all groups, the percentage of TCD4+ cells producing IL-6 and IL-10 was significantly lower compared to the initial group (Bonferroni post hoc, p < 0.001). Finally, the percentage of TCD4+ cells pro ducing TGF-[beta] was significantly lower in the RC group and higher in the MD, LY, and SAT groups (Table 4).

3.4. Supplementation with Microalgae Fatty Acids Lyophilized Increases IL-17A but Decreases IFN-[gamma], TNF-a, IL-12, IL-4, and IL-6 in Healthy Mice. In CD1 mice, for Th1 type cytokines, the percentage of TCD4+ cells producing IFN-[gamma] and TNF[alpha] was significantly lower in the RC and LY groups and significantly higher in the SAT group (Bonferroni post hoc, p < 0.001). The percentage of TCD4+ cells producing IL-12 was significantly lower in the RC, MD, and LY groups and higher in the SAT group, all compared with the BL group (Bonferroni post hoc, p < 0.001); finally, the percentage of TCD4+ producing IL-17A was significantly lower in all groups compared to the BL group (Bonferroni post hoc, p < 0.001). Regarding the Th2 type cytokines, the SAT group showed a significantly higher percentage of TCD4+ cells producing IL-4, IL-5, IL-6, and TGF-[beta] compared to the BL group (Bonferroni post hoc, p < 0.001), and the MD group showed similar behavior for IL-5 and TGF-[micro]. On the other hand, the percentage of TCD4+ cells producing IL-4 and IL-6 was significantly lower in the LY groups (Bonferroni post hoc, p < 0.001) (Table 5).

4. Discussion

The results provided in this study show evidence that supplementation with n-3 fatty acids obtained from microalgae improves the inflammatory profile in general by reducing the secretion of many cytokines. Therefore, these results suggest that microalgae extracts may be considered as an anti-inflammatory strategy against different diseases. These findings are summarized in Figure 2.

The BMI was significantly higher in the MD group from the diabetic mice. These results are different from those reported by Zhuang in which C57B1/6 mice were supplemented with fish oil and did not show changes in BMI [30]. There were no significant differences in plasma glucose attributable to treatment; no studies were found to match our findings; however, a study with C57B/6 mice supplemented with EPA suggests a protective effect of n-3 fatty acids on glucose metabolism [19]. Further studies on the effect of microalgae fatty acids on glucose metabolism are needed.

In this study, food consumption was lower in the MD groups for the diabetic mice. Other studies with C57Bl/6 mice fed with normal chow enriched with EPA and DHA extracted from fish oil showed no changes in food consumption [31,32]. However, a study from Diaz- Resendiz explains that mice regulate food intake according to the composition of the food or the presence of an extra source of energy [33].

The percentage of total T lymphocytes was lower in all study groups from both strains. In contrast to this study, Marano et al. [34] suggest that consumption of n-3 fatty acids increases CD3+ lymphocyte populations including CD4+. In agreement with our findings, several supplementation studies report that there were no changes in lymphocyte populations [35,36].

On the other hand, in both strains the SAT groups showed a significantly lower percentage of CD8+ cells. The CD4+ populations in the SAT groups increased significantly compared to their BL group. These results are consistent with those of Baccan et al. [37] who showed that consumption of high-fat diets significantly increases lymphocyte populations.

The db/db strain is characterized by a chronic inflammatory state such as diabetes disease, which causes the pro- and anti-inflammatory cytokines to be in higher concentrations compared to the CD1 strain; however, although the strains are very different between them, the decrease in cytokine concentrations occurred in a similar way.

In this study, supplementation with n-3 fatty acids extracted from microalgae significantly decreased the percentage of TCD4+ cells producing IFN-[gamma] and TNF-[alpha]. These cytokines play different roles in inflammatory states such as in diabetes; IFN-[gamma] directs the differentiation of CD4+ lymphocytes into helper lymphocytes type 1 (Th1); it also intervenes in the activation of macrophages and induces a greater secretion of IL-12 [38]. However, in the diabetic mice, microalgae fatty acids were shown to increase the percentage of TCD4+ cells producing IL-12. The main functions of IL-12 are the activation of Th1 lymphocytes and to stimulate the production of IFN-[gamma] [39]. On the other hand, TNF-[alpha] is a cytokine involved in the acute and chronic phase as well as in the activation of the production of certain anti-inflammatory cytokines such as IL-4, IL-5, and IL-6 as a form of self-regulation of the inflammatory state [40].

A study by Vigerust et al. made in transgenic TNF-[alpha] C57B/6 mice that were fed with diets enriched with either fish oil or krill oil showed no modification in this cytokine [40]. Similarly, in a study on Wistar rats supplemented with fish or soybean oil, no significant differences were found in the concentrations of IFN-[gamma] and TNF-[alpha] [41]. Although Xavier et al. [41] showed there are no changes in TNF-[alpha] after fish oil supplementation, there are also many studies that report a significant decrease in this cytokine [42-44]. A study made by Sierra et al. [45] reports that, in Balb/c mice fed with modified diet either with EPA or with DHA for 3 weeks, spleen lymphocytes decreased their production of TNF-[alpha] only in the diet with EPA, but not in the diet with DHA. When approaching the effect of microalgae fatty acids, a study carried out in cell lines of macrophages exposed to LPS and added with extracts of different microalgae showed a significant decrease of the TNF-[alpha] compared against the control cultures. A study by Sierra et al. reported that only EPA enriched diet was able to decrease IL-12 concentrations [45].

