Improvement of liver cell therapy in rats by dietary stearic acid

Background: Stearic acid is known as a potent anti-inflammatory lipid. This fatty acid has profound and diverse effects on liver metabolism. The aim of this study was to investigate the effect of stearic acid on markers of hepatocyte transplantation in rats with acetaminophen [APAP]-induced liver damage

Methods: Wistar rats were randomly assigned to 10-day treatment. Stearic acid was administered to the rats with APAP-induced liver damage. The isolated liver cells were infused intraperitoneally into rats. Blood samples were obtained to evaluate the changes in the serum liver enzymes, including activities of aspartate aminotransferase [AST], alanine aminotransferase [ALT] and alkaline phosphatase [ALP] and the level of serum albumin. To assess the engraftment of infused hepatocytes, rats were euthanized, and the liver DNA was used for PCR using sex-determining region Y [SRY] primers

Results: The levels of AST, ALT and ALP in the serum of rats with APAP-induced liver INJURY were significantly increased and returned to the levels in control group by day six. The APAP-induced decrease in albumin was significantly improved in rats through cell therapy, when compared with that in the APAP-alone treated rats. SRY PCR analysis showed the presence of the transplanted cells in the liver of transplanted rats

Conclusion: Stearic acid-rich diet in combination with cell therapy accelerates the recovering of hepatic dysfunction in a rat model of liver injury

Correlation of serum lipoprotein ratios with insulin resistance in infertile women with polycystic ovarian syndrome: a case control study

Background: Dyslipidemia and insulin resistance [IR], occurring in most infertile women with polycystic ovarian syndrome [PCOS], increase the risk of cardiovascular disease [CVD] and type 2 diabetes. This study aimed to assess the relationships between lipopro-tein ratios and IR in PCOS women

Materials and Methods: Thirty six infertile women with PCOS selected based on Andro-gen Excess Society [AES] criteria and 29 healthy women matched for age were recruited to this case-control study. After physical measurements, fasting serum glucose [Glu], insulin and lipid profile levels [triglycerides [TGs], total cholesterol [TC], low-density lipoprotein-cholesterol [LDL-C] and high-density lipoprotein-cholesterol [HDL-C]] were measured, while lipoprotein ratios [TC/HDL-C, LDL-C/HDL-C, TG/HDL-C] were calculated. IR was also calculated using homeostasis model assessment [HOMA]-IR. The optimal cutoffs of lipoprotein ratios in relation to HOMA-IR were calculated based on the Receiver Operating Characteristics [ROC] curve analysis using the area under curve [AUC]

Results: Waist circumference [WC], insulin levels, HOMA-IR, TG levels, and all lipoprotein ratios were significantly higher, while HDL-C was lower in PCOS group as compared to healthy controls. All lipoprotein ratios, TG levels, and WC are significantly correlated with insulin levels and HOMA-IR. Among lipoprotein ratios, the highest AUC of the ROC belonged to TG/HDL-C ratio with sensitivity of 63.6% and specificity of 84.4% [TG/HDL-O3.19] as a marker of IR in infertile PCOS women

Conclusion: Lipoprotein ratios, particularly TG/HDL-C, are directly correlated with insulin levels and can be used as a marker of IR [HOMA-IR] in infertile PCOS patients

Influence of omega-3 fatty acid eicosapentaenoic acid on IGF-1 and COX-2 gene expression in granulosa cells of PCOS women

The omega-3 [omega-3] fatty acid eicosapentaenoic acid [EPA] is currently used in the clinic as a nutritional supplement to improve infertility, particularly in women with polycystic ovarian syndrome [PCOS]. The present study was designed to investigate the effect of EPA on insulin-like growth factor 1 [IGF-1] and cyclooxygenase 2 [COX-2] gene expression in primary cultured granulosa cells from patients undergoing in vitro fertilization [IVF], and also to compare this effect with those in granulosa cells of PCOS patients. In this experimental study, human granulosa cells were isolated from follicular fluid of normal and PCOS women undergoing IVF by hyaluronidase digestions, followed by Percoll gradient centrifugation. Cells were cultured in vitro, exposed to a range of concentrations of the EPA [25-100 micro M] for 24 hr, and investigated with respect to COX-2 and IGF-1 gene expression by real time-PCR. In both groups, all doses of the EPA significantly induced IGF-1 mRNA gene expression compared to the untreated control. High doses of EPA in the presence of recombinant [r] FSH produced a stimulatory effect on IGF-1 and a suppressive effect [p=0.01] on the COX-2 gene expression, which were more pronounced in granulosa cells from PCOS patients. EPA affect diversely the gene expression of IGF-1 and COX-2 in granulosa cells, which were more pronounced in PCOS compared to control. These findings represent the possible underlying molecular mechanisms for the positive impact of the omega-3 fatty acids on reproduction, especially in patients with PCOS

