Lactobacillus plantarum and Lactobacillus helveticus modulate SIRT1, Caspase3 and Bcl-2 in the testes of high-fructose-fed rats

Abstract

Background and Aims: The influence of a high-fructose diet and probiotics on the male reproductive system and the testicular apoptotic pathway has been poorly documented. In this study, we aimed to investigate the influence of Lactobacillus plantarum and Lactobacillus helveticus supplementation on apoptotic factors such as sirtuin1, caspase3 and bcl-2 on the testicular tissue of high-fructose-fed rats. Methods: Fructose was given to the rats as a 20% solution in drinking water for 15 weeks. Gene expressions were established by real-time PCR. Protein levels were determined by Western blot analysis. Results: Fructose consumption did not change mRNA expression of SIRT1, but did resulted in a decreased protein level. Dietary fructose reduced bcl-2 mRNA and protein expressions, whereas no changes were observed in the gene and protein expression levels of factor caspase-3. Both Lactobacillus supplementations increased SIRT1 protein expression without changing the mRNA levels in fructose-fed rats. The supplementations with both probiotics produced a significant downregulation on caspase3 mRNA and protein levels. Bcl-2 proetin level increases with both probiotics supplementation while, mRNA level did not show difference in L.plantarum, but increased in L. helveticus supplementation. Conclusion: Treatments with L.plantarum and L.helveticus can reduce testicular apoptosis induced by dietary high-fructose in rats via suppressing caspase3 and promoting sirt1 and bcl-2 protein expressions.

Keywords

• Dietary fructose
• Lactobacillus plantarum
• Lactobacillus helveticus
• apoptosis
• testes

References

1. • Akar, F., Uludag, O., Aydin, A., Aytekin, Y. A., Elbeg, S., Tuzcu, M., & Sahin, K. (2012). High-fructose corn syrup causes vascular dysfunction associated with metabolic disturbance in rats: protective effect of resveratrol. Food and Chemical Toxicology, 50(6), 2135–2141.
2. • Alsemeh, A. E., Samak, M. A., & El-Fatah, S. S. A. (2020). Therapeutic prospects of hydroxytyrosol on experimentally induced diabetic testicular damage: potential interplay with AMPK expression. Cell and Tissue Research, 380(1), 173–189.
3. • Babacanoglu, C., Yildirim, N., Sadi, G., Pektas, M. B., & Akar, F. (2013). Resveratrol prevents high-fructose corn syrup-induced vascular insulin resistance and dysfunction in rats. Food and Chemical Toxicology, 60, 160–167.
4. • Backhed, F., Ley, R. E., Sonnenburg, J. L., Peterson, D. A., & Gordon, J. I. (2005). Host-bacterial mutualism in the human intestine. Science, 307(5717), 1915–1920.
5. • Balakumar, M., Raji, L., Prabhu, D., Sathishkumar, C., Prabu, P., Mohan, V., & Balasubramanyam, M. (2016). High-fructose diet is as detrimental as high-fat diet in the induction of insulin resistance and diabetes mediated by hepatic/pancreatic endoplasmic reticulum (ER) stress. Molecular and Cellular Biochemistry, 423(1-2), 93–104.
6. • Bouhafs, L., Moudilou, E. N., Exbrayat, J. M., Lahouel, M., & Idoui, T. (2015). Protective effects of probiotic Lactobacillus plantarum BJ0021 on liver and kidney oxidative stress and apoptosis induced by endosulfan in pregnant rats. Renal Failure, 37(8), 1370–1378.
7. • Brady, H. J. M., & Gil-Gomez, G. (1998). Molecules in focus - Bax. The pro-apoptotic Bcl-2 family member, Bax. International Journal of Biochemistry and Cell Biology, 30(6), 647–650.
8. • Caldas, A. D., Porto, A. L., Motta, L. D., & Casulari, L. A. (2009). Relationship between insulin and hypogonadism in men with metabolic syndrome. Arquivos Brasileiros de Endocrinologia e Metabologia, 53(8), 1005–1011.

9. • Chaki, S. P., Misro, M. M., Gautam, D. K., Kaushik, M., Ghosh, D., & Chainy, G. B. (2006). Estradiol treatment induces testicular oxidative stress and germ cell apoptosis in rats. Apoptosis, 11(8), 1427– 1437.
