Fatty acids of follicular fluid phospholipids and triglycerides display distinct association with IVF outcomes.

RESEARCH QUESTION: Are triglyceride fatty acids in the follicular fluid associated with either follicular fluid phospholipid fatty acids or IVF outcomes and, if so, how are they associated? DESIGN: In a prospective cross-sectional study, 70 women undergoing intracytoplasmic sperm injection were recruited. Follicular fluid phospholipids and triglycerides were separated by thin-layer chromatography. Fatty acids were measured using gas-liquid chromatography and flame ionization detection system. RESULTS: Significant differences in fatty acid composition were observed between follicular fluid phospholipid and triglyceride fractions. Phospholipid stearic acid and n-3 polyunsaturated fatty acids, particularly alpha-linolenic acid, were negatively associated with the number of mature oocytes and cleaved embryos, whereas arachidonic acid was in direct correlation with cleavage rate per IVF cycle (ß = 0.325, P = 0.022). In the case of triglyceride fraction, total monounsaturated fatty acids, oleic acid in particular, displayed significantly positive associations with the number of oocytes (ß = 0.261, P = 0.043) and embryos (ß = 0.310, P = 0.018). Furthermore, cleavage rate correlated inversely with palmitic acid (ß = -0.359, P = 0.007) and directly with pentadecanoic acid (ß = 0.378, P = 0.005). Most of these associations, however, were not independent of predictive fatty acids belonging to phospholipid fraction, according to multivariate analysis. CONCLUSIONS: Fatty acid compositions of phospholipid and triglyceride fractions from human follicular fluid differentially correlate with IVF cycle parameters.

Human charcoal-stripped serum supplementation enhances both the stearoyl-coenzyme a desaturase 1 activity of cumulus cells and the in vitro maturation of oocytes.

Obtaining a better outcome in assisted reproductive technology remains to be attained. In the case of in vitro fertilization (IVF), oocyte maturity is paramount for achieving a successful pregnancy. Maternal serum supplementation of in vitro maturation (IVM) medium can increase the rate of oocyte IVM. The aim of the present study was to compare the effect of whole and charcoal-stripped serum supplementation on IVM and the activity index of stearoyl-coenzyme A desaturase 1 (SCD1) in cumulus cells enclosing the oocyte as a molecular indicator of oocyte quality. Cumulus cells and germinal vesicle immature oocytes were collected from 76 women with polycystic ovarian syndrome during an IVF cycle. Serum samples were pooled from healthy women and were applied as whole or charcoal-stripped serum supplements. SCD1 expression and activity were measured by quantitative polymerase chain reaction (PCR) and gas-liquid chromatography, respectively. Charcoal-stripped serum at an amount of 10% showed a higher potency in increasing the SCD1 expression and activity index than whole serum (>1.5 fold, p < 0.001). An increase in the IVM rate was also observed in oocytes cultured in the presence of 10% charcoal-stripped serum compared to the control group (1.9 fold, p = 0.031). Therefore, charcoal-based lipid depletion as a simple and preparative strategy may increase the beneficial effect of serum supplementation in oocyte IVM culture.

The role of cholesterol-enriched diet and paraoxonase 1 inhibition in atherosclerosis progression.

Introduction: Atherosclerosis could be deemed as a chronic, progressive, and inflammatory disease. It has been well-documented that high-density lipoprotein (HDL) can reduce the risk of the atherosclerosis occurrence through exerting some anti-atherogenic mechanisms. In recent years, the strong evidence has suggested that paraoxonase 1 (PON1) may contribute to antioxidant properties of HDL. In the present study, the impact of a diet enriched with cholesterol and also the PON1 inhibition on atheroma formation and lipid profile has been investigated. Methods: In this study, 24 New Zealand rabbits were randomly assigned to three groups receiving standard diet, atherogenic diet, and atherogenic diet plus once daily intramuscular injection of nandrolone decanoate as the PON1 inhibitor. Triglyceride, cholesterol, HDL, and low-density lipoprotein (LDL) were determined and both cholesterol accumulation in aorta and fatty streak formation were evaluated. Results: The comparison of the results in three groups reveals that cholesterol level in the group received cholesterol-enriched diet plus once daily injection of PON1 inhibitor was higher than the groups received standard diet or atherogenic diet without PON1 inhibitor (P < 0.05). Furthermore, the percentage of atheroma with type-I lesions was equal to 75% compared with the group received atherogenic diet plus nandrolone at 30%. Additionally, the differences in fatty streak formation in aorta, as well as the right and left coronary arteries in three groups given show that the difference between groups receiving atherogenic diet and standard diet was significantly lower (P < 0.05) than the difference between groups receiving atherogenic diet plus PON1 inhibitor and standard diet. Conclusion: It can be concluded that lack of paraoxanase1 or even reduced the activity of this enzyme could accelerate the progression of fatty streak lesions toward advanced atherosclerotic lesions.

