Oxidative stress and metformin: An in-vitro study on serum and primary human granulosa cell cultures
More details
Hide details
PAPRSB Institute of Health Sciences, Universiti Brunei Darussalam, BRUNEI
Department of Biological and Biomedical Sciences, Aga Khan University, Karachi, PAKISTAN
Australian Concept Infertility Medical Centre, Karachi, PAKISTAN
Publication date: 2022-04-20
Electron J Gen Med 2022;19(4):em381
To observe the effect of metformin on the oxidative stress (OS) levels and SIRT1 expression in primary human granulosa cell cultures from infertile females.

This cross-sectional study was conducted from August 2017-July 2019 on ten infertile patients. After written informed consent, the follicular fluid of these subjects was collected for establishing primary human granulosa cell cultures (HGCs), to test the capacity of metformin to reduce OS. The cultured HGCs were divided into (i) control: HGCs without any treatment, (ii) test1: HGCs induced with H2O2 (metformin absent), and (iii) test2: HGCs treated with metformin after H2O2 induction. Baseline OS (control group) was estimated, it was induced and measured by an increase in optical density (OD) (test1 & test 2). Finally, test2 was incubated with metformin (1 ml of 100 mmol/l concentration) and it effect on the levels of OS were determined by Mishra OS assay. Effect of metformin modulating SIRT1 expression in OS-induced cells was analyzed using quantitative-PCR.

Age of female subjects was 32.04±2.29years and BMI was 27.61±2.15 kg/m2. Test (OS induced) samples gave an OD of 0.28 (0.16-0.40) while control HGC samples gave an OD; 0.153 (0.09-0.23). Test cells showed significant reduction in ODs after metformin treatment. The relative expression of SIRT1 in metformin untreated and treated cells was found to be 61.5% and 80%, respectively.

Metformin was found to suppress OS in HGCs and increases the expression of SIRT1 in OS induced environment of primary cell cultures, suggesting a relationship between metformin, SIRT1 expression, and reduction of OS.

Alam F, Khan T A, Ali R, Tariq F, Rehman R. SIRTI and cortisol in unexplained infertile females; a cross sectional study, in Karachi Pakistan. Taiwan J Obstet Gynecol. 2020;59(2):189-94. https://doi.org/10.1016/j.tjog... PMid:32127136.
Geraghty P. Each woman’s menopause: An evidence based resource. Springer; 2022. https://doi.org/10.1007/978-3-....
Holmes DT, Kline G. The endocrinology of aging. In: Winter WE, Holmquist B, Sokoll LJ, Bertholf RL, editors. Handbook of diagnostic endocrinology. Elsevier; 2021. p. 663-85. https://doi.org/10.1016/B978-0....
Gualtieri R, Kalthur G, Barbato V, Di Nardo M, Adiga SK, Talevi R. Mitochondrial dysfunction and oxidative stress caused by cryopreservation in reproductive cells. Antioxidants (Basel). 2021;10(3):337. https://doi.org/10.3390/antiox... PMid:33668300 PMCid:PMC7996228.
Hardy MLM, Day ML, Morris MB. Redox regulation and oxidative stress in mammalian oocytes and embryos developed in vivo and in vitro. Int J Environ Res Public Health. 2021;18(21):11374. https://doi.org/10.3390/ijerph... PMid:34769890 PMCid:PMC8583213.
Nath B, Roy H. Antioxidants in female reproductive biology. In: Waisundara VY, editor. Antioxidants-Benefits, sources, mechanisms of action. 2021. https://doi.org/10.5772/intech....
Tatone C, Di Emidio G, Barbonetti A, et al. Sirtuins in gamete biology and reproductive physiology: emerging roles and therapeutic potential in female and male infertility. Hum Reprod Update. 2018;24(3):267-89. https://doi.org/10.1093/humupd... PMid:29447380.
LaMoia TE, Shulman GI. Cellular and molecular mechanisms of metformin action. Endocr Rev. 2021;42(1):77-96. https://doi.org/10.1210/endrev... PMid:32897388 PMCid:PMC7846086.
