scholarly journals Predict Value of Relative Telomere Length, TERT, and Follicular Fluid Anti-mullerian Hormone Levels for IVF Outcome in Infertile Women

2021 ◽  
Author(s):  
Yan Hao ◽  
Mingrong Lv ◽  
Jing Peng ◽  
Zhihua Zhang ◽  
Zhiguo Zhang ◽  
...  
Keyword(s):  
Telomere Length ◽  
Granulosa Cells ◽  
Follicular Fluid ◽  
Significant Relationship ◽  
Clinical Pregnancy ◽  
Human Beings ◽  
Expression Levels ◽  
Infertile Women ◽  
Relative Telomere Length ◽  
Tert Expression

Abstract Background: Telomere attrition has been shown to play a critical role in the reproductive aging process in human beings. Telomere length (TL) is normally regulated by telomerase enzyme. Telomerase reverse transcriptase (TERT) is the main component of the telomerase. Anti-Mullerian hormone (AMH), a member of the transforming growth factor superfamily, is derived from the granulosa cells of early developing pre-antral and antral follicles. The aim of this study was to evaluate the associations between relative telomere length (RTL), TERT expression of granulosa cells (GC), follicular fluid (FF) AMH levels and ovarian/embryonic performance in infertile women at different age. Moreover, whether they acting as predictors for probability of clinical pregnancy were also assessed. Method: A total of 160 women underwent their first fresh cycle of in vitro fertilization (IVF) or intracytoplasmic sperm injection (ICSI) were included in our study as follows: 100 women were enrolled for RTL measurement and 60 women were enrolled for TERT measurement. All these 160 women underwent FF AMH measurement. Correlations between RTL，TERT expression, FF AMH levels and age, ovarian/embryonic performance and probability of clinical pregnancy were assessed.Results: There was a statistically significant relationship between the expression levels of TERT, RTL, FF AMH levels and patient age(r = −0.20, P = 0.04; r=0.30, P=0.02; r=-0.191, P=0.003, respectively). Relationships between the expression levels of TERT, FF AMH levels and oocytes yield were significant (P<0.001; P<0.05, respectively). However, no statistically correlation was observed between the RTL of GC samples and oocytes yield. All these three biomarkers had no correlation with blastocyst formation rate. There was significant relationship between FF AMH levels and probability of clinical pregnancy in patients older than 35 years (OR=1.284, 95%CI=1.031-1.599, P=0.026). Conclusion: RTL, relative TERT expression in GC and AMH levels in follicular fluid are age-related, but all of them fail to predict embryonic outcomes. Relative TERT expression and FF AMH levels appear to be more reliable for prediction of ovarian response than RTL. FF AMH is also a good predictor for probability of clinical pregnancy in advanced women.

P–565 Does length really matter? The impact of lifestyle on granulosa cell telomeres length and IVF outcome

2021 ◽  
Vol 36 (Supplement_1) ◽  
Author(s):  
O Limonad ◽  
S Hantisteanu ◽  
S Meise. Sharon ◽  
N Haggiag ◽  
D Estrada ◽  
...  
Keyword(s):  
Treatment Outcome ◽  
Telomere Length ◽  
Pregnancy Rate ◽  
Quantitative Pcr ◽  
Granulosa Cells ◽  
Follicular Fluid ◽  
Relative Length ◽  
Oocyte Retrieval ◽  
Relative Telomere Length ◽  
The Impact