Fatty acids composition of aorta and saphenous vein tissues in patients with coronary artery diseases.

Introduction: Considering importance of fatty acids in developing coronary artery disease (CAD) and lack of information about saphenous vein which is commonly used as coronary arterial bypass, in this study we investigated differences in fatty acids composition between saphenous vein and aorta tissues in patients with CAD. Methods: Biopsy samples of aortic tissues and saphenous veins as well as blood samples were obtained form 42 patients with CAD. Fatty acids composition of the tissues was determined using gas chromatography and also serum lipid profile was evaluated by commercial kits. Results: Levels of palmitic acid (16:0) were significantly higher in aorta in compared with saphenous (P < 0.001). Also levels of most unsaturated fatty acids (16:1, 18:1n-9, 18:1t, 18:2t, 18:3 n-9 and 22:3n-3) were statistically higher in saphenous tissue than aorta tissue (P < 0.05). Mean levels of linoleic acid (18:2 n-6) was higher in aorta tissue in comparison with saphenous tissue (P = 0.01). We observed positive correlations between serum levels of LDL-C with elaidic acid and linoleic acid levels in saphenous. Evaluation of aorta tissue fatty acids revealed that palmitoleic acid (16:1) had positive and arachidonic and linoleic acids had negative correlations with serum HDL-C levels. Conclusion: Our results revealed difference between fatty acids composition of aorta and saphenous vein tissues and existence of correlations between the fatty acids levels with serum lipid profile. The saphenous vein had higher poly-unsaturated fatty acids in compared to aorta tissue and thus this vein is not at risk of atherosclerosis and can be used as coronary arterial bypass.

Secretory phospholipase-A2 and fatty acid composition in oral reactive lesions: a cross-sectional study.

BACKGROUND: Oral reactive lesions are the most common lesions of oral cavity. Phospholipases and fatty acids play key roles in the creation of inflammation by change in metabolic activities and production of lipid mediators. The aim of this study was to investigate the amount of secretory phospholipase-A2 (sPLA2) and difference of fatty acid pattern in oral reactive hyperplasia and adjacent normal appearing tissues in patients with oral reactive lesions. METHODS: Paired samples of oral hyperplastic (OH) and adjacent normal-appearing tissue of 45 patients were investigated in this study. The collected samples were analyzed with enzymatic spectrophotometric method in terms of the amount of sPLA2 and composition of fatty acids by gas-liquid chromatography method. RESULTS: The amount of sPLA2 (1.8-fold, p < 0.001), stearic acid (1.2-fold, p < 0.001), oleic acid (1.1-fold, p = 0.01), arachidonic acid (1.5-fold, p < 0.001) and docosahexaenoic acid (1.3-fold, p = 0.02) were increased, while the amount of palmitoleic acid (-45%, p < 0.001) and linoleic acid (-19%, p < 0.001) were reduced in the OH tissue samples. Furthermore, the results demonstrated significant associations between the type and location of tissue samples with monounsaturated fatty acids (MUFAs) and n-3 polyunsaturated fatty acids. Tissue samples from patients with inflammatory fibroepithelial hyperplasia showed relatively higher MUFAs and lower n-3 polyunsaturated fatty acids than other type of lesions. CONCLUSIONS: Localized changes in the sPLA2 activity and composition of fatty acid are associated with oral reactive hyperplasia and the type of pathological response. We suggest that sPLA2 activity and multiple type of fatty acids might be used as potential therapeutic target for oral reactive hyperplasia.

The effect of fish oil supplementation on serum phospholipid fatty acids profile during pregnancy: A double blind randomized controlled trial.