10. • Choi, Y., Abdelmegeed, M. A., & Song, B. J. (2017). Diet high in fructose promotes liver steatosis and hepatocyte apoptosis in C57BL/6J female mice: Role of disturbed lipid homeostasis and increased oxidative stress. Food and Chemical Toxicology, 103, 111–121.
11. • Cory, S., Huang, D. C., & Adams, J. M. (2003). The Bcl-2 family: roles in cell survival and oncogenesis. Oncogene, 22(53), 8590–8607.
12. • Coussens, M., Maresh, J. G., Yanagimachi, R., Maeda, G., & Allsopp, R. (2008). Sirt1 deficiency attenuates spermatogenesis and germ cell function. PLoS One, 3(2), e1571.
13. • Cregan, S. P., Dawson, V. L., & Slack, R. S. (2004). Role of AIF in caspase-dependent and caspase-independent cell death. Oncogene, 23(16), 2785–2796.
14. • Dandona, P., & Dhindsa, S. (2011). Update: Hypogonadotropic hypogonadism in type 2 diabetes and obesity. Journal of Clinical Endocrinology and Metabolism, 96(9), 2643–2651.
15. • Dhindsa, S., Miller, M. G., McWhirter, C. L., Mager, D. E., Ghanim, H., Chaudhuri, A., & Dandona, P. (2010). Testosterone concentrations in diabetic and nondiabetic obese men. Diabetes Care, 33(6), 1186–1192.
16. • Du, Z., Qiu, Z., Wang, Z., & Wang, X. (2018). The inhibitory effects of soybean isoflavones on testicular cell apoptosis in mice with type 2 diabetes. Experimental and Therapeutic Medicine, 15(1), 305–309.
17. • Ebrahimi, F., Schuetz, P., Mueller, B., Urwyler, S. A., Donath, M. Y., & Christ-Crain, M. (2017). Effects of IL-1 [beta] on the hypothalamic-pituitary- gonadal axis in men with obesity and metabolic syndrome-A randomized, double-blind, placebo-controlled trial. Paper presented at the 19th European Congress of Endocrinology. Endocrine Abstracts (2017) 49 EP687.
18. • Faid, I., Al-Hussaini, H., & Kilarkaje, N. (2015). Resveratrol alleviates diabetes-induced testicular dysfunction by inhibiting oxidative stress and c-Jun N-terminal kinase signaling in rats. Toxicology and Applied Pharmacology, 289(3), 482–494.
19. • Girard, S. A., Bah, T. M., Kaloustian, S., Lada-Moldovan, L., Rondeau, I., Tompkins, T. A., . . . Rousseau, G. (2009). Lactobacillus helveticus and Bifidobacterium longum taken in combination reduce the apoptosis propensity in the limbic system after myocardial infarction in a rat model. British Journal of Nutrition, 102(10), 1420–1425.
20. • Green, D. R., & Llambi, F. (2015). Cell Death Signaling. Cold Spring Harbor Perspectives in Biology, 7(12), a006080.
21. • Hayashi, K., Kojima, R., & Ito, M. (2006). Strain differences in the diabetogenic activity of streptozotocin in mice. Biological and Pharmaceutical Bulletin, 29(6), 1110–1119.
22. • Hendijani, F., & Akbari, V. (2018). Probiotic supplementation for management of cardiovascular risk factors in adults with type II diabetes: A systematic review and meta-analysis. Clinical Nutrition, 37(2), 532–541.
23. • Honda, K., Moto, M., Uchida, N., He, F., & Hashizume, N. (2012). Antidiabetic effects of lactic acid bacteria in normal and type 2 diabetic mice. Journal of Clinical Biochemistry and Nutrition, 51(2), 96–101.
24. • Hotel, A. C. P., & Cordoba, A. (2001). Health and nutritional properties of probiotics in food including powder milk with live lactic acid bacteria. Prevention, 5(1), 1–10.
25. • Hu, Q. H., Zhang, X., Pan, Y., Li, Y. C., & Kong, L. D. (2012). Allopurinol, quercetin and rutin ameliorate renal NLRP3 inflammasome activation and lipid accumulation in fructose-fed rats. Biochemical Pharmacology, 84(1), 113–125.