In this study, the percentage of TCD4+ cells producing IL-17A showed a significant increase in both strains. IL-17A is known as an inflammatory cytokine whose main function is exerted on myeloid and mesenchymal cells by inducing the expression of granulocyte colony-stimulating factor (G-CSF), IL-6, and other chemokines, which increase granulopoiesis and recruit neutrophils into the site of infection [46]. Vigerust et al. also reported that, after supplementation with fish oil and krill oil enriched diets, IL-17A was shown to be increased only in the fish oil group [40], and these results agree with our findings.

The supplementation with n-3 fatty acids in lyophilized form and that are added in the food showed a significantly higher percentage of TCD4+ cells producing IL-10 in both strains. IL-4 is produced by type 2 T cells (Th2), basophils, and mast cells. It has anti-inflammatory function by blocking the synthesis of IL-1, TNF-[alpha], and IL-6. In addition, it promotes the proliferation and differentiation of B lymphocytes and is considered a potent inhibitor of apoptosis [47]. IL-10 is also known as cytokine synthesis inhibiting factor (CSIF) and can inhibit the synthesis of proinflammatory cytokines by T lymphocytes and macrophages. It also regulates the growth and differentiation of B lymphocytes, NK cytotoxic and helper T cells, mast cells, granulocytes, dendritic cells, keratinocytes, and endothelial cells [48]. A study in Wistar rats fed with a fish oil enriched diet was found to show lower concentrations of IL-4 and IL-5 at the alveolar level compared to control [41]. Also, Sierra et al. [45] showed similar results in Balb/c mice. Our findings agree with Li et al. [42] who demonstrated that fish oil supplementation decreased IL-10; however, Sierra et al. reported that only EPA supplementation was able to increase IL-10 concentrations [45]. These results suggest that the consumption of microalgae fatty acids has attenuating effects of systemic inflammation even in chronic diseases and not only in acute states of inflammation.

TGF-[beta] is a cytokine with pleiotropic functions in hematopoiesis, angiogenesis, cell proliferation, differentiation, migration, and apoptosis. It has a strong antiinflammatory action but may increase some immune functions. Thus, in knock-out mice for TGF-[micro] they show defects in regulatory T lymphocytes, which generates an extensive inflammation with abundant proliferation of T lymphocytes and differentiation of CD4+ in Th1 and Th2 lymphocytes [49]. In our study both strains showed higher percentage of TCD4+ cells producing TGF-[beta] in the LY group when compared with BL; however, they were much lower than those for the SAT group so we could agree that microalgae fatty acids have a positive effect on TGF-[beta] expression. A study with apoE-deficient mice infused with angiotensin and treated with EPA and DHA orally for 3 weeks showed that the TGF-[beta] gene expression was significantly decreased in the EPA group and DHA compared to untreated mice [50]. In contrast, a study on Wistar rats [51] exposed to LPS during gestation and whose offspring were supplemented with fish oil showed that TGF-[beta]] concentrations were increased as compared to controls [52].

IL-6 release is induced by IL-1 and TNF-a. It is involved in the production of immunoglobulins and in the differentiation of active B lymphocytes and plasma cells, modulates hematopoiesis, and is responsible, together with IL-1, for the synthesis of acute phase liver proteins like fibrinogen [53]. Percentage of TCD4+ cells producing IL-6 was shown to be significantly increased in LY group from both strains; however SAT group reported the highest IL-6 concentrations; we suggest that microalgae fatty acids could have a protective effect against IL-6 expression. Two studies report decreasing concentrations of IL-6 when using fish oil: one was conducted in mice [42] and the other in overweight pregnant women [43]. Additionally, a study by Robertson et al. [26] in macrophage cell line cultures treated with microalgae extracts showed a significant decrease in IL-6.

Regarding the effects of the consumption of coconut oil as a source of saturated fat, there is controversy about the properties of coconut oil, and this is because, despite being a source of saturated fat, several studies have shown that it has anti-inflammatory properties [54, 55]; in this study, the consumption of coconut oil showed a high percentage of TCD4+ cells producing Th1 and Th2 type cytokines which is consistent with other studies [56].

5. Conclusion

The results provided in this study show evidence that supplementation with n-3 fatty acids obtained from microalgae improves the inflammatory profile in general by reducing the secretion of many cytokines. Therefore, these results suggest that microalgae extracts maybe considered as an antiinflammatory strategy against different chronic diseases.

https://doi.org/10.1155/2018/4765358

Conflicts of Interest

The authors declare that there are no conflicts of interest regarding the publication of this paper.

Acknowledgments

Microalgae n-3 fatty acid lyophilized and modified diet was provided by Biotecnologia Mexicana de Microalgas SA de CV. GPLE is a CONACyT fellowship program member.

References

[1] Diagnosis and Classification of Diabetes Mellitus, vol. 37, supplement 1, Diabetes Care, 2014.