Abstract Study question Is the relative telomere length in granulosa cells associated with an abnormal metabolic profile of IVF patients and treatment outcome? Summary answer Longer telomeres and higher pregnancy rates have been observed in women with higher follicular fluid glucose levels. What is known already Telomeres may serve as a biomarker of cell senescence; their length varies depending on two factors: genetic predisposition and shortening processes related to cell division. In several studies, telomere shortening in granulosa cells has been shown to be correlated with ovarian aging. However, these findings are still limited in applicability, and more research is required to define the factors involved. Study design, size, duration Women undergoing IVF participated in a prospective cohort study between 2018 and 2019. Peripheral blood was obtained on the day of oocyte retrieval. Pooled samples of follicular fluid were collected for a comprehensive telomere length analysis in granulosa and cumulus cells by quantitative PCR (qPCR). Participants/materials, setting, methods DNA was extracted from granulosa cells and assessed for relative telomere length by monochrome multiplex quantitative PCR (qPCR). Telomere length was then analyzed relative to a single copy gene (36B4) to evaluate possible metabolic profile’s impact on telomere length and treatment outcome. Hormonal profile, chemistry, and inflammation factors were analyzed in the follicular fluid and serum. Main results and the role of chance Out of forty-nine women recruited for the study, forty-one cases were eligible for a comprehensive analysis. Follicular fluid CRP and triglyceride levels in BMI-based analysis were significantly lower in women with BMI &lt;25 compared to those with BMI &gt;30 (0.5 (0.2–1.5) vs. 6.6 (3.6–10.6); p &lt; 0.0001 and 21.04 ± 8.04 vs. 28.18 ± 8.97; p = 0.011, respectively). Interestingly, a significant correlation between the relative lengths of telomeres and pregnancy rate was observed (p &lt; 0.001), with a higher pregnancy rate in women with longer telomeres (88%) than in women with shorter telomeres (38%). Relatively longer telomeres (0.96 (0.94–0.99) vs. 0.91 (0.82–0.95), p = 0.02) and higher levels of follicular fluid glucose (63±11.12 vs. 50.67±15.69, p = 0.006) were observed in those who conceived. No statistically significant differences were detected in the levels of the other hormones measured in the blood or follicular fluid on the day of oocyte retrieval or other clinical parameters, including follicle size and number, embryo quality, and the number of OHSS cases. Limitations, reasons for caution As a preliminary study, there were few participants. Further implications can be reached by increasing the sample size, but telomerase activity evaluation is required to confirm telomere length measurements without question. Whether short telomeres associate with infertility as a cause or consequence of BMI warrant more research. Wider implications of the findings: The relative length of telomeres in granulosa cells at the time of oocyte retrieval may serve as a predictive biomarker for oocyte competence and subsequent pregnancy. Moreover, healthy lifestyle behavior is recommended. Trial registration number Not applicable

scholarly journals Expression of mRNAs for Pro-and Anti-Apoptotic Factors in Granulosa Cells and Follicular Fluid of Women Undergoing in Vitro Fertilization. A Pilot Study

2021 ◽  
Author(s):  
Jozsef Bodis ◽  
Endre Sulyok ◽  
Akos Varnagy ◽  
Viktória Prémusz ◽  
Krisztina Godony ◽  
...  
Keyword(s):  
Gene Expression ◽  
In Vitro Fertilization ◽  
Mrna Expression ◽  
Granulosa Cells ◽  
Follicular Fluid ◽  
Vitro Fertilization ◽  
Expression Levels ◽  
The Impact ◽  
Apoptotic Factors

Abstract BackgroundThis observational clinical study evaluated the expression levels and predictive values of some apoptosis-related genes in granulosa cells (GCs) and follicular fluid (FF) of women undergoing in vitro fertilization (IVF).Methods GCs and FF were obtained at oocyte retrieval from 31 consecutive patients with heterogeneous infertility diagnosis (age: 34.3±5.8 years, body mass index: 24.02±3.12 kg/m2, duration of infertility: 4.2±2.1 years). mRNA expression of pro-apoptotic (BAX, CASP3, CASP8) and anti-apoptotic (BCL2, AMH, AMHR, FSHR, LHR, CYP19A1) factors was determined by quantitative RT-PCR using ROCHE LightCycler 480. Results No significant difference in GC or FF mRNA expression of pro- and anti-apoptotic factors could be demonstrated between IVF patients with (9 patients) or without (22 patients) clinical pregnancy. Each transcript investigated was detected in FF, but their levels were markedly reduced and independent of those in GCs. The number of retrieved oocytes was positively associated with GC AMHR (r=0.393, p=0.029), but the day of embryo transfer was negatively associated with GC LHR (r=-0.414, p=0.020) and GC FSHR transcripts (r=-0.535, p=0.002). When pregnancy positive group was analysed separately the impact of apoptosis- related gene expressions on some selected measures of IVF success could be observed. Strong positive relationship was found between gene expression levels of pro- and anti-apoptotic factors in GCs.ConclusionOur study provides only marginal evidences for the apoptosis dependence of IVF outcome and suggests that the apoptosis process induces adaptive increases of the anti-apoptotic gene expression to attenuate apoptosis and to protect cell survival.

