Caspian Journal of Veterinary Sciences

Caspian Journal of Veterinary Sciences

Indole-3-propionic acid from the gut can mitigate leydig cell dysfunction caused by hyperglycemia by inducing antioxidant pathways

Document Type : Original Article

Authors
Sadegh Jozaie 1
Mohammad Reza Tabandeh 2
1 Department of Basic Sciences, Faculty of Veterinary Medicine, Amol University of Special Modern Technologies, Amol, Iran.
2 Department of Biochemistry and Molecular Biology, Faculty of Veterinary Medicine, Shahid Chamran University of Ahvaz, Ahvaz, Iran; Stem Cells and Transgenic Technology Research Center, Shahid Chamran University of Ahvaz, Ahvaz, Iran.
10.22034/cjvs.2026.546770.1047
Abstract
Diabetes mellitus (DM) induces hyperglycemia, increasing reactive oxygen species (ROS) and impairing leydig cell function, thereby contributing to reduced male fertility. Indole-3-propionic acid (IPA), a gut microbiota–derived tryptophan metabolite with antioxidant and anti-inflammatory properties, has not been sufficiently investigated in hyperglycemia-induced reproductive damage. This study evaluated the protective effects of IPA on TM3 leydig cells exposed to high glucose (30 mM). Cells received 20 µM IPA or 1 mM N-acetylcysteine (NAC) as a positive control. Oxidative stress markers (GSH, GSSG, GPx, GR) and pro-inflammatory cytokines (IL-1β, TNF-α) were measured. High-glucose exposure significantly reduced GSH, GPx, and GR levels while increasing GSSG, IL-1β, and TNF-α. IPA treatment increased GSH levels and GR activity and reduced TNF-α compared with the HG group, whereas GPx and IL-1β showed no significant changes. Overall, these findings indicated that IPA mitigates hyperglycemia-induced oxidative stress and inflammation mainly through enhancing the glutathione redox cycle and modulating inflammatory mediators, highlighting its potential to preserve leydig cell function.
Keywords
Hyperglycemia
Indole-3-propionic acid
Leydig cells
Male infertility, Oxidative stress
Subjects
Abildgaard, A; Elfving, B; Hokland, M; Wegener, G and Lund, S (2018). The microbial metabolite indole-3-propionic acid improves glucose metabolism in rats, but does not affect behaviour. Arch. Physiol. Biochem., 124: 306–312.
Agarwal, A; Parekh, N; Selvam, MKP; Henkel, R; Shah, R; Homa, ST; Ramasamy, R; Ko, E; Tremellen, K and Esteves, S (2019). Male oxidative stress infertility (MOSI): proposed terminology and clinical practice guidelines for management of idiopathic male infertility. World J. Men’s Health, 37: 296–312.
Agarwal, A; Virk, G; Ong, C and Du Plessis, SS (2014). Effect of oxidative stress on male reproduction. World J. Men’s Health, 32: 1.
Aggarwal, A; Misro, MM; Maheshwari, A; Sehgal, N and Nandan, D (2010). N‐acetylcysteine counteracts oxidative stress and prevents hCG‐induced apoptosis in rat Leydig cells through down regulation of caspase‐8 and JNK. Mol. Reprod. Develop., 77: 900–909.
Agus, A; Planchais, J and Sokol, H (2018). Gut microbiota regulation of tryptophan metabolism in health and disease. Cell Host Microbe, 23: 716–724.
Ahmed, RG (2005). The physiological and biochemical effects of diabetes on the balance between oxidative stress and antioxidant defense system. Med. J. Islamic World Acad. Sci., 15: 31–42.
Aitken, RJ; Smith, TB; Jobling, MS; Baker, MA and De Iuliis, GN (2014). Oxidative stress and male reproductive health. Asian J. Androl., 16: 31–38.
Akingbemi, BT (2005). Estrogen regulation of testicular function. Reprod. Biol. Endocrinol., 3: 51.
Alisik, M; Neselioglu, S and Erel, O (2019). A colorimetric method to measure oxidized, reduced and total glutathione levels in erythrocytes. J. Lab. Med., 43: 269–277.
Arora, A; Behl, T; Sehgal, A; Singh, S; Sharma, N; Bhatia, S; Sobarzo-Sanchez, E and Bungau, S (2021). Unravelling the involvement of gut microbiota in type 2 diabetes mellitus. Life Sci., 273: 119311.
Bener, A; Al-Ansari, AA; Zirie, M and Al-Hamaq, AOAA (2009). Is male fertility associated with type 2 diabetes mellitus? Int. Urol. Nephrol., 41: 777–784.
Ceriello, A (2006). Oxidative stress and diabetesassociated complications. Endocrine Prac., 12: 60–62.
