Correlation between follicular fluid 25-OH vitamin D and assisted reproductive outcomes

Vitamin D in complex with its receptor by regulating gene expression, endometrium immune response and stimulation of endometrium decidualization can be involved in implantation. So, it seems that the amount of vitamin D in follicular fluids [FF] may have an association with ART success. First, we intended to investigate the possible association between levels of follicular fluids 25-OH vitamin D with assisted reproductive outcomes. Second, we examined relationship between 25-OH vitamin D levels with number and quality of oocytes. In a prospective study, 80 infertile female candidates for IVF/ICSI were enrolled. Blood samples [on the day of human chorionic gonadotropin administration] and follicular fluids were taken, and then levels of serum estradiol and follicular fluids 25-OH vitamin D were measured. Also clinical characteristics of patients [duration of infertility, causes of infertility, menstrual status], number and quality of oocytes, number of fertilized oocytes, estradiol levels, and clinical pregnancy were evaluated. Concentration of FF 25-OH vitamin D in pregnant women was significantly higher than non-pregnant women [p=0.007] but there were no significant differences in age, body mass index [BMI], duration of infertility, menstrual status, number of oocytes, oocytes quality, number of fertilized oocytes, and serum estradiol levels between the two groups. Statistically positive correlation was found between 25-OH vitamin D levels with patient age and implantation rate [r=0.264, p=0.018 and r=0.301, p-0.007 respectively]. The obtained results suggest that vitamin D without affecting the number and quality of oocytes can independently improve implantation rate and 1VF outcome

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

Altered of microRNA expression level in oligospermic patients

MicroRNA [miRNA] is small endogenous, single strand RNA molecules that regulate gene expression at post-transeriptional level through several mechanisms to affect key cellular event including male germ cells differentiation, proliferation, development and apoptosis. Mutation and/or aberrant expression of miRNAs have been associated with progression of various disorders, including infertility. The purpose of this research was to study the estrogen receptor beta [ERbeta], hsa-mir-21 and, hsa-mir-22 expression level in oligospermic infertile and control fertile men and correlation between them. In this study, the change in mir-21, mir-22 expression and their common target gene [ERbeta] expression levels were evaluated in oligospermic infertile men [n= 43] compared with 43 age matched healthy control by Real-Time PCR methods. Expression analysis by qRT-PCR test on miRNA have identified that mir-21, mir-22 levels were significantly higher than those in normal controls [p<0.0001] and ER beta expression level significantly decreased in comparison with the normal group [p<0.0001]. Our study showed that mir-21 and mir-22 are indirectly involved in spermatogenesis by regulating of the estrogen receptor and might have a diagnostic and prognostic value in men infertility

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

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 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

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