Effect of Different High-Fat and Advanced Glycation End-Products Diets in Obesity and Diabetes-Prone C57BL/6 Mice on Sperm Function

Document Type : Original Article


1 Department of Animal Biotechnology, Reproductive Biomedicine Research Center, Royan Institute for Biotechnology, ACECR, Isfahan, Iran

2 1. Department of Reproductive Biotechnology, Reproductive Biomedicine Research Center, Royan Institute for Biotechnology, ACECR, Isfahan, Iran

3 Department of Animal Biotechnology, Cell Science Research Center, Royan Institute for Biotechnology, ACECR, Isfahan, Iran

4 Department of Animal Biotechnology, Reproductive Biomedicine Research Center, Royan Ins titute for Biotechnology, ACECR, Isfahan, Iran


We aimed to compare the effects of using high-fat (HF) and advanced glycation end-products (AGEs) containing dietsto induce obesity and diabetes on sperm function in mice.
Materials and Methods:
 In this experimental study, twenty-five 4-week old C57BL/6 mice were divided into 5 groups and were fed with control, 45% HF, 60% HF, 45% AGEs-HF, or 60% AGEs-HF diet. After 28 weeks, fast blood sugar, glucose intolerance, insulin concentration, homeostatic model assessments (HOMA) for insulin resistance (IR) and HOMA for beta cells (HOMA beta) from systematic blood were assessed. In addition, body weight, morphometric characteristics of testes, sperm parameters, DNA damage (AO), protamine deficiency (CMAA3), and sperm membrane (DCFH-DA) and intracellular (BODIPY) lipid peroxidation were measured.
 Body mass and fasting blood sugar increased significantly in all experimental groups compared to the control group. Insulin concentration, glucose intolerance, HOMA IR, and HOMA beta were also increased significantly with higher levels of fat and AGEs in all four diets (p <0.05). The changes in the 60% HF-AGEs group, however, were more significant (p <0.001). Morphometric characteristics of the testis, sperm concentration, and sperm morphology in the diet groups did not significantly differ from the control group, while sperm motility and DNA damage in the 45%HF were significantly low. Although for protamine deficiency, both 60% HF-AGEs and 45% HF showed a significant increase compared to the control, the mean of sperm lipid in the 45% HF group and intracellular peroxidation in the 60% HF-AGEs group had the highest and the lowest increases, respectively.
Our results, interestingly, showed that isthe negative effects of a diet containing AGEs on examined parameters are lessthan those in HF diets. One possible reason is detoxification through the activation of the protective glyoxalase pathway asthe result of the chronic AGEs increase in the body.