Omega-3 fatty acids (FAs) are essential unsaturated long-chain FAs necessary for proper health and growth. The objective of the authors in this study was to evaluate the effect of fish oil supplementation in pregnancy on maternal serum FA profiles. Participants (n = 150 pregnant women aged 18-35 years in Tabriz, Iran) were randomly assigned to receive either 1,000 mg fish oil supplements daily containing 120 mg docosahexanoic acid (DHA) and 180 mg eicosapentaenoic acid (EPA), or placebo from week 21 of pregnancy to delivery. The primary outcome measures were mean serum DHA and EPA proportion of total FAs at weeks 35-37 of pregnancy. Analyses were based on intention-to-treat. No significant differences were observed between the groups in consumption of fish and serum FAs levels at baseline. Fish oil supplementation significantly increased the mean DHA proportion of total FAs in the intervention compared to the placebo group at weeks 35-37 [adjusted Mean Difference (aMD) = 0.15; 95% CI 0.08-0.23]. The mean EPA proportion of total FAs also increased in the intervention group, but the difference between the groups was not significant (aMD = 0.04; 95% CI -0.01 to 0.08). The dietary recommendation for consumption of 1,000 mg/day fish oil supplements during pregnancy seems beneficial for better serum FA composition.

Fatty Acid Composition of Tissue Cultured Breast Carcinoma and the Effect of Stearoyl-CoA Desaturase 1 Inhibition.

PURPOSE: Stearoyl-CoA desaturase 1 (SCD1) is a novel therapeutic target in various malignancies, including breast cancer. The present study was designed to investigate the effect of the pharmacologic inhibition of SCD1 on fatty acid composition in tissue explant cultures of human breast cancer and to compare these effects with those in adjacent nonneoplastic breast tissue. METHODS: Paired samples of tumor and adjacent noncancerous tissue were isolated from 12 patients with infiltrating ductal breast cancer. Samples were explant cultured in vitro, exposed to the highly selective SCD1 inhibitor CAY10566, and examined for fatty acid composition by gas liquid chromatography. The cytotoxic and antigrowth effects were evaluated by quantification of lactate dehydrogenase release and by sulforhodamine B (SRB) measurement, respectively. RESULTS: Breast cancer tissue samples were found to have higher levels of monounsaturated fatty acids (MUFA) (p<0.001) and arachidonic acid (20:4n-6, p<0.001) and a lower level of linoleic acid (18:2n-6, p=0.02) than the normal-appearing breast tissues. While exhibiting no evident cytotoxicity, treatment with the SCD1 inhibitor, CAY10566 (0.1-1 µM), for 48 hours significantly increased 18:2n-6 levels in both the tumor and adjacent normal-appearing tissue (approximately 1.2 fold, p<0.05). However, the breast cancer tissue samples showed significant increases in the levels of MUFA and 20:4n-6 compared to the normal-appearing breast tissues (p<0.05). The SRB growth assay revealed a higher rate of inhibition with the SCD1 inhibitor in breast cancer tissues than in normal-appearing tissues (p<0.01, 41% vs. 29%). The SCD1 inhibitor also elevated saturated fatty acid (1.46-fold, p=0.001) levels only in the tumor tissue explant. CONCLUSION: The fatty acid composition and response to SCD1 inhibition differed between the explant cultures from breast cancer and the adjacent normal-appearing tissue. Altered fatty acid composition induced by SCD1 inhibition may also, in addition to Δ9 desaturation, modulate other reactions in de novo fatty acid synthesis and lipogenesis, and subsequently affect the overall survival and progression of breast cancer.

PPARγ agonist-induced alterations in Δ6-desaturase and stearoyl-CoA desaturase 1: Role of MEK/ERK1/2 pathway.

AIM: To investigate the effect of MEK/ERK1/2 pathway on peroxisome proliferator-activated receptors (PPARγ) agonist-induced alterations in Δ6-desaturase (Δ6D) and stearoyl-CoA desaturase 1 (SCD1) in hepatocellular carcinoma cell line HepG2. METHODS: HepG2 cells cultured in RPMI-1640 were exposed to the commonly used ERK1/2 pathway inhibitor PD98059 and PPARγ agonist, pioglitazone. Total RNA was isolated and reverse transcribed from treated cells. Changes in gene expression and metabolites ratio, as activity index for Δ6D and SCD1, were then determined using reverse transcription-polymerase chain reaction and gas liquid chromatography, respectively. RESULTS: The expression of both Δ6D (P = 0.03) and SCD1 (P = 0.01) increased following PD98059 treatment, with a higher impact on SCD1 (24.5% vs 62.5%). Although pioglitazone increased the mRNA level (1.47 ± 0.10 vs 0.88 ± 0.02, P = 0.006) and activity index (1.40 ± 0.07 vs 0.79 ± 0.11, P < 0.001) of Δ6D, no such changes have been observed for SCD1 activity index in pioglitazone-treated cells. SCD1 gene expression (+26.4%, P = 0.041) and activity index (+52.8%, P = 0.035) were significantly increased by MEK inhibition in the presence of pioglitazone, as compared with pioglitazone alone and control cells. However, the response of Δ6D expression and activity index to pioglitazone was unaffected by incubation with PD98059. CONCLUSION: PPARγ and ERK1/2 signaling pathway affect differentially and may have inhibitory crosstalk effects on the genes expression of ∆6D and SCD1, and subsequently on their enzymatic activities.