[2] T. Shamah-Levy, L. Cuevas-Nasu, J. Rivera-Dommarco, and M. Hernandez-Avila, Encuesta Nacional de Nutricion y Salud de Medio Camino 2016 (ENSANUT MC 2016), Inf Final Result Recuper, 2016.

[3] M. Hernandez-Avila, J. P. Gutierrez, and N. Reynoso-Noveron, "Diabetes mellitus en Mexico. El estado de la epidemia," Salud Publica de Mexico, vol. 55, pp. s129-s136, 2013.

[4] K. Esposito, R. Marfella, and D. Giugliano, "Stress hyperglycemia, inflammation, and cardiovascular events [18]," Diabetes Care, vol. 26, no. 5, pp. 1650-1651, 2003.

[5] T. A. Mori, R. J. Woodman, V. Burke, I. B. Puddey, K. D. Croft, and L. J. Beilin, "Effect of eicosapentaenoic acid and docosahexaenoic acid on oxidative stress and inflammatory markers in treated-hypertensive type 2 diabetic subjects," Free Radical Biology & Medicine, vol. 35, no. 7, pp. 772-781, 2003.

[6] V. M. Montori, A. Farmer, P. C. Wollan, and S. F. Dinneen, "Fish oil supplementation in type 2 diabetes: a quantitative systematic review," Diabetes Care, vol. 23, no. 9, pp. 1407-1415, 2000.

[7] S. L. Connor and W. E. Connor, "Are fish oils beneficial in the prevention and treatment of coronary artery disease?" American Journal of Clinical Nutrition, vol. 66, no. 4, pp. 1020S-1031S, 1997.

[8] T. A. Mori and L. J. Beilin, "Long-chain omega 3 fatty acids, blood lipids and cardiovascular risk reduction," Current Opinion in Lipidology, vol. 12, no. 1, pp. 11-17, 2001.

[9] P. C. Calder, "Immunomodulation by omega-3 fatty acids," Prostaglandins, Leukotrienes and Essential Fatty Acids, vol. 77, no. 5-6, pp. 327-335, 2007.

[10] B. Liang, S. Wang, Y.-J. Ye et al., "Impact of postoperative omega-3 fatty acid-supplemented parenteral nutrition on clinical outcomes and immunomodulationsi in colorectal cancer patients," World Journal of Gastroenterology, vol. 14, no. 15, pp. 2434-2439, 2008.

[11] J. E. Teitelbaum and W. Allan Walker, "Review: The role of omega 3 fatty acids in intestinal inflammation," The Journal of Nutritional Biochemistry, vol. 12, no. 1, pp. 21-32, 2001.

[12] J. P. Schuchardt, I. Schneider, and H. Meyer, "Incorporation of EPA and DHA into plasma phospholipids in response to different omega-3 fatty acid formulations--a comparative bioavailability study of fish oil vs. krill oil," Lipids in Health and Disease, vol. 10, article 145, 2011.

[13] D. Y. Oh, S. Talukdar, E. J. Bae et al., "GPR120 is an omega-3 fatty acid receptor mediating potent anti-inflammatory and insulin-sensitizing effects," Cell, vol. 142, no. 5, pp. 687-698, 2010.

[14] M. Arita, F. Bianchini, J. Aliberti et al., "Stereochemical assignment, antiinflammatory properties, and receptor for the omega3 lipid mediator resolvin E1," The Journal of Experimental Medicine, vol. 201, no. 5, pp. 713-722, 2005.

[15] R. Wall, R. P. Ross, G. F. Fitzgerald, and C. Stanton, "Fatty acids from fish: the anti-inflammatory potential of long-chain omega-3 fatty acids," Nutrition Reviews, vol. 68, no. 5, pp. 280-289, 2010.

[16] J. A. Hall, R. C. Wander, J. L. Gradin, S. H. Du, and D. E. Jewell, "Effect of dietary n-6-to-n-3 fatty acid ratio on complete blood and total white blood cell counts, and T-cell subpopulations in aged dogs," Am J Vet Res, vol. 60, pp. 319-327,1999.

[17] C. A. Hudert, K. H. Weylandt, Y. Lu et al., "Transgenic mice rich in endogenous omega-3 fatty acids are protected from colitis," Proceedings of the National Acadamy of Sciences of the United States of America, vol. 103, no. 30, pp. 11276-11281, 2006.

[18] G. P. Lim, F. Calon, T. Morihara et al., "A diet enriched with the omega-3 fatty acid docosahexaenoic acid reduces amyloid burden in an aged Alzheimer mouse model," The Journal of Neuroscience, vol. 25, no. 12, pp. 3032-3040, 2005.

[19] N. S. Kalupahana, K. Claycombe, S. J. Newman et al., "Eicosapentaenoic acid prevents and reverses insulin resistance in high-fat diet-induced obese mice via modulation of adipose tissue inflammation," Journal of Nutrition, vol. 140, no. 11, pp. 1915-1922, 2010.

[20] J. M. Bourre, "Roles of unsaturated fatty acids (especially omega-3 fatty acids) in the brain at various ages and during ageing," J Nutr Health Aging [Internet], vol. 8, no. 3, pp. 163-174, 2004.