26. • Huang, L., Zhao, Z., Duan, C., Wang, C., Zhao, Y., Yang, G., . . . Li, S. (2019). Lactobacillus plantarum C88 protects against aflatoxin B 1-induced liver injury in mice via inhibition of NF-κB–mediated inflammatory responses and excessive apoptosis. BMC Microbiology, 19(1), 170.
27. • Koh, P. O. (2007a). Streptozotocin-induced diabetes increases apoptosis through JNK phosphorylation and Bax activation in rat testes. Journal of Veterinary Medical Science, 69(9), 969–971.
28. • Koh, P. O. (2007b). Streptozotocin-induced diabetes increases the interaction of Bad/Bcl-XL and decreases the binding of pBad/14- 3-3 in rat testis. Life Sciences, 81(13), 1079–1084.
29. • Korkmaz, O. A., Sadi, G., Kocabas, A., Yildirim, O. G., Sumlu, E., Koca, H. B., . . . Akar, F. (2019a). Lactobacillus helveticus and Lactobacillus plantarum modulate renal antioxidant status in a rat model of fructose-induced metabolic syndrome. Archives of Biological Sciences, 71(2), 265–273.
30. • Korkmaz, O. A., Sumlu, E., Koca, H. B., Pektas, M. B., Kocabas, A., Sadi, G., & Akar, F. (2019b). Effects of Lactobacillus Plantarum and Lactobacillus Helveticus on Renal Insulin Signaling, Inflammatory Markers, and Glucose Transporters in High-Fructose-Fed Rats. Medicina (Kaunas, Lithuania), 55(5), 207.
31. • Kwon, J., Kim, B., Lee, C., Joung, H., Kim, B.-K., Choi, I. S., & Hyun, C.- K. (2020). Comprehensive amelioration of high-fat diet-induced metabolic dysfunctions through activation of the PGC-1α pathway by probiotics treatment in mice. PLoS One, 15(2), e0228932.
32. • Ley, R. E., Backhed, F., Turnbaugh, P., Lozupone, C. A., Knight, R. D., & Gordon, J. I. (2005). Obesity alters gut microbial ecology. Proceedings of the National Academy of Sciences of the United States of America, 102(31), 11070–11075.
33. • Liu, H., Zhang, S., Liu, C., Wu, J., Wang, Y., Yuan, L., . . . Zhuang, D. (2018). Resveratrol ameliorates microcystin-LR-induced testis germ cell apoptosis in rats via SIRT1 signaling pathway activation. Toxins, 10(6), 235.
34. • Liu, Q., Liu, Y., Li, F., Gu, Z., Liu, M., Shao, T., . . . Feng, W. (2020). Probiotic culture supernatant improves metabolic function through FGF21-adiponectin pathway in mice. The Journal of Nutritional Biochemistry, 75, 108256.
35. • Lowry, O. H., Rosebrough, N. J., Farr, A. L., & Randall, R. J. (1951). Protein measurement with the Folin phenol reagent. Journal of Biological Chemistry, 193(1), 265–275.
36. • Lu, X. L., Zhao, C. H., Yao, X. L., & Zhang, H. (2017). Quercetin attenuates high fructose feeding-induced atherosclerosis by suppressing inflammation and apoptosis via ROS-regulated PI3K/AKT signaling pathway. Biomedicine and Pharmacotherapy, 85, 658–671.
37. • Markowiak, P., & Slizewska, K. (2017). Effects of Probiotics, Prebiotics, and Synbiotics on Human Health. Nutrients, 9(9), 1021.
38. • McBurney, M. W., Yang, X., Jardine, K., Hixon, M., Boekelheide, K., Webb, J. R., . . . Lemieux, M. (2003). The mammalian SIR2α protein has a role in embryogenesis and gametogenesis. Molecular and cellular biology, 23(1), 38–54.
39. • Meydanli, E. G., Gumusel, A., Ozkan, S., Tanriverdi, G., Balci, M. B. C., Develi Is, S., . . . Bekpinar, S. (2018). Effects of resveratrol on highfructose- induced testis injury in rats. Ultrastructural Pathology, 42(1), 65–73.
40. • Mohammadi, G., Dargahi, L., Naserpour, T., Mirzanejad, Y., Alizadeh, S. A., Peymani, A., & Nassiri-Asl, M. (2019). Probiotic mixture of Lactobacillus helveticus R0052 and Bifidobacterium longum R0175 attenuates hippocampal apoptosis induced by lipopolysaccharide in rats. International Microbiology, 22(3), 317–323.