763C>G polymorphism of the secretory PLA2IIa gene is associated with endometriosis in Iranian women

Endometriosis is a chronic gynecological disease resulting from complex interactions between genetic, hormonal, environmental and oxidative stress and intrinsic inflammatory components. The aim of this study was to investigate the potential association of the 763C>G polymorphism in the secretory phospholipase A2 group IIa gene [PLA2G2A] with the risk of endometriosis in Iranian women. Ninety seven patients with endometriosis along with 107 women who were negative for endometriosis after laparoscopy and laparatomy, and served as the control group, were enrolled for this cross-sectional study. Samples were genotyped using the polymerase chain reaction-restriction fragment length polymorphism method. Multivariate analysis was used to examine the association between the risk of endometriosis and the 763C>G polymorphism of PLA2G2A. Genotype distributions of PLA2G2A were significantly different between patients and the controls [p<0.001, OR=0.22, 95% CI=0.21-0.39]. Correlation analysis showed that there was a significant association between the normal homozygous genotype and susceptibility to endometriosis [p<0.001]. The present study suggests that the 763C>G polymorphism of PLA2G2A plays an important role as an independent factor in the risk of endometriosis in Iranian women

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

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 microM), 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 Delta9 desaturation, modulate other reactions in de novo fatty acid synthesis and lipogenesis, and subsequently affect the overall survival and progression of breast cancer.

High omega-3: omega-6 fatty acids ratio increases fatty acid binding protein 4 and extracellular secretory phospholipase A2IIa in human ectopic endometrial cells

Endometriosis, a common chronic inflammatory disorder, is defined by the atypical growth of endometrium- like tissue outside of the uterus. Secretory phospholipase A2 group Ha [sPLA2-IIa] and fatty acid binding protein4 [FABP4] play several important roles in the inflammatory diseases, Due to reported potential anti-inflammatory effects of omega-3 and omega-6 fatty acids, the purpose of the present study was to investigate the effects of omega-3 and omega-6 polyunsaturated fatty acids [PUFAs] on fatty acid binding protein 4 and extracellular secretory phospholipase A2IIa in cultured endometrial cells. Ectopic and eutopic endometrial tissues obtained from 15 women were snap frozen. After thawing and tissue digestion, primary mixed stromal f and endometrial epithelial cell culture was performed for 8 days in culture mediums supplemented with normal and high ratios of omega-3 and omega-6 PUFA. sPLA2-IIa in the J culture medium and FABP4 level was determined using enzyme immuno assay [EIA] technique. Within ectopic endometrial cells group, the level of cellular FABP4 and extracellular sPLA2-IIa were remarkably increased under high omega-3 PUFA exposure compared with control condition [p=0.014 and p=0.04 respectively]. Omega-3 PUFAs may increase the level of cellular FABP4 and extracellular sPLA2-IIa in ectopic endometrial cells, since sPLAIIa and FABP4 may affect endometriosis via several mechanisms, more relevant studies are encouraged to know the potential effect of increased cellular FABP4 and extracellular sPLA2-IIa on endometriosis

Paraoxonase 3 activity and the ratio of antioxidant to peroxidation in the follicular fluid of infertile women