[21] A. Valenzuela B and R. Valenzuela B, "Acidos grasos omega-3 en la nutricion jcomo aportarlos?" Revista chilena de nutricion, vol. 41, no. 2, pp. 205-211, 2014.

[22] I. Khozin-Goldberg, S. Leu, and S. Boussiba, "Microalgae as a source for VLC-PUFA production," Subcellular Biochemistry, vol. 86, pp. 471-510, 2016.

[23] C. J. Shepherd and A. J. Jackson, "Global fishmeal and fish-oil supply: Inputs, outputs and marketsa," Journal of Fish Biology, vol. 83, no. 4, pp. 1046-1066, 2013.

[24] R. Robertson, F. Guiheneuf, DB. Stengel, G. Fitzgerald, P. Ross, and C. Stanton, "Algae-derived Polyunsaturated fatty acids: implications for human health," in Polyunsaturated Fatty Acifs: Sources, Antioxidant Properties and Health Benefits, 2017

[25] J. G. Bell and J. R. Sargent, "Arachidonic acid in aquaculture feeds: Current status and future opportunities," Aquaculture, vol. 218, no. 1-4, pp. 491-499, 2003.

[26] R. C. Robertson, F. Guiheneuf, B. Bahar et al., "The anti-inflammatory effect of algae-derived lipid extracts on lipopolysaccharide (LPS)-stimulated human THP-1 macrophages," Marine Drugs, vol. 13, no. 8, pp. 5402-5424, 2015.

[27] R. Deng and T.-J. Chow, "Hypolipidemic, antioxidant, and antiinflammatory activities of microalgae spirulina," Cardiovascular Therapeutics, vol. 28, no. 4, pp. e33-e45, 2010.

[28] E. Talero, S. Garcia-Maurino, J. Avila-Roman, A. RodriguezLuna, A. Alcaide, and V. Motilva, "Bioactive compounds isolated from microalgae in chronic inflammation and cancer," Marine Drugs, vol. 13, no. 10, pp. 6152-6209, 2015.

[29] I. M. Arciniega-Martinez, R. Campos-Rodriguez, M. E. DragoSerrano, L. E. Saenchez-Torres, T. R. Cruz-Hernaendez, and A. A. Resendiz-Albor, "Modulatory Effects of Oral Bovine Lactoferrin on the IgA Response at Inductor and Effector Sites of Distal Small Intestine from BALB/c Mice," Archivum Immunologiae et Therapia Experimentalis, vol. 64, no. 1, pp. 5763, 2016.

[30] P. Zhuang, W. Wang, Y. Zhang, and J. Jiao, "Long-term dietary EPA or DHA supplementation do not ameliorate obesity but improve glucose homeostasis via gut-adipose axis in already obese mice," FASEB J, vol. 31, no. 1, pp. 971-11, 2017.

[31] P. Flachs, V. Mohamed-Ali, O. Horakova et al., "Polyunsaturated fatty acids of marine origin induce adiponectin in mice fed a high-fat diet," Diabetologia, vol. 49, no. 2, pp. 394-397, 2006.

[32] J. M. Monk, D. M. Liddle, and A. A. De Boer, "Fish-oil-derived n-3 PUFAs reduce inflammatory and chemotactic adipokinemediated cross-talk between co-cultured murine splenic CD8+ T cells and adipocytes," Journal of Nutrition, vol. 145, no. 4, pp. 829-838, 2015.

[33] F. de Jesus Diaz-Resendiz, K. Franco-Paredes, A. G. Martinez-Moreno, A. Lopez-Espinoza, and V G. Aguilera-Cervantes, Efectos de variables ambientales sobre la ingesta de alimento en ratas: Una revision historico-conceptual, vol. 8, Univ Psychol, 2009.

[34] L. Marano, R. Porfidia, M. Pezzella et al., "Clinical and immunological impact of early postoperative enteral immunonutrition after total gastrectomy in gastric cancer patients: a prospective randomized study," Annals of Surgical Oncology, vol. 20, no. 12, pp. 3912-3918, 2013.

[35] A. M. Rizzo, P. A. Corsetto, G. Montorfano et al., "Comparison between the AA/EPA ratio in depressed and non depressed elderly females: Omega-3 fatty acid supplementation correlates with improved symptoms but does not change immunological parameters," Nutrition Journal, vol. 11, no. 1, article no. 82, 2012.

[36] V. R. Mukaro, M. Costabile, K. J. Murphy, C. S. Hii, P. R. Howe, and A. Ferrante, "Leukocyte numbers and function in subjects eating n-3 enriched foods: Selective depression of natural killer cell levels," Arthritis Research & Therapy, vol. 10, no. 3, article no. R57, 2008.

[37] G. C. Baccan, O. Hernandez, L. E. Diaz et al., "Changes in lymphocyte subsets and functions in spleen from mice with high fat diet-induced obesity," Proceedings of the Nutrition Society, vol. 72, no. OCE1, 2013.

