The therapeutic effect of melatonin on female offspring ovarian reserve and quality in BALB/C mice after exposing their mother to methamphetamine during pregnancy and lactation

Document Type : Original Article


1 Department of Anatomy and Cell Biology, School of Medicine, Mashhad University of Medical Sciences, Mashhad, Iran

2 Medical Genetics and Molecular Medicine Department, School of Medicine, Mashhad University of Medical Sciences, Mashhad, Iran

3 Medical Genetics Research Center, Mashhad University of Medical Sciences, Mashhad, Iran

4 Department of Medical Biotechnology and Nanotechnology, Faculty of Medicine, Mashhad University of Medical Sciences, Mashhad, Iran

5 Applied Biomedical Research Center, School of Medicine, Mashhad University of Medical Sciences, Mashhad, Iran


Objective(s): Nowadays, methamphetamine (METH) abuse as a psychotropic drug is increasing. There is insufficient information about its adverse effects on the ovarian reserve of the next generation. Herein, we tried to investigate the effect of METH abuse during pregnancy and lactation and, subsequently, the therapeutic effect of melatonin on ovarian reserve in offspring.
Materials and Methods: In the present study, BALB/C pregnant female mice were divided into 3 groups: Control, Saline, and METH (5mg/Kg). METH was injected during pregnancy and lactation, and the female offspring of each group was divided into 2 subgroups: A) treated with 10 mg/kg Melatonin daily until puberty (6 weeks old) and B) received distilled water. The animals were sacrificed at 6 weeks of age, and blood samples were collected for hormonal assessments; the right ovaries were removed and fixed for TUNEL and Hematoxylin & Eosin staining, and the left ovaries were removed and stored for gene expression and oxidative stress evaluation.
Results: In the MTEH group, two indicators of ovarian reserve (including anti-Müllerian hormone (AMH) and primordial follicle, and Cyclin D1 (CCND-1) and Proliferating Cell Nuclear Antigen (PCNA) genes expression significantly decreased, and the oxidative stress and apoptosis significantly increased in comparison with other groups. After lactation in the MTEH group, melatonin treatment significantly improved the ovarian reserve and gene expression and declined apoptosis and oxidative stress.
Conclusion: METH abuse during pregnancy and lactation decreased ovarian reserve in offspring. The administration of melatonin as an anti-oxidant agent after lactation can counteract the adverse effects of METH on offspring ovaries.