Paraoxonase-3 [PON3], as a high density lipoprotein [HDL]-associated lactonase, is capable of preventing the oxidative modification of low density lipoprotein [LDL]. PON3 activity in follicular fluid [FF] is three times more than its activity in serum. However, the detailed role of PON3 in women's fertility remains unknown. The aim of this study was to investigate the correlation between PON3 activity in the FF of women undergoing assisted reproductive technique [ART], in vitro fertilization [IVF], or intra-cytoplasmic sperm injection [ICSI]. This cross-sectional study consisted of 50 women from couples with male factor infertility [MFI] or with female factor infertility [FFI]. The FF samples were obtained during the ART intervention. PON3 activity, HDL cholesterol [HDL C], total antioxidant status [TAS] and the level of malondialdehyde [MDA] were determined. The morphology of the embryo was determined using embryo cell number [ECN] and embryo fragmentation score [EFS]. In addition, fertilization rate [FR] was used an oocyte fertilization index. Of 50 women, 20 women belonged to FFI group and the remaining 30 women belonged to MFI group. PON3 activity in FF of women in FFI group was significantly lower [p<0.05] in comparison with corresponding value in MFI group. The value of PON3 activity/MDA in the FFI group was lower than that in MFI group. Moreover, MDA level in the FF of FFI group was significantly higher [p<0.05] than its concentration in MFI group. Meanwhile, no significant difference was found in HDL-C concentration and TAS of both groups. No significant correlation was observed between the ECN and FF biochemical parameters. There was also a negative correlation between FR and MDA [r=-0.42, p=0.02], whereas a positive relation between FR with PON3 activity [r=0.59, p=0.004], HDL-C [r=0.35, p=0.04] and PON3/MDA [r=0.59, p=0.001]. Conclusion: According to the results of this study, PON3 activity level as a key component of antioxidant system in FF may directly be associated with the success rate of ART and fertilization rate in women

Correlation between PPAR gamma protein expression level in granulosa cells and pregnancy rate in IVF program

Peroxisome proliferative-activated receptors [PPARs] are nuclear receptors that involved in cellular lipid metabolism and differentiation. The subtype gamma of the PPAR family [PPAR gamma] plays important roles in physiologic functions of ovaries. To determine correlation between PPAR gamma protein level in granulosa cells and pregnancy rate in women undergoing in-vitro fertilization [IVF] treatment. In this cross-sectional study, twenty-five samples of granulosa cells were collected from women referred to an IVF treatment center. PPAR gamma protein expression level in granulosa cells was determined in comparison with beta -actin level as control gene with Western blot test. Laboratory pregnancy was determined by a rise in blood beta -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 PPAR gamma protein expression level. Correlation analysis indicated that there was no significant relationship between granulosa cells PPAR gamma protein level with IVF parameters including number of matured oocytes and the ratio of fertilized to matured oocytes. Comparison of granulosa cells PPAR gamma protein level with positive and negative laboratory pregnancy revealed also no significant relationship. According to the results of this study, PPAR gamma protein level in granulosa cells could not be directly correlated to the success rate of IVF

Evaluation of the relationship between endometriosis and omega-3 and omega-6 polyunsaturated fatty acids

Endometriosis is a common chronic inflammation causing major problems including infertility. The role of omega-3 and omega-6 fatty acids as their potential anti-inflammatory effects in endometriosis needs to be further explored. The objective of this study was to compare serum phospholipid fatty acid profile in endometriosis patients with controls, and to explore the correlation of this profile with the severity of the disease. Sixty-four endometriosis patients and 74 control women, in reproductive age, participated in this study. Among the endometriosis patients, 19 cases were in stage I, 27 cases in stage II, 8 cases in stage III, and 10 cases in stage IV. Each patient underwent laparoscopy. Before surgery, 5 ml of blood was obtained. After extraction of the total lipids, serum total phospholipid fraction was isolated by thin layer chromatography. Fatty acid composition of the phospholipid fraction was determined by gas chromatography and the resulted profile was compared in endometriosis patients and controls. The profile was also compared in the endometriosis group based on the severity of disease. Stearic acid was significantly lower in the endometriosis group as compared to controls [P= 0.030]. No other fatty acid compositions were significantly different between patients and controls. Serum ratio of eicosapentaenoic acid [EPA] to arachidonic acid [AA] was in reasonable correlation with the severity of endometriosis [r = 0.34, P = 0.006]. According to these findings, levels of fatty acids in serum total phospholipids seem not to be a marker for endometriosis, but the EPA to AA ratio was a relevant factor indicating severity of illness

Effect of polyunsaturated fatty acids on secretory phospholipase A2 type IIa in ectopic endometrial cells