[38] K. Schroder, P. J. Hertzog, T. Ravasi, and D. A. Hume, "Interferon-gamma: an overview of signals, mechanisms and functions," J Leukoc Biol, vol. 75, no. 2, pp. 163-189, 2004.

[39] C. L. Langrish, B. S. McKenzie, N. J. Wilson, R. De Waal Malefyt, R. A. Kastelein, and D. J. Cua, "IL-12 and IL-23: master regulators of innate and adaptive immunity," Immunological Reviews, vol. 202, pp. 96-105, 2004.

[40] N. F. Vigerust, B. Bjorndal, P. Bohov, T. Brattelid, A. Svardal, and R. K. Berge, "Krill oil versus fish oil in modulation of inflammation and lipid metabolism in mice transgenic for TNF-[alpha]," European Journal of Nutrition, vol. 52, no. 4, pp. 1315-1325, 2013.

[41] R. A. N. Xavier, K. V de Barros, I. S. de Andrade, Z. Palomino, D. E. Casarini, and V. L. F. Silveira, "Protective effect of soybean oil or fish oil-rich diets on allergic airway inflammation," Journal of Inflammation Research, vol. 9, pp. 79-89, 2016.

[42] C. C. Li, H. T. Yang, Y. C. Hou, Y. S. Chiu, and W. C. Chiu, "Dietary fish oil reduces systemic inflammation and ameliorates sepsis-induced liver injury by up-regulating the peroxisome proliferator-activated receptor gamma-mediated pathway in septic mice," The Journal of Nutritional Biochemistry, vol. 25, pp. 19-25, 2014.

[43] M. Ebrahimi-Mameghani, Z. Sadeghi, M. Abbasalizad Farhangi, E. Vaghef-Mehrabany, and S. Aliashrafi, "Glucose homeostasis, insulin resistance and inflammatory biomarkers in patients with non-alcoholic fatty liver disease: Beneficial effects of supplementation with microalgae Chlorella vulgaris: A double-blind placebo-controlled randomized clinical trial," Clinical Nutrition, vol. 36, no. 4, pp. 1001-1006, 2017

[44] M. Haghiac, X.-H. Yang, L. Presley et al., "Dietary omega-3 fatty acid supplementation reduces inflammation in obese pregnant women: A randomized double-blind controlled clinical trial," PLoS ONE, vol. 10, no. 9, Article ID e0137309, 2015.

[45] S. Sierra, F. Lara-Villoslada, M. Comalada, M. Olivares, and J. Xaus, "Dietary eicosapentaenoic acid and docosahexaenoic acid equally incorporate as decosahexaenoic acid but differ in inflammatory effects," Nutrition Journal, vol. 24, no. 3, pp. 245-254, 2008.

[46] Y. F. Talmas-Rohana, "Interleucina 17, funciones biologicas y su receptor," Reb, vol. 31, no. 1, pp. 3-9, 2012.

[47] A. E. Kelly-Welch, E. M. Hanson, M. R. Boothby, and A. D. Keegan, "Interleukin-4 and interleukin-13 signaling connections maps," Science, vol. 300, no. 5625, pp. 1527-1528, 2003.

[48] K. W. Moore, R. de Waal Malefyt, R. L. Coffman, and A. O'Garra, "Interleukin-10 and the interleukin-10 receptor," Annual Review of Immunology, vol. 19, pp. 683-765, 2001.

[49] E. Gonzalo-Gil and M. Galindo-Izquierdo, "Papel del factor de crecimiento transformador-beta (TGF-[micro]) en la fisiopatologia de la artritis reumatoide," Reumatologia CUnica, vol. 10, no. 3, pp. 174-179, 2014.

[50] T. Yoshihara, K. Shimada, K. Fukao et al., "Omega 3 polyunsaturated fatty acids suppress the development of aortic aneurysms through the inhibition of macrophage-mediated inflammation," Circulation Journal, vol. 79, no. 7, pp. 1470-1478, 2015.

[51] A. Rabinovitch and W. L. Suarez-Pinzon, "Cytokines and their roles in pancreatic islet [micro]-cell destruction and insulindependent diabetes mellitus," Biochemical Pharmacology, vol. 55, no. 8, pp. 1139-1149, 1998.

[52] J. J. Fortunato, N. da Rosa, A. O. Martins Laurentino et al., "Effects of w-3 fatty acids on stereotypical behavior and social interactions in Wistar rats prenatally exposed to lipopolysaccarides," Nutrition Journal, vol. 35, pp. 119-127, 2017

[53] A. D. Pradhan, J. E. Manson, N. Rifai, J. E. Buring, and P. M. Ridker, "C-reactive protein, interleukin 6, and risk of developing type 2 diabetes mellitus," The Journal of the American Medical Association, vol. 286, no. 3, pp. 327-334, 2001.

[54] H. Van Der Tempel, J. E. Tulleken, P. C. Limburg, F. A. J. Muskiet, and M. H. Van Rijswijk, "Effects of fish oil supplementation in rheumatoid arthritis," Annals of the Rheumatic Diseases, vol. 49, no. 2, pp. 76-80,1990.

[55] R. J. Goldberg and J. Katz, "A meta-analysis of the analgesic effects of omega-3 polyunsaturated fatty acid supplementation for inflammatory joint pain," PAIN,vol. 129, no. 1-2, pp. 210-223, 2007.