Fatty acid composition of human follicular fluid phospholipids and fertilization rate in assisted reproductive techniques.

BACKGROUND: Fatty acids are known to be critically important in multiple biological functions. Phospholipid fatty acids of follicular fluid, an important microenvironment for the development of oocytes, may contribute to the women's fertility and the efficacy of assisted reproduction techniques. The aim of this study was to investigate the effect of fatty acid composition of follicular fluid phospholipids on women undergoing assisted reproductive techniques. METHODS: Follicular fluid samples were obtained from 100 patients, referred to Tabriz Alzahra Hospital. Seventy-nine subjects underwent in vitro fertilization (IVF) and the remaining 21 underwent intracytoplasmic sperm injection (ICSI). Total lipid of follicular fluid was extracted and fatty acids were analyzed by gas-liquid chromatography. RESULTS: Saturated fatty acids (SFA, P = 0.002) and the ratio of SFA to polyunsaturated fatty acids (P = 0.001) were correlated negatively with a number of mature oocytes after age adjustment. Linoleic acid (P = 0.006) was positively correlated, while the level of arachidonic acid was negatively correlated with fertility percentage after adjustment for body mass index, sperm count, sperm motility. CONCLUSION: Since phospholipids are one of the major components of lipid metabolism, the results of this study highlight the importance of this component in follicular fluid lipid metabolism. Consequently, it is proposed as an index in determination of the rate of success in assisted reproductive techniques such as IVF/ICSI.

Correlation between PPARγ protein expression level in granulosa cells and pregnancy rate in IVF program.

BACKGROUND: Peroxisome proliferative-activated receptors (PPARs) are nuclear receptors that involved in cellular lipid metabolism and differentiation. The subtype γ of the PPAR family (PPARγ) plays important roles in physiologic functions of ovaries. OBJECTIVE: To determine correlation between PPARγ protein level in granulosa cells and pregnancy rate in women undergoing in-vitro fertilization (IVF) treatment. MATERIALS AND METHODS: In this cross-sectional study, twenty-five samples of granulosa cells were collected from women referred to an IVF treatment center. PPARγ protein expression level in granulosa cells was determined in comparison with ß-actin level as control gene with Western blot test. Laboratory pregnancy was determined by a rise in blood ß-hCG level fourteen days after embryo transfer. Correlation analyses were used to test for associations between the oocytes and pregnancy occurrence as outcome variables and PPARγ protein expression level. RESULTS: Correlation analysis indicated that there was no significant relationship between granulosa cells PPARγ protein level with IVF parameters including number of matured oocytes and the ratio of fertilized to matured oocytes. Comparison of granulosa cells PPARγ protein level with positive and negative laboratory pregnancy revealed also no significant relationship. CONCLUSION: According to the results of this study, PPARγ protein level in granulosa cells could not be directly correlated to the success rate of IVF.

Inhibition of MEK/ERK1/2 Signaling Affects the Fatty Acid Composition of HepG2 Human Hepatic Cell Line.

INTRODUCTION: The extracellular signal-regulated kinase (ERK) mitogen-activated protein kinase pathway, also known as the MEK/ERK1/2 kinase cascade, has recently been implicated in the regulation of lipid metabolism and fatty liver disease. However, its functional effect on cellular fatty acid composition is unknown. Herein, we examined the effect of a pharmacological inhibitor of MEK, the upstream kinase activator of ERK1/2, on fatty acid composition of hepatocellular carcinoma cell line HepG2. METHODS: HepG2 cells cultured in RPMI-1640 were exposed to the commonly used ERK1/2 pathway inhibitor PD98059 and were investigated with respect to fatty acid composition by gas-liquid chromatography. RESULTS: Exposure of cells to the ERK1/2 pathway inhibitor induced an increase in monounsaturated fatty acids and the fatty acid desaturation index and a decrease in polyunsaturated fatty acid content. Specifically, we showed a significant increase of oleic acid (18:1n-9; +29%, P=0.003) and arachidonic acid (20:4n-6)/linoleic acid (18:2n-6) ratio (3.5-fold; P<0.001) in HepG2 cells. CONCLUSION: Cellular fatty acid composition of HepG2 cells appeared to be differentially regulated by ERK1/2 pathway, thus suggesting related metabolic pathways as potential mediators of the effects of ERK1/2 signaling on hepatic fatty acid composition.