scholarly journals Evaluation of Presepsin as a Predictive Marker in Women Undergoing ICSI

2020 ◽  
Vol 1 (6) ◽  
Author(s):  
Bushra Rasheed Al-Azawea ◽  
Hayder A. L. Mossa ◽  
Liqaa R. Altamimi ◽  
Lubna Amer Al-Anbary
Keyword(s):  
Follicular Fluid ◽  
Intracytoplasmic Sperm Injection ◽  
Ovarian Stimulation ◽  
Predictive Marker ◽  
Clinical Findings ◽  
Clinical Pregnancy ◽  
Infertile Women ◽  
Poor Predictor ◽  
Significant Difference ◽  
Antagonist Protocol

Purpose: The purpose of this study was to determine the ability of using Presepsin as a biomarker to evaluate the clinical pregnancy rate in a cohort of couples undergoing ICSI in a sample of Iraqi population. Patients and Methods: Sixty infertile women selected for undergoing intracytoplasmic sperm injection, the patient’s ovarian stimulation were by antagonist protocol accordingly to their clinical findings. Results: The results of the study shows there was no significant difference in the serum Presepsin between pregnant and non-pregnant ladies, 421.57 (1534.65) versus 878.64 (1322.23), respectively (P = 0.875), Furthermore, there was no significant difference in follicular fluid Presepsin between pregnant and non-pregnant ladies, 1286.73 (1232.00) versus 0.00 484.15 (1467.00), respectively (P = 0.296). Conclusions :We conclude that serum or follicular fluid Presepsin as a biomarker is a poor predictor of fertility outcome with no significant difference between pregnant and non-pregnant ladies

scholarly journals TRIB3 Regulates FSHR Expression in Human Granulosa Cells Under High Levels of Free Fatty Acids

2021 ◽  
Author(s):  
Nan Wang ◽  
Chenchen Si ◽  
Lan Xia ◽  
Xian Wu ◽  
Sheng Zhao ◽  
...  
Keyword(s):  
Fatty Acids ◽  
Free Fatty Acids ◽  
Signaling Pathway ◽  
Oocyte Maturation ◽  
Granulosa Cells ◽  
Follicular Fluid ◽  
Glycogen Synthase ◽  
Obese Women ◽  
Expression Levels

Abstract Background: Granulosa cells (GCs) in cumulus oophorus highly express follicle stimulating hormone receptor (FSHR), which is the most important mediator of both estradiol synthesis and oocyte maturation. Obese women have elevated free fatty acids (FFAs) levels in their follicular fluids and decreased FSHR expression in GCs, which is related to an altered protein kinase B/glycogen synthase kinase 3β (Akt/GSK3β) signaling pathway. Such FFA increases accompany 3-fold rises in pseudokinase 3 (TRIB3) expression and reduce the Akt phosphorylation status in both the human liver and in insulinoma cell lines. Therefore, in a high FFA environment, we determined if TRIB3 mediates regulation of FSHR via the Akt/GSK3β signaling pathway in human GCs. Methods: GCs from women undergoing in vitro fertilization were collected and designated as high and low FFAs cohorts based on their follicular fluid FFA content. GCs with low FFA levels and a human granulosa-like tumor (KGN) cell line were exposed to palmitic acid (PA), which is a dominate FFA follicular fluid constituent. The effects were assessed of this substitution on the Akt/GSK3β signaling pathway activity as well as the expressions of TRIB3 and FSHR at both the gene and protein levels by qPCR, Western blot and immunofluorescence staining analyses. Meanwhile, the individual effects of TRIB3 knockdown in KGN cells and p-AKT inhibitors were compared to determine the mechanisms of FFA-induced FSHR downregulation.Results: The average FSH dose consuming per oocyte (FSH dose/oocyte) was elevated and Top embryo quality ratio was decreased in women with high levels of FFAs in their follicular fluid. In these women, the GC TRIB3 and ATF4 protein expression levels were upregulated which was accompanied by FSHR downregulation. Such upregulation was confirmed based on corresponding increases in their gene expression levels. On the other hand, the levels of p-Akt decreased while p-GSK3β increased in the GCs. Moreover, TRIB3 knockdown reversed declines in FSHR expression and estradiol (E2) production in KGN cells treated with PA, which also resulted in increased p-Akt levels and declines in the p-GSK3β level. In contrast, treatment of TRIB3-knockdown cells with an inhibitor of p-Akt (Ser473) resulted in rises in the levels of both p-GSK3β as well as FSHR expression whereas E2 synthesis fell. Conclusions: During exposure to a high FFA content, TRIB3 can reduce FSHR expression through stimulation of the Akt/GSK3β pathway in human GCs. This response may contribute to inducing oocyte maturation.