[56] S. Intahphuak, P. Khonsung, and A. Panthong, "Antiinflammatory, analgesic, and antipyretic activities of virgin coconut oil," Pharmaceutical Biology, vol. 48, no. 2, pp. 151-157, 2010.

L. E. Gutierrez-Pliego (iD), (1) B. E. Martinez-Carrillo (iD), (1) A. A. Resendiz-Albor (iD), (2) I. M. Arciniega-Martinez (iD), (2) J. A. Escoto-Herrera (iD), (1) C. A. Rosales-Gomez (iD), (1) and R. Valdes-Ramos (iD) (1)

(1) Laboratorio de Investigacion en Nutricion, Facultad de Medicina, Universidad Autonoma del Estado de Mexico, Paseo Tollocany Venustiano Carranza s/n, Col. Universidad, 50180 Toluca, MEX, Mexico

(2) Laboratorio de Inmunidad de Mucosas, Seccion de Investigacion y Posgrado, Escuela Superior de Medicina, Instituto Politecnico Nacional, Av. Plan de San Luis S/N, Colonia Casco de Santo Tomas, Miguel Hidalgo, 11350 Ciudad de Mexico, Mexico

Correspondence should be addressed to R. Valdes-Ramos; rvaldesr@uaemex.mx

Received 29 November 2017; Revised 1 February 2018; Accepted 21 February 2018; Published 1 April 2018

Academic Editor: Gerhard M. Kostner

Caption: Figure 1: Experimental groups for both strains.

Caption: Figure 2: Summary of major findings about microalgae fatty acids supplementation in diabetic and healthy mice.
Table 1: Nutrient composition of study groups' diet.

RC group                        LY group

Protein, %             23.9     Protein, %             23.9
Starch, %              31.9     Starch, %              31.9
Glucose, %             0.22     Glucose, %             0.22
Fiber (crude), %       5.10     Fiber (crude), %       5.10
Cholesterol, ppm       200      Cholesterol, ppm       200
EPA + DHA, %           0.2      EPA + DHA, %           0.2

Metabolizable energy:           Metabolizable energy:
3.02 kcal/g                     3.02 kcal/g + 0.09  kcal/mg of
                                lyophilizedfatty acids *

SAT group                       MD group

Protein, %             23.9     Protein, %             23.9
Starch, %              31.9     Starch, %              31.9
Glucose, %             0.22     Glucose, %             0.22
Fiber (crude), %       5.10     Fiber (crude), %       5.10
Cholesterol, ppm       200      Cholesterol, ppm       200
EPA + DHA, %           0.2      EPA + DHA', %          2.0

Metabolizable energy:           Metabolizable energy:
3.02 kcal/g + 0.09              3.02 kcal/g
kcal/mg of
coconut oil

* Microalgae source.

Table 2: Effect of supplementation with EPA and DHA extracted
from microalgae on body mass index, blood glucose, food, and water
consumption in db/db and CD1 mice.

                                         8 weeks old
                                  BL                      RC
                           Mean [+ or -] SD        Mean [+ or -] SD
                                 n = 6                   n = 6

db/db

BMI g/[cm.sup.2]           573 [+ or -] 4.9        61.2 [+ or -] 2.5
Glucose mg/dL            293.8 [+ or -] 131.0    551.8 [+ or -] 83.7 *
Food intake, g/week        34.3 [+ or -] 2.1       32.9 [+ or -] 0.3
Water intake, mL/week     64.8 [+ or -] 11.8       78.2 [+ or -] 7.2

CD1

BMI g/[cm.sup.2]           29.6 [+ or -] 2.0       31.4 [+ or -] 1.9
Glucose mg/dL             126.2 [+ or -] 18.6      119.3 [+ or -] 13
Food intake, g/week        54.6 [+ or -] 1.7       58.3 [+ or -] 5.6
Water intake, mL/week      63 [+ or -] 13.1        673 [+ or -] 3.2

                                        16 weeks old
                                  MD                     LY
                           Mean [+ or -] SD       Mean [+ or -] SD
                                n = 6                  n = 6

db/db

BMI g/[cm.sup.2]          65 [+ or -] 1.7 *      62.2 [+ or -] 1.5
Glucose mg/dL             505 [+ or -] 74 *     525.5 [+ or -] 51 *
Food intake, g/week      27.6 [+ or -] 1.0 *    37.5 [+ or -] 0.8 *
Water intake, mL/week    81.4 [+ or -] 10.4 *    79.6 [+ or -] 1.8

CD1

BMI g/[cm.sup.2]          32.2 [+ or -] 3.1      34.4 [+ or -] 4.1
Glucose mg/dL            127.5 [+ or -] 18.7    109.7 [+ or -] 13.3
Food intake, g/week         42.7[+ or -]5         61 [+ or -] 1.9
Water intake, mL/week     53.9 [+ or -] 1.3     89.8 [+ or -] 6.1 *

                                 SAT
                           Mean [+ or -] SD
                                n = 6

db/db

BMI g/[cm.sup.2]          59.7 [+ or -] 2.3
Glucose mg/dL             580.7 [+ or -] 22*
Food intake, g/week       36.3 [+ or -] 2.4
Water intake, mL/week     67.2 [+ or -] 8.3

CD1

BMI g/[cm.sup.2]          34.2 [+ or -] 2.4
Glucose mg/dL            106.8 [+ or -] 12.4
Food intake, g/week      73.3 [+ or -] 20.8 *
Water intake, mL/week    93.2 [+ or -] 4.8 *

Data are presented as means [+ or -] standard deviations. One-way
ANOVA* for comparison of differences between BL group at 8 weeks
versus all the groups at 16 weeks. p value was significant at <0.05.