Interleukin-1 beta, interleukin-6 and TGF-beta in follicular tissue of impacted third molars.

The clinical evaluation and management of impacted third molars remain challenging. The aim of the present study was to investigate possible associations between follicular tissue cytokines and radiographic manifestations of impacted third molar. The population included 72 patients who underwent surgical extraction of impacted third molars. All these patients underwent a preliminary panoramic radiograph. Levels of interleukin-1 beta (IL-1ß), interleukin-6 (IL-6) and transforming growth factor beta (TGF-ß) in tissue extracts were determined using ELISA. There were no significant differences between bony and tissue impaction as regards IL-1ß, IL-6 and TGF-ß levels. Moreover, the same results were obtained as far as the amount of pericoronal space and the presence or absence of a history of pericoronitis are concerned. These results suggest that radiographic findings or a history of pericoronitis are not associated with levels of expression of pro-inflammatory cytokines in patients undergoing surgical removal of impacted third molars. However, further studies are needed to address the possibility of variability during disease progression.

Two common polymorphisms in the peroxisome proliferator-activated receptor γ gene may improve fertilization in IVF.

Genetic factors play an important role in women's fertility and embryonic development and may also contribute to the efficacy of assisted reproduction techniques. The aim of this study was to investigate the effect of His447His and Pro12Ala peroxisome proliferator-activated receptor γ (PPARγ) gene polymorphisms on oocytes and fertilization in women undergoing IVF. Follicular fluid and blood samples were obtained from 98 IVF patients referred to Tabriz Alzahra Hospital. Samples were analysed for fatty acid content by gas-liquid chromatography and for polymorphisms of the PPARγ gene using polymerase chain reaction-restriction fragment length polymorphism-based methods. Multiple regression analyses were used to test the independence of associations between the number of mature and fertilized oocytes as outcome variables and the polymorphisms of PPARγ gene. For both polymorphisms, fertilization ratio was significantly (P<0.05) higher in carriers of the rare alleles than homozygous wild-type genotypes. The associations of His447His (P=0.003) and Pro12Ala (P=0.015) polymorphisms remained statistically significant in the multiple regression analyses. This study suggests that the two common gene polymorphisms of PPARγ may improve fertilization in vitro and, thus possibly, female fertility.

Fatty acid composition of epicardial and subcutaneous human adipose tissue.

BACKGROUND: Epicardial adipose tissue has special properties that distinguish it from the more widely studied depots of adipose tissue. OBJECTIVES: We undertook this study to investigate regional differences between epicardial and subcutaneous adipose tissue fat composition, as well as associations between these measures and metabolic variables. METHODS: Epicardial and subcutaneous peripheral adipose tissue were collected during coronary artery bypass grafting from 42 patients (ages 37-65) with coronary artery disease (CAD). The fatty acid composition of adipose tissue was determined by gas liquid chromatography (GLC). RESULTS: The saturated fatty acids, including myristic acid (14:0), palmitic acid (16:0), and stearic acid (18:0), were higher, and the unsaturated fatty acids, including palmitoleic acid (16:1n-7), oleic acid (18:1n-9), linoleic acid (18:2n-6), and linolenic acid (18:3n-3), were lower than the subcutaneous adipose tissue. The presence of hypertension was positively correlated with the 16:1n-7 (r = 0.407, P = 0.032) and 18:1n-11(r = 0.370, P = 0.027), and negatively correlated with 18:1n-9 (r = -0.367, P = 0.036) and 18:2n-6 (r = -0.446, P = 0.006) contents of epicardial adipose tissue after adjustment for body mass index. CONCLUSIONS: Regional differences may be observed in fatty acid composition, suggesting a depot specific impact of stored fatty acids on adipocyte function and metabolism.