Nuclear Factor Kappa-B Protein Levels in Sperm of Obese Men with and without Diabetes; Cellular Approach in Male Infertility

Document Type : Original Article


1 Reproductive Epidemiology Research Center, Royan Institute for Reproductive Biomedicine, ACECR, Tehran, Iran

2 Department of Embryology, Reproductive Biomedicine Research Center, Royan Institute for Reproductive Biomedicine, ACECR, Tehran, Iran

3 Department of Andrology, Reproductive Biomedicine Research Center, Royan Institute for Reproductive Biomedicine, ACECR, Tehran, Iran

4 Department of Basic Science, Faculty of Veterinary Medicine, University of Tehran, Tehran, Iran

5 Department of Theriogenology, Faculty of Veterinary Medicine, University of Tehran, Tehran, Iran


Although the role of obesity and diabetes mellitus (DM) in male infertility is well established, little information
about the underlying cellular mechanisms in infertility is available. In this sense, nuclear factor kappa-B (NF-kB) has
been recognized as an important regulator in obesity and DM; However, its function in the pathogenesis of male
infertility has never been studied in obese or men who suffer from diabetes. Therefore, the main goal of current research
is assessing NF-kB existence and activity in ejaculated human spermatozoa considering the obesity and diabetics
condition of males.

Materials and Methods:
In an experimental study, the ELISA technique was applied to analyze NF-kB levels in sperm
of four experimental groups: non-obese none-diabetic men (body mass index (BMI) <25 kg/m2; control group; n=30),
obese non-diabetic men (BMI >30 kg/m2; OB group; n=30), non-obese diabetic men (BMI <25 kg/m2; DM group; n=30),
and obese diabetic men (BMI >30 kg/m2; OB-DM group; n=30) who were presented to Royan Institute Infertility Center.
In addition, protein localization was shown by Immunocytofluorescent assay. Sperm features were also evaluated using

The diabetic men were older than non-diabetic men regardless of obesity status (P=0.0002). Sperm progressive
motility was affected by obesity (P=0.035) and type A sperm progressive motility was affected by DM (P=0.034). The
concentration of sperm (P=0.013), motility (P=0.025) and morphology (P<0.0001) were altered by obesity × diabetes
interaction effects. The NF-kB activity was negatively influenced by the main impact of diabetics (P=0.019). Obesity
did not affect (P=0.248) NF-kB activity. Uniquely, NF-kB localized to the midpiece of sperm and post-acrosomal areas.

The current study indicated a lower concentration of NF-kB in diabetic men, no effect of obesity on NF-kB
was observed yet. Additionally, we revealed the main obesity and diabetes effects, and their interaction effect adversely
influenced sperm characteristics.