Clinical efficacy and mechanism of growth hormone action in patients experiencing repeat implantation failure

2018 ◽  
Vol 96 (9) ◽  
pp. 929-932 ◽  
Author(s):  
Ye Chen ◽  
Fenghua Liu ◽  
Yingqi Nong ◽  
Jianxing Ruan ◽  
Qingqing Guo ◽  
...  
Keyword(s):  
Growth Hormone ◽  
Treatment Group ◽  
Clinical Efficacy ◽  
Granulosa Cells ◽  
Follicular Fluid ◽  
Growth Hormone Receptor ◽  
Control Group ◽  
Implantation Failure ◽  
Gh Treatment ◽  
Expression Levels

The objectives of this study were to investigate the clinical efficacy of growth hormone (GH) in patients experiencing repeat implantation failure (RIF) and explore the possible mechanism. Forty-two RIF patients undergoing in vitro fertilization – embryo transfer (IVF-ET) were enrolled in the present trial: 22 patients who received GH (treatment group) and 20 who did not receive GH (controls). The clinical pregnancy and live birth rates in the treatment group were significantly higher than those in the control group (both P < 0.05). The treatment group expressed significantly higher levels of growth hormone receptor (GHR) mRNA than the control group (P < 0.05). Further analysis showed that GH levels in follicular fluid were positively correlated with the expression levels of mRNA encoding GHR in granulosa cells (r = 0.460, P < 0.05). GH treatment enhanced IVF pregnancy outcomes and increased the expression of GHR in granulosa cells in fluid. GH levels in follicular fluid were positively correlated with the expression levels of GHR.

Effect of bovine age on the proliferative activity, global DNA methylation, relative telomere length and telomerase activity of granulosa cells

Zygote ◽  
2011 ◽  
Vol 21 (3) ◽  
pp. 256-264 ◽  
Author(s):  
Hiroya Goto ◽  
Hisataka Iwata ◽  
Shun Takeo ◽  
Keiko Nisinosono ◽  
Sayoko Murakami ◽  
...  
Keyword(s):  
Dna Methylation ◽  
Telomere Length ◽  
Granulosa Cells ◽  
Growth Phase ◽  
Genomic Dna ◽  
Proliferative Activity ◽  
Telomerase Activity ◽  
Global Dna Methylation ◽  
Relative Telomere Length ◽  
Dominant Phase