1. Limanaqi F, Gambardella S, Biagioni F, Busceti CL, Fornai F. Epigenetic effects induced by methamphetamine and methamphetamine-dependent oxidative stress. Oxid Med Cell Longev 2018; 2018: 4982453.
2. Wang L, Qu G, Dong X, Huang K, Kumar M, Ji L, et al. Long-term effects of methamphetamine exposure in adolescent mice on the future ovarian reserve in adulthood. Toxicol Lett 2016; 242: 1-8.
3. Prakash MD, Tangalakis K, Antonipillai J, Stojanovska L, Nurgali K, Apostolopoulos V. Methamphetamine: effects on the brain, gut and immune system. Pharmacol Res 2017; 120: 60-67.
4. Nudmamud-Thanoi S, Sueudom W, Tangsrisakda N, Thanoi S. Changes of sperm quality and hormone receptors in the rat testis after exposure to methamphetamine. Drug Chem Toxicol 2016; 39: 432-438.
5. Gorman MC, Orme KS, Nguyen NT, Kent EJ 3rd, Caughey AB. Outcomes in pregnancies complicated by methamphetamine use. Am J Obstet Gynecol 2014; 211: 429 e421-427.
6. Shen WW, Zhang YS, Li LH, Liu Y, Huang XN, Chen LH, et al. Long-term use of methamphetamine disrupts the menstrual cycles and hypothalamic-pituitary-ovarian axis. J Addict Med 2014; 8: 183-188.
7. Baei F, Rajabzadeh A, Bagheri J, Jalayeri Z, Ebrahimzadeh-Bideskan A. Effect of methamphetamine exposure during pregnancy and lactation on polysialic acid-neural cell adhesion molecule expression in rat’s offspring hippocampus. Metab Brain Dis 2017; 32: 991-1002.
8. Nezhad SM, Ghaffari NM, Fadaee FF, Salehi M, Salimi M, Shams MZ, et al. The effect of methamphetamine on oocyte quality, fertilization rate and embryo development in mice.  2016; 4: 8-12.
9. Agarwal A, Aponte-Mellado A, Premkumar BJ, Shaman A, Gupta S. The effects of oxidative stress on female reproduction: a review. Reprod Biol Endocrinol 2012; 10: 1-31.
10. Potula R, Hawkins BJ, Cenna JM, Fan S, Dykstra H, Ramirez SH, et al. Methamphetamine causes mitrochondrial oxidative damage in human T lymphocytes leading to functional impairment. J Immunol 2010; 185: 2867-2876.
11. Lv PP, Jin M, Rao JP, Chen J, Wang LQ, Huang CC, et al. Role of anti-Mullerian hormone and testosterone in follicular growth: A cross-sectional study. BMC Endocr Disord 2020; 20: 1-10.
12. Wang S, He G, Chen M, Zuo T, Xu W, Liu X. The role of anti-oxidant enzymes in the ovaries. Oxid Med Cell Longev 2017; 2017: 4371714.
13. Wongprayoon P, Govitrapong P. Melatonin attenuates methamphetamine-induced neurotoxicity. Curr Pharm Des 2016; 22: 1022-1032.
14. Reiter RJ, Mayo JC, Tan DX, Sainz RM, Alatorre‐Jimenez M, Qin L. Melatonin as an anti-oxidant: Under promises but over delivers. J Pineal Res 2016; 61: 253-278.
15. Song C, Peng W, Yin S, Zhao J, Fu B, Zhang J, et al. Melatonin improves age-induced fertility decline and attenuates ovarian mitochondrial oxidative stress in mice. Sci Rep 2016; 6: 1-15.
16. Saberi A, Sepehri G, Safi Z, Razavi B, Jahandari F, Divsalar K, et al. Effects of methamphetamine on testes histopathology and spermatogenesis indices of adult male rats. Addict Health 2017; 9: 199-205.
17. Ahmadi M, Rostamzadeh A, Fathi F, Mohammadi M, Rezaie MJ. The effect of melatonin on histological changes of ovary in induced polycystic ovary syndrome model in mice. Middle East Fertil Soc J 2017; 22: 255-259.
18. Zhang C, Wu J, He J, Liu X, Chen X, Tong C, et al. The regulation of high insulin levels on ovary apoptosis in early pregnant mice. Biochem Biophys Res Commun 2017; 483: 786-792.
19. Ghorbani A, Feizpour A, Hashemzahi M, Gholami L, Hosseini M, Soukhtanloo M, et al. The effect of adipose derived stromal cells on oxidative stress level, lung emphysema and white blood cells of guinea pigs model of chronic obstructive pulmonary disease. DARU Journal of Pharmaceutical Sciences 2014; 22: 22-26.
20. Jalilvand N, Hosseini M, Beheshti F, Ebrahimzadeh-Bideskan A. Protective effect of PPARγ agonist pioglitazone, on testicular tissue and sperm parameters in hypothyroid rats. Toxin Reviews 2021; 40: 267-276.
21. Gaharwar US, Meena R, Rajamani P. Iron oxide nanoparticles induced cytotoxicity, oxidative stress and DNA damage in lymphocytes. J Appl Toxicol 2017; 37: 1232-1244.