1. Zhong O, Ji L, Wang J, Lei X, Huang H. Association of diabetes and obesity with sperm parameters and testosterone levels: a meta-analysis. Diabetol Metab Syndr. 2021; 13(1): 109.
2. Carvalho MG, Silva KM, Aristizabal VHV, Ortiz PEO, Paranzini CS, Melchert A, et al. Effects of obesity and diabetes on sperm cell proteomics in rats. J Proteome Res. 2021; 20(5): 2628-2642.
3. Leisegang K, Sengupta P, Agarwal A, Henkel R. Obesity and male infertility: mechanisms and management. Andrologia.
2021; 53(1): e13617.
4. Sexton WJ, Jarow JP. Effect of diabetes mellitus upon male reproductive function. Urology. 1997; 49(4): 508-513.
5. Chan JL, Mantzoros CS. Leptin and the hypothalamic-pituitary regulation of the gonadotropin-gonadal axis. Pituitary. 2001; 4(1-2): 87-92.
6. Pitteloud N, Hardin M, Dwyer AA, Valassi E, Yialamas M, Elahi D, et al. Increasing insulin resistance is associated with a decrease in Leydig cell testosterone secretion in men. J Clin Endocrinol Metab. 2005; 90(5): 2636-2641.
7. Condorelli RA, La Vignera S, Mongioì LM, Alamo A, Calogero AE. Diabetes mellitus and infertility: different pathophysiological effects in type 1 and type 2 on sperm function. Front Endocrinol (Lausanne). 2018; 9: 268.
8. Abbasihormozi SH, Babapour V, Kouhkan A, Niasari Naslji A, Afraz K, Zolfaghary Z, et al. Stress hormone and oxidative
stress biomarkers link obesity and diabetes with reduced fertility potential. Cell J. 2019; 21(3): 307-313.
9. Mousavi MS, Shahverdi A, Drevet J, Akbarinejad V, Esmaeili V, Sayahpour FA, et al. Peroxisome proliferator-activated receptors (PPARs) levels in spermatozoa of normozoospermic and asthenozoospermic men. Syst Biol Reprod Med. 2019; 65(6): 409-419.
10. Naeini Z, Toupchian O, Vatannejad A, Sotoudeh G, Teimouri M, Ghorbani M, et al. Effects of DHA-enriched fish oil on gene expression levels of p53 and NF-κB and PPAR-γ activity in PBMCs of patients with T2DM: a randomized, double-blind, clinical trial. Nutr Metab Cardiovasc Dis. 2020; 30(3): 441-447.
11. Serasanambati M, Chilakapati SR. Function of nuclear factor kappa B (NF-kB) in human diseases-a review. South Indian J Biol Sci. 2016; 2(4): 368-387.
12. El-Hoseny R, Neamatallah M, Alghobary M, Zalata A, Comhaire F, El-Beah SM. The possible role of NF-κB1 Rs28362491 polymorphism in male fertility of Egyptian population. Andrologia. 2020; 52(7): e13659.
13. Pentikäinen V, Suomalainen L, Erkkilä K, Martelin E, Parvinen M, Pentikäinen MO, et al. Nuclear factor-kappa B activation in human testicular apoptosis. Am J Pathol. 2002; 160(1): 205-218.
14. Catrysse L, van Loo G. Inflammation and the metabolic syndrome: the tissue-specific functions of NF-κB. Trends Cell Biol. 2017; 27(6): 417-429.
15. Santoro M, De Amicis F, Aquila S, Bonofiglio D. Peroxisome proliferator-activated receptor gamma expression along the male genital system and its role in male fertility. Hum Reprod. 2020; 35(9): 2072-2085.
16. Ranganathan P, Kattal N, Moustafa MH, Sharma RK, Thomas AJ, Agarwal A. Correlation of nuclear factor KAPPA B (NFΚB) with sperm quality and clinical diagnoses in infertile men. Fertil Steril. 2002; 78(1): S95.
17. Esmaeili V, Zendehdel M, Shahverdi A, Alizadeh A. Relationship between obesity-related markers, biochemical metabolic parameters, hormonal profiles and sperm parameters among men attending an infertility clinic. Andrologia. 2022; 54(1): e14524.
18. Abbasihormozi S, Kouhkan A, Alizadeh AR, Shahverdi AH, Nasr-Esfahani MH, Sadighi Gilani M A, et al. Association of vitamin D status with semen quality and reproductive hormones in Iranian subfertile men. Andrology. 2017; 5(1): 113-118.
19. Ernst O, Vayttaden SJ, Fraser IDC. Measurement of NF-κB activation in TLR-activated macrophages. Methods Mol Biol. 2018; 1714: 67-78.
20. Wang S, Sun J, Wang J, Ping Z, Liu L. Does obesity based on body mass index affect semen quality?-A meta-analysis and systematic review from the general population rather than the infertile population. Andrologia. 2021; 53(7): e14099.
21. Abdel-Fadeil MR, Abd Allah ESH, Iraqy HM, Elgamal DA, Abdel-Ghani MA. Experimental obesity and diabetes reduce male fertility: potential involvement of hypothalamic Kiss-1, pituitary nitric oxide, serum vaspin and visfatin. Pathophysiology. 2019; 26(3-4): 181-189.
22. Mohammad MHS, Ameen EM. Impact of diabetes and obesity on human fertility and semen quality. Zanco J Pure Appl Sci T. 2021; 33(1): 42-54.
23. Leisegang K, Henkel R, Agarwal A. Obesity and metabolic syndrome associated with systemic inflammation and the impact on the male reproductive system. Am J Reprod Immunol. 2019; 82(5): e13178.
24. Al-Ali MB, Gutschi T, Pummer K, Zigeuner R, Brookman-May S, Wieland WF, et al. Body mass index has no impact on sperm quality but on reproductive hormones levels. Andrologia. 2014; 46(2): 106-111.
25. Aquila S, Bonofiglio D, Gentile M, Middea E, Gabriele S, Belmonte M, et al. Peroxisome proliferator-activated receptor (PPAR) gamma is expressed by human spermatozoa: its potential role on the sperm physiology. J Cell Physiol. 2006; 209(3): 977-986.
26. Fan W, Xu Y, Liu Y, Zhang Z, Lu L, Ding Z. Obesity or overweight, a chronic inflammatory status in male reproductive system, leads to mice and human subfertility. Front Physiol. 2018; 8: 1117.
27. Özata DM, Li X, Lee L, Liu J, Warsito D, Hajeri P, et al. Loss of miR-514a-3p regulation of PEG3 activates the NF-kappa B pathway in human testicular germ cell tumors. Cell Death Dis. 2017; 8(5): e2759.
28. Singh VP, Bali A, Singh N, Jaggi AS. Advanced glycation end products and diabetic complications. Korean J Physiol Pharmacol. 2014; 18(1): 1-14.
29. Lv X, Lv GH, Dai GY, Sun HM, Xu HQ. Food-advanced glycation end products aggravate the diabetic vascular complications via modulating the AGEs/RAGE pathway. Chin J Nat Med. 2016; 14(11): 844-855.
30. Biscetti F, Rando MM, Nardella E, Cecchini AL, Pecorini G, Landolfi R, et al. High mobility group Box-1 and diabetes mellitus complications: state of the art and future perspectives. Int J Mol Sci. 2019; 20(24): 6258.
31. Li J, Jiang R, Cong X, Zhao Y. UCP2 gene polymorphisms in obesity and diabetes, and the role of UCP2 in cancer. FEBS Lett. 2019; 593(18): 2525-2534.
32. Cataldi S, Costa V, Ciccodicola A, Aprile M. PPARγ and diabetes: beyond the genome and towards personalized medicine. Curr Diab Rep. 2021; 21(6): 18.
33. Crowley CA, Smith WPW, Seah KTM, Lim SK, Khan WS. Cryopreservation of human adipose tissues and adipose-derived stem cells with DMSO and/or trehalose: a systematic review. Cells. 2021; 10(7): 1837.