SummaryGranulosa cells influence the growth and acquisition of the developmental competence of oocytes. We investigated the effects of ageing on the proliferative activity, global genomic DNA methylation, relative telomere length and telomerase activity of bovine granulosa cells. The proliferative activity of cells was examined by bromodeoxyuridine (BrdU) assay, genomic DNA methylation was examined by enzyme-linked immunosorbent assay (ELISA), and relative telomere length and telomerase activity were examined by real-time polymerase chain reaction. We first compared the proliferative activity of the granulosa cells of the medium follicles between in dominant phase ovaries and growth phase ovaries. We observed that the proliferative activity of the granulosa cells of dominant phase ovaries was significantly lower than those of growth phase ovaries. In addition, the proliferative activity of granulosa cells was inversely associated with follicular size. Based on the results, we used granulosa cells harvested from the medium follicles (3–5 mm in diameter) on the surfaces of the dominant phase ovaries collected from cows at a slaughterhouse. The proliferative activity of the granulosa cells harvested from the ovaries of old cows (N = 8; average age 165.1 months) was lower than that of the cells from young cows (N = 8; average age 30.9 months). Global loss of cytosine methylation was detected in the granulosa cells of old cows (N = 12; average age 141.0 months) compared with young cows (N = 15; average age 27.4 months). Although the relative telomere lengths of cumulus cells were similar in the two age groups, the relative telomere lengths and telomerase activity of the granulosa cells from old cows (N = 17 and 9; average age, 164.6 and 151.3 months, respectively) tended to be shorter than those of the cells from young cows (N = 17 and 10; average age 30.6 and 28.1 months, respectively); however, this difference was not significant p = 0.09 and 0.053, respectively). In conclusion, the proliferative activity and genomic global DNA methylation significantly decreased, and the relative telomere lengths and telomerase activity of granulosa cells tended to be shorter with the age of donor cows.

scholarly journals Expression of mRNAs for pro-and anti-apoptotic factors in granulosa cells and follicular fluid of women undergoing in vitro fertilization. A pilot study

2021 ◽  
Vol 21 (1) ◽  
Author(s):  
József Bódis ◽  
Endre Sulyok ◽  
Ákos Várnagy ◽  
Viktória Prémusz ◽  
Krisztina Gödöny ◽  
...  
Keyword(s):  
Gene Expression ◽  
In Vitro Fertilization ◽  
Mrna Expression ◽  
Granulosa Cells ◽  
Follicular Fluid ◽  
Vitro Fertilization ◽  
Expression Levels ◽  
The Impact ◽  
Apoptotic Factors

Abstract Background This observational clinical study evaluated the expression levels and predictive values of some apoptosis-related genes in granulosa cells (GCs) and follicular fluid (FF) of women undergoing in vitro fertilization (IVF). Methods GCs and FF were obtained at oocyte retrieval from 31 consecutive patients with heterogeneous infertility diagnosis (age: 34.3 ± 5.8 years, body mass index: 24.02 ± 3.12 kg/m2, duration of infertility: 4.2 ± 2.1 years). mRNA expression of pro-apoptotic (BAX, CASP3, CASP8) and anti-apoptotic (BCL2, AMH, AMHR, FSHR, LHR, CYP19A1) factors was determined by quantitative RT-PCR using ROCHE LightCycler 480. Results No significant difference in GC or FF mRNA expression of pro- and anti-apoptotic factors could be demonstrated between IVF patients with (9 patients) or without (22 patients) clinical pregnancy. Each transcript investigated was detected in FF, but their levels were markedly reduced and independent of those in GCs. The number of retrieved oocytes was positively associated with GC AMHR (r = 0.393, p = 0.029), but the day of embryo transfer was negatively associated with GC LHR (r = − 0.414, p = 0.020) and GC FSHR transcripts (r = − 0.535, p = 0.002). When pregnancy positive group was analysed separately the impact of apoptosis- related gene expressions on some selected measures of IVF success could be observed. Strong positive relationship was found between gene expression levels of pro- and anti-apoptotic factors in GCs. Conclusion Our study provides only marginal evidences for the apoptosis dependence of IVF outcome and suggests that the apoptosis process induces adaptive increases of the anti-apoptotic gene expression to attenuate apoptosis and to protect cell survival.

scholarly journals TRIB3 regulates FSHR expression in human granulosa cells under high levels of free fatty acids

2021 ◽  
Vol 19 (1) ◽  
Author(s):  
Nan Wang ◽  
Chenchen Si ◽  
Lan Xia ◽  
Xian Wu ◽  
Sheng Zhao ◽  
...  
Keyword(s):  
Fatty Acids ◽  
Free Fatty Acids ◽  
Signaling Pathway ◽  
Oocyte Maturation ◽  
Granulosa Cells ◽  
Follicular Fluid ◽  
Glycogen Synthase ◽  
Obese Women ◽  
Expression Levels