Endometriosis is a common chronic inflammation which leads to infertility and chronic pelvic pain in affected women. Secretory phospholipase A2 type IIa [sPLA2IIa] is an acute phase reactant that is markedly increased in inflammatory disorders. To assess the effects of omega-3 and omega-6 polyunsaturated fatty acids [PUFAs] administration in endometrial cells culture on sPLA2IIa level and cell survival comparing homolog ectopic versus eutopic endometrial cells from endometriosis patients. In this experimental study, ectopic and eutopic endometrial tissue samples obtained from 15 endometriosis patients were immediately frozen. After thawing and tissue digestion, mixed stromal and endometrial gland cells were cultured for 8 days in three different culture media; balanced omega-3/omega-6, high omega-3 and high omega-6 PUFAs ratio. Cell survival was measured using 2, 3-bis [2-methoxy-4-nitro-5-sulfophenyl]-5-[phenylamino] carbonyl-2H- tetrazolium hydroxide [XTT] method and sPLA2IIa level assessed with ELISA technique. The sPLA2IIa level was significantly higher in the ectopic endometrial cell culture compared to the eutopic group for each of the three matched treatments [balanced, high omega-3 and high omega-6]. Also the sPLA2IIa level in the ectopic endometrial cell group was remarkably increased by each of the three PUFAs treatments compared to control condition [p<0.05, p<0.01, p<0.05 respectively]. Cell survival in the eutopic group was significantly decreased by high omega-6 culturing compared to control medium [p<0.05]. The increase in sPLA2IIa level in ectopic endometrial cells by fatty acid treatments [especially high omega-3], strengthens the hypothesis that PUFAs stimulate secretion of cytokines leading to increased sPLA2IIa level

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

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. 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. 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. 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 the level of cholesteryl ester transfer protein in follicular fluid with fertilization rates in IVF/ ICSI cycles

Follicular fluid [FF] plays an important role in oocytes and embryo development, which may contribute to IVF/ICSI success rate. The aim of this study was to investigate the correlation between cholesteryl ester transfer protein [CETP] level in FF and the success rate of IVF/ICSI. In a cross-sectional study, FF samples, FF samples were obtained from 100 patients referred to Tabriz Alzahra Hospital. Seventy-nine subjects underwent IVF and the remaining 21 underwent ICSI. The levels of high-density lipoprotein cholesterol [HDL-C], apolipoprotein A-I and CETP were measured using enzymatic, turbidometric and ELISA methods respectively. Analysis of the subgroups with different levels of CETP showed a significant lower level of CETP in the subgroup with the lowest number of mature oocytes [p<0.05]. The level of CETP was also considerably lower [18%, p=0.05] in subjects with<50% oocytes fertilization ratio than subjects with >70% of this ratio. While no association was found for pregnancy, the amount of CETP in FF was associated positively to the maturity and the percentage of oocyte fertilization

I405V and -629C/A polymorphisms of the cholesteryl ester transfer protein gene in patients with coronary artery disease

Cholesteryl ester transfer protein [CETP] plays a main role in high-density lipoprotein metabolism. CETP gene possesses several single nucleotide polymorphisms which have been associated with plasma high-density lipoprotein cholesterol [HDL-C] concentrations. The aim of this study was to determine the association of CETP -629C/A and I405V polymorphisms with coronary artery disease [CAD] in Iranian population. The presence of two CETP gene polymorphisms -629C/A and I405V were studied in 187 unrelated CAD cases and 136 controls. All the samples were clinically examined and lipid profile was estimated. Genotyping was performed using polymerase chain reaction/restriction fragment length polymorphism method. The frequency of-629C/A and I405V allelic variants were found to be 0.732 and 0.366 in cases and 0.658 and 0.348 in controls, respectively. The frequency of A allele of -629C/A polymorphism in cases was significantly higher than that of controls. HDL-C in AA genotype was higher than CA and CC genotypes in controls. No significant effect of II, IV and VV genotypes was found in lipid profiles. No significant association was found between CETP I405V polymorphism and increased risk of CAD in Azeri population studied. AA genotype of -629C/A increased HDL but the risk of CAD in this genotype might be higher than CC genotype