41. • Morrison, C. D., & Brannigan, R. E. (2015). Metabolic syndrome and infertility in men. Best Practice & Research: Clinical Obstetrics & Gynaecology, 29(4), 507–515. • Nna, V. U., Bakar, A. B. A., Ahmad, A., & Mohamed, M. (2018). Diabetes- induced testicular oxidative stress, inflammation, and caspase-dependent apoptosis: the protective role of metformin. Archives of Physiology and Biochemistry, 1–12.
42. • Park, D. Y., Ahn, Y. T., Huh, C. S., McGregor, R. A., & Choi, M. S. (2013). Dual probiotic strains suppress high fructose-induced metabolic syndrome. World Journal of Gastroenterology, 19(2), 274–283.
43. • Passos, J. F., Saretzki, G., Ahmed, S., Nelson, G., Richter, T., Peters, H., . . . von Zglinicki, T. (2007). Mitochondrial dysfunction accounts for the stochastic heterogeneity in telomere-dependent senescence. PLoS Biology, 5(5), e110.
44. • Pektas, M. B., Koca, H. B., Sadi, G., & Akar, F. (2016). Dietary Fructose Activates Insulin Signaling and Inflammation in Adipose Tissue: Modulatory Role of Resveratrol. BioMed Research International, 2016, 8014252.
45. • Pektas, M. B., Sadi, G., & Akar, F. (2015). Long-Term Dietary Fructose Causes Gender-Different Metabolic and Vascular Dysfunction in Rats: Modulatory Effects of Resveratrol. Cellular Physiology and Biochemistry, 37(4), 1407–1420.
46. • Pektas, M. B., Yucel, G., Koca, H. B., Sadi, G., Yildirim, O. G., Ozturk, G., & Akar, F. (2017). Dietary Fructose-Induced Hepatic Injury in Male and Female Rats: Influence of Resveratrol. Drug Research (Stuttg), 67(2), 103–110.
47. • Plaza-Diaz, J., Ruiz-Ojeda, F. J., Vilchez-Padial, L. M., & Gil, A. (2017). Evidence of the Anti-Inflammatory Effects of Probiotics and Synbiotics in Intestinal Chronic Diseases. Nutrients, 9(6), 555.
48. • Rastrelli, G., Filippi, S., Sforza, A., Maggi, M., & Corona, G. (2018). Metabolic syndrome in male hypogonadism. In Metabolic Syndrome Consequent to Endocrine Disorders (Vol. 49, pp. 131–155): Karger Publishers.
49. • Rosa, D. D., Grzeskowiak, L. M., Ferreira, C. L., Fonseca, A. C., Reis, S. A., Dias, M. M., . . . Peluzio Mdo, C. (2016). Kefir reduces insulin resistance and inflammatory cytokine expression in an animal model of metabolic syndrome. Food & Function, 7(8), 3390–3401.
50. • Sadi, G., Ergin, V., Yilmaz, G., Pektas, M. B., Yildirim, O. G., Menevse, A., & Akar, F. (2015). High-fructose corn syrup-induced hepatic dysfunction in rats: improving effect of resveratrol. European Journal of Nutrition, 54(6), 895–904.
51. • Shahin, S., Singh, S. P., & Chaturvedi, C. M. (2018). 2.45 GHz microwave radiation induced oxidative and nitrosative stress mediated testicular apoptosis: Involvement of a p53 dependent bax-caspase- 3 mediated pathway. Environmental Toxicology, 33(9), 931–945.
52. • Simoes, V. L., Alves, M. G., Martins, A. D., Dias, T. R., Rato, L., Socorro, S., & Oliveira, P. F. (2013). Regulation of apoptotic signaling pathways by 5alpha-dihydrotestosterone and 17beta-estradiol in immature rat Sertoli cells. Journal of Steroid Biochemistry and Molecular Biology, 135, 15–23.
53. • Song, L., Chen, T. Y., Zhao, X. J., Xu, Q., Jiao, R. Q., Li, J. M., & Kong, L. D. (2019). Pterostilbene prevents hepatocyte epithelial-mesenchymal transition in fructose-induced liver fibrosis through suppressing miR-34a/Sirt1/p53 and TGF-β1/Smads signalling. British journal of pharmacology, 176(11), 1619–1634.