1. American Diabetes Association. Diagnosis and classification of diabetes mellitus diabetes care. 2012; 35 Suppl 1: S64-S71.
2. World Health Organization. Laboratory manual for the examination and processing of human semen. Cambridge: Cambridge Univ Press; 2010.
3. Bettiga A, Fiorio F, Di Marco F, Trevisani F, Romani A, Porrini E, et al. The modern wes tern diet rich in advanced glycation end-products (AGEs): an overview of its impact on obesity and early progression of renal pathology. Nutrients. 2019; 11(8): 1748.
4. Nevin C, McNeil L, Ahmed N, Murgatroyd C, Brison D, Carroll M. Investigating the glycating effects of glucose, glyoxal and methylglyoxal on human sperm. Sci Rep. 2018; 8: 9002.
5. Vlassara H, Palace M. Diabetes and advanced glycation endproducts. J Intern Med. 2002; 251(2): 87-101.
6. Niwas Jangir R, Chand Jain G. Diabetes mellitus induced impairment of male reproductive functions: a review. Curr Diabetes Rev. 2014; 10(3): 147-157.
7. Karimi J, Goodarzi M, Tavilani H, Khodadadi I, Amiri I. Relationship between advanced glycation end products and increased lipid peroxidation in semen of diabetic men. Diabetes Res Clin Pract. 2011; 91(1): 61-66.
8. Cardoso AR, Kakimoto PA, Kowaltowski AJ. Diet-sensitive sources of reactive oxygen species in liver mitochondria: role of very long chain acyl-CoA dehydrogenases. PLoS One. 2013; 8(10): e77088.
9. Nadal-Casellas A, Amengual-Cladera E, Proenza AM, Lladó I, Gianotti M. Long-term high-fat-diet feeding impairs mitochondrial biogenesis in liver of male and female rats. Cell Physiol Biochem. 2010; 26(3): 291-302.
10. Satapati S, Sunny NE, Kucejova B, Fu X, He TT, Méndez-Lucas A, et al. Elevated TCA cycle function in the pathology of diet-induced hepatic insulin resistance and fatty liver. J Lipid Resh. 2012; 53(6): 1080-1092.
11. Sandu O, Song K, Cai W, Zheng F, Uribarri J, Vlassara H. Insulin resistance and type 2 diabetes in high-fat–fed mice are linked to high glycotoxin intake. Diabetes. 2005; 54(8): 2314-2319.
12. Andrikopoulos S, Blair AR, Deluca N, Fam BC, Proietto J. Evaluating the glucose tolerance test in mice. Am J Physiol Endocrinol Metab. 2008; 295(6) :E1323- E1332.
13. Wang CY, Liao JK. A mouse model of diet-induced obesity and insulin resistance. Methods Mol Biol. 2012; 821: 421-433.
14. Sun C, Li X, Liu L, Conet M, Guan Y, Fan Y, et al. Effect of fasting time on measuring mouse blood glucose level. Int J Clin Exp med. 2016; 9(2): 4186-4189.
15. Wallace TM, Levy JC, Matthews DR. Use and abuse of HOMA modeling. Diabetes Care. 2004; 27(6): 1487-1495.
16. Matthews DR, Hosker JP, Rudenski AS, Naylor BA, Treacher DF, Turner RC. Homeostasis model assessment: insulin resistance and beta-cell function from fasting plasma glucose and insulin concentrations in man. Diabetologia. 1985; 28(7): 412-419.
17. Afiyani AA, Deemeh MR, Tavalaee M, Razi M, Bahadorani M, Shokrollahi B, et al. Evaluation of heat-shock protein A2 (HSPA2) in male rats before and after varicocele induction. Mol Reprod Dev. 2014; 81(8): 766-776.
18. Aitken RJ, Wingate JK, De Iuliis GN, McLaughlin EA. Analysis of lipid peroxidation in human spermatozoa using BODIPY C11. Mol Hum Reprod. 2007; 13(4): 203-211.
19. Kiani-Esfahani A, Tavalaee M, Deemeh MR, Hamiditabar M, Nasr-Esfahani MH. DHR123: an alternative probe for assessment of ROS in human spermatozoa. Sys t Biol Reprod Med. 2012; 58(3): 168-174.
20. de Assis AM, Rieger DK, Longoni A, Battu C, Raymundi S, da Rocha RF, et al. High fat and highly thermolyzed fat diets promote insulin resistance and increase DNA damage in rats. Exp Biol Med (Maywood). 2009; 234(11): 1296-1304.
21. Karimi J, Goodarzi M, Tavilani H, Khodadadi I, Amiri I. Increased receptor for advanced glycation end products in spermatozoa of diabetic men and its association with sperm nuclear DNA fragmentation. Andrologia. 2012; 44 Suppl 1: 280-286.