22.Karimi S, Tabatabaei SN, Gutleb AC, Ghaffari Novin M, Ebrahimzadeh-Bideskan A, Shams Mofarahe Z. The effect of PEGylated iron oxide nanoparticles on sheep ovarian tissue: An ex-vivo nanosafety study. Heliyon 2020; 6: e04862.
23. Ross MH, Pawlina W. Histology: A text and atlas: with Correlated Cell and Molecular Biology: Wolters Kluwer Health; 2016.
24. Mobaraki F, Seghatoleslam M, Fazel A, Ebrahimzadeh-Bideskan A. Effects of MDMA (ecstasy) on apoptosis and heat shock protein (HSP70) expression in adult rat testis. Toxicol Mech Methods 2018; 28: 219-229.
25. Rajabzadeh AA, Bideskan AR, Haghir H, Fazel AR. Morphometrical study of polysialylated neural cell adhesion molecule positive cells in rat pups hippocampus following induction of seizure during pregnancy. Iran Biomed J 2011; 15: 157-163.
26. Pashaei-Asl F, Pashaei-Asl R, Khodadadi K, Akbarzadeh A, Ebrahimie E, Pashaiasl M. Enhancement of anticancer activity by silibinin and paclitaxel combination on the ovarian cancer. Artif Cells Nanomed Biotechnol 2018; 46: 1483-1487.
27. Li S, Gu H, Huang Y, Peng Q, Zhou R, Yi P, et al. Circular RNA 101368/miR-200a axis modulates the migration of hepatocellular carcinoma through HMGB1/RAGE signaling. Cell Cycle 2018; 17: 2349-2359.
28. Rudzinskas SA, Williams KM, Mong JA, Holder MK. Sex, drugs, and the medial amygdala: a model of enhanced sexual motivation in the female rat. Front Behav Neurosci 2019; 13: 1-13.
29.Dinger J, Hinner P, Reichert J, Rudiger M. Methamphetamine consumption during pregnancy - effects on child health. Pharmacopsychiatry 2017; 50: 107-113.
30. Kandemir YB, Aydin C, Gorgisen G. The effects of melatonin on oxidative stress and prevention of primordial follicle loss via activation of mTOR pathway in the rat ovary. Cell Mol Biol 2017; 63: 100-106.
31. Findlay JK, Hutt KJ, Hickey M, Anderson RA. How is the number of primordial follicles in the ovarian reserve established? Biol Reprod 2015; 93:111, 111-117.
32. de Angelis C, Nardone A, Garifalos F, Pivonello C, Sansone A, Conforti A, et al. Smoke, alcohol and drug addiction and female fertility. Reprod Biol Endocrinol 2020; 18: 1-26.
33. Tamura H, Kawamoto M, Sato S, Tamura I, Maekawa R, Taketani T, et al. Long-term melatonin treatment delays ovarian aging. J Pineal Res 2017; 62:e 12381.
34. Pankhurst MW. A putative role for anti-Mullerian hormone (AMH) in optimising ovarian reserve expenditure. J Endocrinol 2017; 233:R1-R13.
35. Ma M, Chen XY, Li B, Li XT. Melatonin protects premature ovarian insufficiency induced by tripterygium glycosides: Role of SIRT1. Am J Transl Res 2017; 9: 1580-1602.
36. Xiong Q, Ru Q, Tian X, Zhou M, Chen L, Li Y, et al. Krill oil protects PC12 cells against methamphetamine-induced neurotoxicity by inhibiting apoptotic response and oxidative stress. Nutr Res 2018; 58: 84-94.
37.Shin EJ, Tran HQ, Nguyen PT, Jeong JH, Nah SY, Jang CG, et al. Role of mitochondria in methamphetamine-induced dopaminergic neurotoxicity: involvement in oxidative stress, neuroinflammation, and pro-apoptosis-a review. Neurochem Res 2018; 43: 66-78.
38. Li Y, Hu Z, Chen B, Bu Q, Lu W, Deng Y, et al. Taurine attenuates methamphetamine-induced autophagy and apoptosis in PC12 cells through mTOR signaling pathway. Toxicol Lett 2012; 215: 1-7.
39. Redza-Dutordoir M, Averill-Bates DA. Activation of apoptosis signalling pathways by reactive oxygen species. Biochim Biophys Acta 2016; 1863: 2977-2992.
40. Yang Q, Zhu J, Luo X, Li F, Cong L, Wang Y, et al. Melatonin attenuates cadmium-induced ovulatory dysfunction by suppressing endoplasmic reticulum stress and cell apoptosis. Reprod Biol Endocrinol 2019; 17: 1-8.
41. Delkhosh A, Delashoub M, Tehrani AA, Bahrami AM, Niazi V, Shoorei H, et al. Upregulation of FSHR and PCNA by administration of coenzyme Q10 on cyclophosphamide-induced premature ovarian failure in a mouse model. J Biochem Mol Toxicol 2019; 33: e22398.
42. Tyasi TL, Sun X, Shan X, Liswaniso S, Chimbaka IM, Qin N, et al. Effects of RAC1 on proliferation of hen ovarian prehierarchical follicle granulosa cells. Animals (Basel) 2020; 10:1-17.