54. • Song, W., Liu, M. G., Zhang, J. B., Zhang, J. J., Sun, M. M., & Yu, Q. K. (2016). Mechanism of action of EBV, Bcl-2, p53, c-Myc and Rb in non-Hodgkin’s lymphoma. European Review for Medical and Pharmacological Sciences, 20(6), 1093–1097.
55. • Sumlu, E., Bostanci, A., Sadi, G., Alcigir, M. E., & Akar, F. (2020). Lactobacillus plantarum improves lipogenesis and IRS-1/AKT/eNOS signalling pathway in the liver of high-fructose-fed rats. Archives of Physiology and Biochemistry, 1–9.
56. • Tsai, E. C., Matsumoto, A. M., Fujimoto, W. Y., & Boyko, E. J. (2004). Association of bioavailable, free, and total testosterone with insulin resistance: influence of sex hormone-binding globulin and body fat. Diabetes Care, 27(4), 861–868.
57. • Tsounapi, P., Saito, M., Dimitriadis, F., Koukos, S., Shimizu, S., Satoh, K., . . . Sofikitis, N. (2012). Antioxidant treatment with edaravone or taurine ameliorates diabetes-induced testicular dysfunction in the rat. Molecular and Cellular Biochemistry, 369(1-2), 195–204.
58. • Vachharajani, V. T., Liu, T., Wang, X., Hoth, J. J., Yoza, B. K., & McCall, C. E. (2016). Sirtuins link inflammation and metabolism. Journal of Immunology Research, 8167273.
59. • Wang, H. F., Tseng, C. Y., Chang, M. H., Lin, J. A., Tsai, F. J., Tsai, C. H., . . . Tsai, C. C. (2012). Anti-inflammatory effects of probiotic Lactobacillus paracasi on ventricles of BALB/C mice treated with ovalbumin. Chinese Journal of Physiology, 55(1), 37–46.
60. • Wang, X.-l., Wu, L.-y., Zhao, L., Sun, L.-n., Liu, H.-y., Liu, G., & Guan, G.-j. (2016). SIRT1 activator ameliorates the renal tubular injury induced by hyperglycemia in vivo and in vitro via inhibiting apoptosis. Biomedicine & Pharmacotherapy, 83, 41–50.
61. • Wang, Y., Wu, Y., Wang, Y., Xu, H., Mei, X., Yu, D., . . . Li, W. (2017). Antioxidant Properties of Probiotic Bacteria. Nutrients, 9(5), 521.
62. • Xu, Y., Lei, H., Guan, R., Gao, Z., Li, H., Wang, L., . . . Xin, Z. (2014). Studies on the mechanism of testicular dysfunction in the early stage of a streptozotocin induced diabetic rat model. Biochemical and Biophysical Research Communications, 450(1), 87–92.
63. • Yadav, H., Jain, S., & Sinha, P. R. (2007). Antidiabetic effect of probiotic dahi containing Lactobacillus acidophilus and Lactobacillus casei in high fructose fed rats. Nutrition, 23(1), 62–68.
64. • Yildirim, O. G., Sumlu, E., Aslan, E., Koca, H. B., Pektas, M. B., Sadi, G., & Akar, F. (2019). High-fructose in drinking water initiates activation of inflammatory cytokines and testicular degeneration in rat. Toxicology Mechanisms and Methods, 29(3), 224–232.
65. • Zhao, L., Gu, Q., Xiang, L., Dong, X., Li, H., Ni, J., . . . Chen, G. (2017). Curcumin inhibits apoptosis by modulating Bax/Bcl-2 expression and alleviates oxidative stress in testes of streptozotocin-induced diabetic rats. Therapeutics and Clinical Risk Management, 13, 1099– 1105.
66. • Zhao, Y., Tan, Y., Dai, J., Li, B., Guo, L., Cui, J., . . . Cai, L. (2011). Exacerbation of diabetes-induced testicular apoptosis by zinc deficiency is most likely associated with oxidative stress, p38 MAPK activation, and p53 activation in mice. Toxicology Letters, 200(1-2), 100–106.
67. • Zhao, Y., Tan, Y., Dai, J., Wang, B., Li, B., Guo, L., . . . Cai, L. (2012). Zinc deficiency exacerbates diabetic down-regulation of Akt expression and function in the testis: essential roles of PTEN, PTP1B and TRB3. Journal of Nutritional Biochemistry, 23(8), 1018–1026.