Melatonin levels and microRNA (miRNA) relative expression profile in the follicular ambient microenvironment in patients
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REPRODUCTIVE PHYSIOLOGY AND DISEASE
Melatonin levels and microRNA (miRNA) relative expression profile in the follicular ambient microenvironment in patients undergoing in vitro fertilization process Haroon Latif Khan 1 & Shahzad Bhatti 1,2,3,4 & Sana Abbas 1 & Celal Kaloglu 5 & Syeda Qurat-ul-Ain Zahra 6 & Yousaf Latif Khan 1,7 & Zahira Hassan 8 & Nilgün Öztürk Turhan 9 & Hikmet Hakan Aydin 10 Received: 23 July 2020 / Accepted: 9 November 2020 # Springer Science+Business Media, LLC, part of Springer Nature 2020
Abstract Purpose Intrafollicular fluid (IFF) melatonin plays a decisive role in maintaining granulosa cells’ DNA integrity and protects them against apoptosis. It reduces oxidative stress and improves the oocyte quality with a higher fertilization rate. Method This prospective study investigated the antioxidant property of IFF melatonin and its impact on IVF outcome parameters. We also explored the relative expression of five microRNAs (miR-663b, miR-320a, miR-766-3p, miR-132-3p, miR-16-5p) and levels of cell-free DNA (cfDNA) by real-time PCR in unexplained infertile patients. We collected 425 follicular fluid (FF) samples containing mature oocytes from 295 patients undergoing IVF. Results Patients were subgrouped based on IFF melatonin concentration (group A ≤ 30 pg/mL, group B > 70 to ≤ 110 pg/mL, group C > 111 to ≤ 385 pg/mL). Our results showed that patients with ≤ 30 pg/mL IFF melatonin levels have significantly higher oxidative stress markers, cfDNA levels, and lower relative expression of miR-663b, miR-320a, miR-766-3p, miR-132-3p, and miR-16-5p compared to other subgroups (p < 0.001). Similarly, they have a low fertilization rate and a reduced number of highquality day 3 embryos. Conclusion Findings suggest that the therapeutic use of melatonin produces a considerable rise in the number of mature oocytes retrieved, fertilization rate, and good-quality embryo selection. Furthermore, miRNA signature enhances the quality of embryo selection, thus, may allow us to classify them as non-invasive biomarkers to identify good-quality embryos. Keywords Melatonin . miRNA expression . Cell-free DNA . Follicular fluid . Embryo quality . Unexplained infertility
Introduction Infertility is a non-contracepting couple’s inability to achieve a clinical pregnancy after a year or more of regular unprotected sexual intercourse [1]. Infertility is said to be idiopathic when * Shahzad Bhatti [email protected]; [email protected] 1
Lahore Institute of Fertility and Endocrinology, Hameed Latif Hospital, 14-Abu Bakar Block, New Garden Town, Lahore 54800, Pakistan
2
Department of Human Genetics and Molecular biology, University of Health Sciences, Lahore 54600, Pakistan
3
Department of Medical Education, Rashid Latif Medical College, Lahore, Pakistan
4
Department of Human Genetics and Molecular Biology, University of Health Sciences, Lahore 54800, Pakistan
standard investigation such as tests for ovulation, tubal patency, and semen quality has not been seen abnormal, and no pathological condition is
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