Published 2020-01-22
Fadil Pramudhya Hoesain Doddy M. Soebadi Wahjoe Djatisoesanto


Objective: To analyze the differences of the number of Leydig, spermatogonium, and Sertoli cells in mice, after being given nicotine inhalation, and the effect of recovery when nicotine stopped. Materials & Methods: This is experimental studies with a post-test only control group design. The sample used was 36 adult male mice (10-12 weeks) (Rattus Norvegicus) Sprague-Dawley strain, which were divided into three groups. P1 group was given nicotine inhalation at a dose of 4 mg/kg/day for 15 days, P2 group was given nicotine inhalation at a dose of 4 mg/kg/day for 15 days and was free of treatment for 15 days. The control group (NC) has no treatment at all. Histological examination and calculation of Sertoli cells, Leydig cell, and spermatogonium were processed within 1 hour after terticular sample collection. Then carried out to statistical analysis. 100x and 400x magnification is used to obtain the histopathological. Results: Post-hoc LSD test for each variable (Leydig cells, Sertoli cells, and spermatogonium), showed that the NC group had a significantly higher number compared to group P1. P2 group has a significantly higher number compared to group P1. There is no significant difference between the NC group and the P2 group. There was a structural difference in the sample testicles which exposed to nicotine. Conclusion: Nicotine exposure with a dose of 4 mg/kg/day for 15 days has a significant effect on decreasing the number of Leydig cells, spermatogonium cells, and Sertoli cells in rats and giving a free-treatment period for 15 days, giving the testis time to do recovery and regeneration (the reversible damage of testicular structure).



nicotine, Leydig cells, Sertoli cells, spermatogonium, reversible damage


WHO. WHO Manual for the Standardized Investigation and Diagnosis of the Infertile Couple. Cambridge: Cambridge University Press; 2000.

Dohle GR, Colpi GM, Hargreave TB, Papp GK, Jungwirth A, Weidner W, et al. EAU guidelines on male infertility. Eur Urol. 2018; 48(5): 703–11.

Purnomo BB. Infertilitas pada Pria. In: Dasar-Dasar Urologi. 3rd ed. Jakarta; 2011. p. 305–17.

Pusat Data dan Informasi Kementrian Kesehatan Indonesia. Perilaku Merokok Masyarakat Indonesia; 2015.

Harlev A. Smoking and Male Infertility : An Evidance-Based Review. Smok Male Infertil An Evidance-Based Rev. 2015; 33.

Mahanem MN, Nor-Asmaniza AB, Phang HT, Muhammad HR. Effects of Nicotine and Co-Administration of Nicotine and Vitamin E on Testis and Sperm Quality of Adult Rats. 2006; 35(2): 47–52.

Ahmadnia H, Ghanbari M, Moradi MR, Khaje-Dalouee M. Effect of cigarette smoke on spermatogenesis in rats. Urol J. 2007; 4(3): 159–63.

Oyeyipo IP, Maartens PJ, du Plessis SS. In vitro effects of nicotine on human spermatozoa. Andrologia. 2014; 46(8): 887-92.

Nesseim WH, Haroun HS, Mostafa E, Youakim MF, Mostafa T. Effect of nicotine on spermatogenesis in adult albino rats. Andrologia. 2011; 43(6): 398–404.

Rizaldi A, Soebadi DM, Soetojo. Pengaruh Paparan Kronis Nikotin secara Inhalasi terhadap Jumlah Sel Spermatogonium, Sel Sertoli dan Sel Leydig Tikus Putih Strain Wistar Usia Muda (Penelitian Laboratoris Eksperimental). Univ Airlangga. 2018.

Balai Pengujian Hewan. Teknis Pemeliharaan Hewan Percobaan Untuk Pengujian Mutu Obat Hewan. Jakarta: Departemen Pertanian; 1989. p. 8–11.

Dahlan S. Statistik untuk kedokteran dan kesehatan: Uji hipotesis. Jakarta: Bina Mitra Press; 2006.

Snedecor G. Ten thousand randomly assorted digits. In: Statistical Methods. Iowa: The Iowa State University Press; 1974. p. 543.

Henkel RR. Leukocytes and oxidative stress: dilemma for sperm function and male fertility. Asian J Androl. 2011 Jan; 13(1): 43–52.

Aydos K, Güven MC, Can B, Ergün A. Nicotine toxicity to the ultrastructure of the testis in rats. BJU Int. 2001; 88(6): 622–6.

Jana K, Samanta PK, Kumar De D. Nicotine diminishes testicular gametogenesis, steroidogenesis, and steroidogenic acute regulatory protein expression in adult albino rats: Possible influence on pituitary gonadotropins and alteration of testicular antioxidant status. Toxicol Sci. 2010; 116(2): 647–59.

Agarwal A, Saleh RA, Bedaiwy MA. Role of reactive oxygen species in the pathophysiology of human reproduction. Fertil Steril. 2003 Apr; 79(4): 829–43.

Esteves SC. Effect of cigarette smoking on levels of seminal oxidative stress in infertile men: a prospective study. Int Braz J Urol. 2002; 28(5): 484–5.

Kanwal S, Ameer MK, Mehboob F. Toxic effect of nicotine on leydig cell count & testosterone levels in adult albino mice & its protection by date palm pit powder. Pakistan J Med Heal Sci. 2016; 10(4): 1356–9.

Kim KH, Joo KJ, Park HJ, Kwon CH, Jang MH, Kim CJ. Nicotine induces apoptosis in TM3 mouse Leydig cells. Fertil Steril. 2005; 83(4 SUPPL.): 1093–9.

Mosadegh M, Hasanzadeh S, Razi M. Nicotine-induced damages in testicular tissue of rats; evidences for bcl-2, p53 and caspase-3 expression. Iran J Basic Med Sci. 2017; 20(2): 199–208.

Asl SS, Farhadi MH, Moosavizadeh K, Saraei ASK, Soleimani M, Jamei SB, et al. Evaluation of Bcl-2 family gene expression in hippocampus of 3, 4-methylenedioxymethamphetamine treated rats. Cell J. 2012; 13(4): 275–80.

Susan E. Apoptosis: A Reveiw of Programmed Cell Death. Toxicol Pathol. 2007; 35(4): 496–516.

Saleh RA, Agarwal A, Sharma RK, Nelson DR, Thomas AJ. Effect of cigarette smoking on levels of seminal oxidative stress in infertile men: a prospective study. Fertil Steril. 2002; 78(3): 491–9.

Budin SB, Kho JH, Lee JH, Ramalingam A, Jubaidi FF, Latif ES, et al. Low-dose Nicotine Exposure Induced the Oxidative Damage of Reproductive Organs and Altered the Sperm Characteristics of Adolescent Male Rats. Malaysian J Med Sci. 2017; 24(6): 50–7.

Oyeyipo IP, Raji Y, Emikpe BO, Bolarinwa AF. Effects of nicotine on sperm characteristics and fertility profile in adult male rats: A possible role of cessation. J Reprod Infertil. 2011; 12(3): 201–7.

Oyeyipo IP, Raji Y, Bolarinwa AF. Nicotine alters male reproductive hormones in male albino rats: The role of cessation. J Hum Reprod Sci. 2013; 6(1): 40–4.

Copyright Information
Department of Urology, Faculty of Medicine/Airlangga University