Abstract Background Granulosa cells (GCs) in cumulus oophorus highly express follicle stimulating hormone receptor (FSHR), which is the most important mediator of both estradiol synthesis and oocyte maturation. Obese women have elevated free fatty acids (FFAs) levels in their follicular fluids and decreased FSHR expression in GCs, which is related to an altered protein kinase B/glycogen synthase kinase 3β (Akt/GSK3β) signaling pathway. Such FFA increases accompany 3-fold rises in pseudokinase 3 (TRIB3) expression and reduce the Akt phosphorylation status in both the human liver and in insulinoma cell lines. Therefore, in a high FFA environment, we determined if TRIB3 mediates regulation of FSHR via the Akt/GSK3β signaling pathway in human GCs. Methods GCs from women undergoing in vitro fertilization were collected and designated as high and low FFAs cohorts based on their follicular fluid FFA content. GCs with low FFA levels and a human granulosa-like tumor (KGN) cell line were exposed to palmitic acid (PA), which is a dominate FFA follicular fluid constituent. The effects were assessed of this substitution on the Akt/GSK3β signaling pathway activity as well as the expressions of TRIB3 and FSHR at both the gene and protein levels by qPCR, Western blot and immunofluorescence staining analyses. Meanwhile, the individual effects of TRIB3 knockdown in KGN cells and p-AKT inhibitors were compared to determine the mechanisms of FFA-induced FSHR downregulation. Results The average FSH dose consuming per oocyte (FSH dose/oocyte) was elevated and Top embryo quality ratio was decreased in women with high levels of FFAs in their follicular fluid. In these women, the GC TRIB3 and ATF4 protein expression levels were upregulated which was accompanied by FSHR downregulation. Such upregulation was confirmed based on corresponding increases in their gene expression levels. On the other hand, the levels of p-Akt decreased while p-GSK3β increased in the GCs. Moreover, TRIB3 knockdown reversed declines in FSHR expression and estradiol (E2) production in KGN cells treated with PA, which also resulted in increased p-Akt levels and declines in the p-GSK3β level. In contrast, treatment of TRIB3-knockdown cells with an inhibitor of p-Akt (Ser473) resulted in rises in the levels of both p-GSK3β as well as FSHR expression whereas E2 synthesis fell. Conclusions During exposure to a high FFA content, TRIB3 can reduce FSHR expression through stimulation of the Akt/GSK3β pathway in human GCs. This response may contribute to inducing oocyte maturation.

scholarly journals Relative Telomere Length and Telomerase Reverse Transcriptase (TERT) Expression Are Associated with Age in Almond (Prunus dulcis [Mill.] D.A.Webb)

Plants ◽  
2021 ◽  
Vol 10 (2) ◽  
pp. 189
Author(s):  
Katherine M. D'Amico-Willman ◽  
Elizabeth S. Anderson ◽  
Thomas M. Gradziel ◽  
Jonathan Fresnedo-Ramírez
Keyword(s):  
Reverse Transcriptase ◽  
Telomere Length ◽  
Telomerase Reverse Transcriptase ◽  
Perennial Crop ◽  
Related Disorder ◽  
Age Cohorts ◽  
Biomarkers Of Aging ◽  
Relative Telomere Length ◽  
Age Related ◽  
Tert Expression

While all organisms age, our understanding of how aging occurs varies among species. The aging process in perennial plants is not well-defined, yet can have implications on production and yield of valuable fruit and nut crops. Almond exhibits an age-related disorder known as non-infectious bud failure (BF) that affects vegetative bud development, indirectly affecting kernel yield. This species and disorder present an opportunity to address aging in a commercially relevant and vegetatively propagated perennial crop. The hypothesis tested in this study was that relative telomere length and/or telomerase reverse transcriptase (TERT) expression can serve as biomarkers of aging in almond. Relative telomere lengths and expression of TERT, a subunit of the enzyme telomerase, were measured via qPCR methods using bud and leaf samples collected from distinct age cohorts over a two-year period. Results from this work show a marginal but significant association between both relative telomere length and TERT expression, and age, suggesting that as almonds age, telomeres shorten and TERT expression decreases. This work provides information on potential biomarkers of perennial plant aging, contributing to our knowledge of this process. In addition, these results provide opportunities to address BF in almond breeding and nursery propagation.

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