Hunter RW, Hughey CC, Lantier L, et al. Metformin reduces liver glucose production by inhibition of fructose-1-6-bisphosphatase. Nat Med. 2018;24(9):1395-406. https://doi.org/10.1038/s41591... PMid:30150719 PMCid:PMC6207338.
Flory J, Lipska K. Metformin in 2019. JAMA. 2019;321(19):1926-7. https://doi.org/10.1001/jama.2... PMid:31009043 PMCid:PMC7552083.
Podhorecka M. Metformin-its anti-cancer effects in hematologic malignancies. Oncol Rev. 2021;15(1):514. https://doi.org/10.4081/oncol.... PMid:33747367 PMCid:PMC7967492.
Bhatti GK, Gupta A, Reddy PH, Bhatti JS. Translation of bioenergetics therapies: Challenges from bench to bedside. Clin Bioenerg. 2021;489-507. https://doi.org/10.1016/b978-0....
Rehman R, Abidi SH, Alam F. Metformin, oxidative stress, and infertility: A way forward. Front Physiol. 2018;9:1722. https://doi.org/10.3389/fphys.... PMid:30555352 PMCid:PMC6281994.
Calabrese EJ, Kozumbo WJ. The hormetic dose-response mechanism: Nrf2 activation. Pharmacol Res. 2021;167:105526. https://doi.org/10.1016/j.phrs... PMid:33667690.
Xie Y, Wang L, Khan M A, et al. Metformin and androgen receptor-axis-targeted (ARAT) agents induce two PARP-1-dependent cell death pathways in androgen-sensitive human prostate cancer cells. Cancers (Basel). 2021;13(4):633. https://doi.org/10.3390/cancer... PMid:33562646 PMCid:PMC7914929.
Deng M, Su D, Xu S, et al. Metformin and vascular diseases: A focused review on smooth muscle cell function. Front Pharmacol. 2020;11:635. https://doi.org/10.3389/fphar.... PMid:32457625 PMCid:PMC7227439.
Horakova O, Kroupova P, Bardova K, et al. Metformin acutely lowers blood glucose levels by inhibition of intestinal glucose transport. Sci Rep. 2019;9(1):6156. https://doi.org/10.1038/s41598... PMid:30992489 PMCid:PMC6468119.
Uckan K, Demir H, Turan K, Sarikaya E, Demir C. Role of oxidative stress in obese and nonobese PCOS patients. Int J Clin Pract. 2022;2022:4579831. https://doi.org/10.1155/2022/4....
Palomba S, Falbo A, La Sala GB. Effects of metformin in women with polycystic ovary syndrome treated with gonadotrophins for in vitro fertilisation and intracytoplasmic sperm injection cycles: A systematic review and meta-analysis of randomised controlled trials. BJOG. 2013;120(3):267-76. https://doi.org/10.1111/1471-0... PMid:23194199.
Costello MF, Chapman M, Conway U. A systematic review and meta-analysis of randomized controlled trials on metformin co-administration during gonadotrophin ovulation induction or IVF in women with polycystic ovary syndrome. Hum Reprod. 2006;21(6):1387-99. https://doi.org/doi:10.1093/hu... PMid:16449310.
Ashraf M, Mustansir F, Baqir SM, Alam F, Rehman R. Changes in vitamin E levels as a marker of female infertility. J Pak Med Assoc. 2020;70(10):1762-6. https://doi.org/10.5455/JPMA.4... PMid: 3159749.
Upadhyay S, Vaish S, Dhiman M. Hydrogen peroxide-induced oxidative stress and its impact on innate immune responses in lung carcinoma A549 cells. Mol Cell Biochem. 2019;450(1-2):135-47. https://doi.org/10.1007/s11010... PMid:29938378.
Campos-Shimada LB, Hideo Gilglioni E, Fernandes Garcia R, Rizato Martins-Maciel E, Luiza Ishii-Iwamoto E, Luzia Salgueiro-Pagadigorria C. Superoxide dismutase: A review and a modified protocol for activities measurements in rat livers. Arch Physiol Biochem. 2020;126(4):292-9. https://doi.org/10.1080/138134... PMid:30372625.