Table 3: Effect of supplementation with EPA and DHA extracted
from microalgae in lymphocytes populations in db/db and CD1 mice.

              8 weeks old
                   BL                     RC
            Mean [+ or -] SD       Mean [+ or -] SD
                 n = 6                  n = 6

db/db

CD3+, %    71.8 [+ or -] 0.5     70.4 [+ or -] 0.8 *
CD4+, %    62.0 [+ or -] 0.2      62.1 [+ or -] 0.5
CD8+, %    30.0 [+ or -] 1.0      29.3 [+ or -] 1.0

cd1

CD3+, %    74.7 [+ or -] 0.02    77.8 [+ or -] 0.04 *
CD4+, %    67.4 [+ or -] 0.2      67.5 [+ or -] 0.05
CD8+, %    21.4 [+ or -] 0.05    24.8 [+ or -] 0.01 *

                        16 weeks old
                   MD                     LY
            Mean [+ or -] SD       Mean [+ or -] SD
                 n = 6                  n = 6

db/db

CD3+, %   69.8 [+ or -] 0.4 *    70.7 [+ or -] 0.4 *
CD4+, %   60.6 [+ or -] 0.3 *     61.8 [+ or -] 0.9
CD8+, %   24.9 [+ or -] 0.3 *    28.4 [+ or -] 0.2*

cd1

CD3+, %   70.6 [+ or -] 0.03 *   72.4 [+ or -] 0.01 *
CD4+, %    70 [+ or -] 0.04 *     67.6 [+ or -] 0.03
CD8+, %   17.4 [+ or -] 0.1 *    24.2 [+ or -] 0.03 *

                  SAT
            Mean [+ or -] SD
                 n = 6

db/db

CD3+, %   68.3 [+ or -] 0.2 *
CD4+, %    65 [+ or -] 0.5 *
CD8+, %    23 [+ or -] 0.1 *

cd1

CD3+, %   69.5 [+ or -] 0.06 *
CD4+, %   72.3 [+ or -] 0.4 *
CD8+, %   19.9 [+ or -] 0.02 *

Data are pas means [+ or -] standard deviations.
One-way ANOVA* for comparison of differences between BL group
at 8 weeks versus all the groups at 16 weeks. p value was significant
at <0.05.

Table 4: Effect of supplementation with EPA and DHA fatty acids
extracted from microalgae on Th1 and Th2 cytokines in db/db.

                      8 weeks old
db/db mice                 BL                     RC
                    Mean [+ or -] SD       Mean [+ or -] SD
                         n = 6                  n = 6

Th1 % TCD4+/

IFN-[gamma]       22.5 [+ or -] 0.4      11.5 [+ or -] 0.4 *
TNF-[alpha]       10.2 [+ or -] 0.4      7.7 [+ or -] 0.4 *
IL-12             1.2 [+ or -] 0.1       12.4 [+ or -] 0.5 *
IL-17A            1.4 [+ or -] 0.1       4.2 [+ or -] 0.3 *

Th2 % TCD4+/

IL-4              1.8 [+ or -] 0.08      8.2 [+ or -] 0.3 *
IL-5              19.9 [+ or -] 0.8      9.4 [+ or -] 0.4 *
IL-6              1.5 [+ or -] 0.1       3.1 [+ or -] 0.4 *
IL-10             1.44 [+ or -] 0.2      16.0 [+ or -] 1.3 *
TGF-[beta]        3.2 [+ or -] 0.4       2.1 [+ or -] 0.05 *

                                16 weeks old
db/db mice               MD LY
                    Mean [+ or -] SD       Mean [+ or -] SD
                         n = 6                  n = 6

Th1 % TCD4+/

IFN-[gamma]       7.2 [+ or -] 0.5 *     2.1 [+ or -] 0.4 *
TNF-[alpha]       8.8 [+ or -] 0.4 *     2.4 [+ or -] 0.4 *
IL-12             10.2 [+ or -] 0.5 *    6.8 [+ or -] 0.1 *
IL-17A            3.7 [+ or -] 0.1 *     11.4 [+ or -] 0.3 *

Th2 % TCD4+/

IL-4              6.6 [+ or -] 0.3 *     1.1 [+ or -] 0.1 *
IL-5              7.5 [+ or -] 0.6 *     2.1 [+ or -] 0.2 *
IL-6              4.5 [+ or -] 0.2 *     4.0 [+ or -] 0.1 *
IL-10             7.3 [+ or -] 0.2 *     2.7 [+ or -] 0.08 *
TGF-[beta]        5.5 [+ or -] 0.1 *     4.9 [+ or -] 0.09 *

db/db mice                SAT
                    Mean [+ or -] SD
                         n = 6