22. Mallidis C, Agbaje IM, Rogers DA, Glenn JV, Pringle R, Atkinson AB, et al. Advanced glycation end products accumulate in the reproductive tract of men with diabetes. Int J Androl. 2009; 32(4): 295-305.
23. Mallidis C, Agbaje I, Rogers D, Glenn J, McCullough S, Atkinson AB, et al. Distribution of the receptor for advanced glycation end products in the human male reproductive tract: prevalence in men with diabetes mellitus. Human Reprod. 2007; 22(8): 2169-2177.
24. Ferramosca A, Moscatelli N, Di Giacomo M, Zara V. Dietary fatty acids influence sperm quality and function. Andrology. 2017; 5(3): 423-430.
25. Lu YC, Sudirman S, Mao CF, Kong ZL. Glycoprotein from Mytilus edulis extract inhibits lipid accumulation and improves male reproductive dysfunction in high-fat diet-induced obese rats. Biomed Pharmacother. 2019; 109: 369-376.
26. Ferramosca A, Conte A, Moscatelli N, Zara V. A high-fat diet negatively affects rat sperm mitochondrial respiration. Andrology. 2016; 4(3): 520-525.
27. Portela JM, Tavares RS, Mota PC, Ramalho-Santos J, Amaral S. High glucose concentrations per se do not adversely affect human sperm function in vitro. Reproduction. 2015; 150(1): 77-84.
28. Chen MC, Lin JA, Lin HT, Chen SY, Yen GC. Potential effect of advanced glycation end products (AGEs) on spermatogenesis and sperm quality in rodents. Food Funct. 2019; 10(6): 3324-3333.
29. Chen SJ, Aikawa C, Yoshida R, Matsui T. Methylglyoxal-derived hydroimidazolone residue of plasma protein can behave as a predictor of prediabetes in Spontaneously Diabetic Torii rats. Physiol Rep. 2015; 3(8): e12477.
30. Antognelli C, Mancuso F, Frosini R, Arato I, Calvitti M, Calafiore R, et al. Testosterone and follicle stimulating hormone-dependent glyoxalase 1 up-regulation sus tains the viability of porcine sertoli cells through the control of hydroimidazolone- and argpyrimidine-mediated NF-κB pathway. Am J Pathol. 2018; 188(11): 2553-2563.
31. Jain M, Nagar P, Sharma A, Batth R, Aggarwal S, Kumari S, et al. GLYI and D-LDH play key role in methylglyoxal detoxification and abiotic s tress tolerance. Sci Rep. 2018; 8(1): 5451.
32. Rodrigues T, Matafome P, Sereno J, Almeida J, Castelhano J, Gamas L, et al. Methylglyoxal-induced glycation changes adipose tissue vascular architecture, flow and expansion, leading to insulin resistance. Sci Rep. 2017; 7(1): 1698.
33. Norouzirad R, González-Muniesa P, Ghasemi A. Hypoxia in obesity and diabetes: potential therapeutic effects of hyperoxia and nitrate. Oxid Med Cell Longev. 2017; 2017: 5350267.
34. Turkseven S, Ertuna E, Yetik-Anacak G, Yasa M. Methylglyoxal causes endothelial dysfunction: the role of endothelial nitric oxide synthase and AMP-activated protein kinase α. J Basic Clin Physiol Pharmacol. 2014; 25(1): 109-115.
35. Reyes JG, Farias JG, Henríquez-Olavarrieta S, Madrid E, Parraga M, Zepeda AB, et al. The hypoxic tes ticle: physiology and pathophysiology. Oxid Med Cell Longev. 2012; 2012: 929285 .
36. Aitken RJ. DNA damage in human spermatozoa; important contributor to mutagenesis in the offspring. Transl Androl Urol. 2017; 6 Suppl 4: S761- S764.
37. Chen Y, Wu Y, Gan X, Liu K, Lv X, Shen H, et al. Iridoid glycoside from Cornus officinalis ameliorated diabetes mellitus-induced testicular damage in male rats: Involvement of suppression of the AGEs/RAGE/p38 MAPK signaling pathway. J Ethnopharmacol. 2016; 194: 850-860.
38. Charalampidou S, Simitsopoulou Μ, Skoura L, Tziomalos K, Koulourida V, Goulis DG. Soluble receptor for advanced glycation end products in male infertility. Hippokratia. 2017; 21(1): 19-24.
39. Hu X, Ge X, Liang W, Shao Y, Jing J, Wang C, et al. Effects of saturated palmitic acid and omega-3 polyunsaturated fatty acids on Sertoli cell apoptosis. Sys t Biol Reprod Med. 2018; 64(5): 368-380.
40. Schwarzenbolz U, Mende S, Henle T. Model s tudies on protein glycation: influence of cys teine on the reactivity of arginine and lysine residues toward glyoxal. Ann N Y Acad Sci. 2008; 1126: 248-252.