Shentu X-C, Ping X-Y, Cheng Y-L, Zhang X, Tang Y-L, Tang X-J. Hydrogen peroxide-induced apoptosis of human lens epithelial cells is inhibited by parthenolide. Int J Ophthalmol. 2018;11(1):12-7. https://doi.org/10.18240/ijo.2... PMid:29375984 PMCid:PMC5767651.
Li X-H, Li D, Liu C, Zhang M-M, Guan X-J, Fu Y-P. p33ING1b regulates acetylation of p53 in oral squamous cell carcinoma via SIR2. Cancer Cell Int. 2020;20:398. https://doi.org/10.1186/s12935... PMid:32831651 PMCid:PMC7436958.
Alam F, Khan TA, Amjad S, Rehman R. Association of oxidative stress with female infertility-A case control study. J Pak Med Assoc. 2019;69(5):627-31. PMid:31105280.
Jamilian H, Jamilian M, Foroozanfard F, Afshar Ebrahimi F, Bahmani F, Asemi Z. Comparison of myo-inositol and metformin on mental health parameters and biomarkers of oxidative stress in women with polycystic ovary syndrome: A randomized, double-blind, placebo-controlled trial. J Psychosom Obstet Gynecol. 2018;39(4):307-14. https://doi.org/10.1080/016748... PMid:28980870.
Walters KA, Bertoldo MJ, Handelsman DJ. Evidence from animal models on the pathogenesis of PCOS. Best Pract Res Clin Endocrinol Metab. 2018;32(3):271-81. https://doi.org/10.1016/j.beem... PMid:29779581.
Raperport C, Chronopoulou E, Homburg R. Effects of metformin treatment on pregnancy outcomes in patients with polycystic ovary syndrome. Expert Rev Endocrinol Metab. 2021;16(2):37-47. https://doi.org/10.1080/174466... PMid:33634727.
Pericuesta E, Laguna-Barraza R, Ramos-Ibeas P, et al. D-chiro-inositol treatment affects oocyte and embryo quality and improves glucose intolerance in both aged mice and mouse models of polycystic ovarian syndrome. Int J Mol Sci. 2020;21(17):6049. https://doi.org/10.3390/ijms21... PMid:32842637 PMCid:PMC7504697.
Bost F, Rena G, Viollet B. Metformin: Beyond diabetes. Front Endocrinol (Lausanne). 2019;10:851. https://doi.org/10.3389/fendo.... PMid:31866949 PMCid:PMC6909380.
Cuyàs E, Verdura S, Llorach-Parés L, et al. Metformin is a direct SIRT1-activating compound: Computational modeling and experimental validation. Front Endocrinol (Lausanne). 2018;9:657. https://doi.org/10.3389/fendo.... PMid:30459716 PMCid:PMC6232372.
Kosgei VJ, Coelho D, Guéant-Rodriguez R-M, Guéant J-L. Sirt1-PPARS cross-talk in complex metabolic diseases and inherited disorders of the one carbon metabolism. Cells. 2020;9(8):1882. https://doi.org/10.3390/cells9... PMid:32796716 PMCid:PMC7465293.
Torres W, Nava M, Galbán N, et al. Anti-aging effect of metformin: A molecular and Therapeutical perspective. Curr Pharm Des. 2020;26(35):4496-508. https://doi.org/10.2174/138161... PMid:32674728.
Kamalipour F, Jalali H, Azarnia M. Comparison the effect of metformin and clomiphene citrate on sirtuin3 gene expression in the oocytes of mice with polycystic ovary syndrome. Iran J Pharm Res. 2020;19(4):160-8. https://doi.org/10.22037/ijpr.... PMid:33841532 PMCid:PMC8019885.
Huang C-C, Chou C-H, Yang Y-S, et al. Metformin: A novel promising option for fertility preservation during cyclophosphamide-based chemotherapy. Mol Hum Reprod. 2021;27(1):gaaa084. https://doi.org/10.1093/molehr... PMid:33543290 PMCid:PMC8494485.
Journals System - logo
Scroll to top