Th1 % TCD4+/

IFN-[gamma]       15.7 [+ or -] 0.4 *
TNF-[alpha]       1.4 [+ or -] 0.4 *
IL-12             3.6 [+ or -] 0.5 *
IL-17A            11.4 [+ or -] 0.3 *

Th2 % TCD4+/

IL-4              10.3 [+ or -] 0.2 *
IL-5              6.5 [+ or -] 0.2 *
IL-6              14.8 [+ or -] 0.2 *
IL-10             14.5 [+ or -] 0.2 *
TGF-[beta]        7.1 [+ or -] 0.1 *

Data are presented as means [+ or -] standard deviations of
percentage of TCD4+ cells producing cytokines. One-way ANOVA*
for comparison of differences between BL group at 8 weeks versus
all the groups at 16 weeks. p value was significant at <0.05.

Table 5: Effect of supplementation with EPA and DHA fatty acids
extracted from microalgae on Th1 and Th2 cytokines in CD1.

                  8 weeks old
CD1 mice                BL                     RC
                 Mean [+ or -] SD       Mean [+ or -] SD
                      n = 6                  n = 6

Th1

% TCD4+/
IFN-[gamma]    1.8 [+ or -] 0.18      1.2 [+ or -] 0.09 *
TNF-[alpha]    2.5 [+ or -] 0.13      1.5 [+ or -] 0.1 *
IL-12          2.8 [+ or -] 0.1       1.7 [+ or -] 0.08 *
IL-17A         1.6 [+ or -] 0.08      1.8 [+ or -] 0.08 *

Th2

% TCD4+/
IL-4           2.6 [+ or -] 0.1       1.8 [+ or -] 0.09 *
IL-5           2.1 [+ or -] 0.1       1.4 [+ or -] 0.1 *
IL-6           1.8 [+ or -] 0.1       1.7 [+ or -] 0.08
IL-10          1.6 [+ or -] 0.08      1.5 [+ or -] 0.1
TGF-[beta]     1.5 [+ or -] 0.06      1.3 [+ or -] 0.8 *

                             16 weeks old
CD1 mice                MD                     LY
                 Mean [+ or -] SD       Mean [+ or -] SD
                      n = 6                  n = 6

Th1

% TCD4+/
IFN-[gamma]    2.7 [+ or -] 0.1 *     1.0 [+ or -] 0.1 *
TNF-[alpha]    2.6 [+ or -] 0.1       1.4 [+ or -] 0.09 *
IL-12          2.4 [+ or -] 0.09 *    1.3 [+ or -] 0.08 *
IL-17A         1.8 [+ or -] 0.1 *     4.1 [+ or -] 0.08 *

Th2

% TCD4+/
IL-4           2.4 [+ or -] 0.1       1.6 [+ or -] 0.1 *
IL-5           2.6 [+ or -] 0.1 *     1.9 [+ or -] 0.1
IL-6           1.7 [+ or -] 0.1       1.5 [+ or -] 0.09 *
IL-10          2.2 [+ or -] 0.1       1.2 [+ or -] 0.1
TGF-[beta]     2.3 [+ or -] 0.07 *    1.5 [+ or -] 0.08

CD1 mice               SAT
                 Mean [+ or -] SD
                      n = 6

Th1

% TCD4+/
IFN-[gamma]    4.3 [+ or -] 0.1 *
TNF-[alpha]    4.6 [+ or -] 0.07 *
IL-12          4.4 [+ or -] 0.08 *
IL-17A         3.6 [+ or -] 0.09 *

Th2

% TCD4+/
IL-4           4.4 [+ or -] 0.1 *
IL-5           3.4 [+ or -] 0.1 *
IL-6           2.6 [+ or -] 0.07 *
IL-10          2.9 [+ or -] 1.6
TGF-[beta]     5.1 [+ or -] 0.1 *

Data are presented as means [+ or -] standard deviations
of percentage of TCD4+ cells producing cytokines. One-way
ANOVA* for comparison of differences between BL group at
8 weeks versus all the groups at 16 weeks. p value was
significant at <0.05.
COPYRIGHT 2018 Hindawi Limited
No portion of this article can be reproduced without the express written permission from the copyright holder.
Copyright 2018 Gale, Cengage Learning. All rights reserved.

Article Details
Printer friendly Cite/link Email Feedback
Title Annotation:Research Article
Author:Gutierrez-Pliego, L.E.; Martinez-Carrillo, B.E.; Resendiz-Albor, A.A.; Arciniega-Martinez, I.M.; Esc
Publication:Journal of Lipids
Date:Jan 1, 2018
Words:7618
Previous Article:Intracellular and Plasma Membrane Events in Cholesterol Transport and Homeostasis.
Next Article:Comparison of Oil Content and Fatty Acids Profile of Western Schley, Wichita, and Native Pecan Nuts Cultured in Chihuahua, Mexico.
Topics:

Terms of use | Privacy policy | Copyright © 2021 Farlex, Inc. | Feedback | For webmasters |