Clark, BJ and Stocco, DM (2014). Cholesterol transporters of the START domain protein family in health and disease. Springer.
Defeudis, G; Mazzilli, R; Tenuta, M; Rossini, G; Zamponi, V; Olana, S; Faggiano, A; Pozzilli, P; Isidori, AM and Gianfrilli, D (2022). Erectile dysfunction and diabetes: a melting pot of circumstances and treatments. Diabetes Metabolism Res. Rev., 38: e3494.
Ding, GL; Liu, Y; Liu, ME; Pan, JX; Guo, MX; Sheng, JZ and Huang, HF (2015). The effects of diabetes on male fertility and epigenetic regulation during spermatogenesis. Asian J. Androl., 17: 948–953.
Forbes, JM and Cooper, ME (2013). Mechanisms of diabetic complications. Physiol. Rev., 93: 137–188.
Forman, HJ; Zhang, H and Rinna, A (2009). Glutathione: overview of its protective roles, measurement, and biosynthesis. Mol. Aspects Med., 30: 1–12.
Garcez, ML; Tan, VX; Heng, B and Guillemin, GJ (2020). Sodium butyrate and indole-3-propionic acid prevent the increase of cytokines and kynurenine levels in LPS-induced human primary astrocytes. Int. J. Tryptophan Res., 13: 1178646920978404.
Gesper, M; Nonnast, ABH; Kumowski, N; Stoehr, R; Schuett, K; Marx, N and Kappel, BA (2021). Gutderived metabolite indole-3-propionic acid modulates mitochondrial function in cardiomyocytes and alters cardiac function. Front. Med., 8: 648259.
Han, Z; Fu, J; Gong, A and Ren, W (2025). Bacterial indole-3-propionic acid inhibits macrophage IL-1β production through targeting methionine metabolism. Sci. China Life Sci., 68: 1118–1131.
Imani, M; Talebi, AR; Fesahat, F; Rahiminia, T; Seifati, SM and Dehghanpour, F (2021). Sperm parameters, DNA integrity, and protamine expression in patients with type II diabetes mellitus. J. Obstetr. Gynaecol., 41: 439–446.
Laleethambika, N; Anila, V; Manojkumar, C; Muruganandam, I; Giridharan, B; Ravimanickam, T and Balachandar, V (2019). Diabetes and sperm DNA damage: Efficacy of antioxidants. SN Comp. Clin. Med., 1: 49–59.
Lee-Sarwar, KA; Lasky-Su, J; Kelly, RS; Litonjua, AA and Weiss, ST (2020). Metabolome–microbiome crosstalk and human disease. Metabolites, 10: 181.
Leisegang, K (2022). Oxidative stress in men with obesity, metabolic syndrome and type 2 diabetes mellitus: Mechanisms and management of reproductive dysfunction. In Oxidative Stress and Toxicity in Reproductive Biology and Medicine: A Comprehensive Update on Male Infertility-Volume One. Springer. PP: 237–256.
Leisegang, K and Henkel, R (2018). The in vitro modulation of steroidogenesis by inflammatory cytokines and insulin in TM3 Leydig cells. Reprod. Biol. Endocrinol., 16: 26.
Leylabadlo, HE; Sanaie, S; Heravi, FS; Ahmadian, Z and Ghotaslou, R (2020). From role of gut microbiota to microbial-based therapies in type 2-diabetes. Infect. Gen. Evol., 81: 104268.
Menni, C; Hernandez, MM; Vital, M; Mohney, RP; Spector, TD and Valdes, AM (2019). Circulating levels of the anti-oxidant indoleproprionic acid are associated with higher gut microbiome diversity. Gut Microbes, 10: 688–695.
Mohajeri, MH; Brummer, RJM; Rastall, RA; Weersma, RK; Harmsen, HJM; Faas, M and Eggersdorfer, M (2018). The role of the microbiome for human health: from basic science to clinical applications. Eur. J. Nutr., 57: 1–14.
Negatu, DA; Gengenbacher, M; Dartois, V and Dick, T (2020). Indole propionic acid, an unusual antibiotic produced by the gut microbiota, with antiinflammatory and antioxidant properties. Front. Microbiol., 11: 575586.
Nishikawa, T; Edelstein, D; Du, XL; Yamagishi, S; Matsumura, T; Kaneda, Y; Yorek, MA; Beebe, D; Oates, PJ and Hammes, HP (2000). Normalizing mitochondrial superoxide production blocks three pathways of hyperglycaemic damage. Nature, 404: 787–790.
Riris, AAIDC; I’tishom, R and Khaerunnisa, S (2021). Role of antioxidant to protect Leydig cells induced by reactive oxygen species: A literature review. Med. J. Faculty Med. Muhammadiyah Surabaya, 5: 49–60.
Rolo, AP and Palmeira, CM (2006). Diabetes and mitochondrial function: role of hyperglycemia and oxidative stress. Toxicol. Appl. Pharmacol., 212: 167–178.
Saeedi, P; Petersohn, I; Salpea, P; Malanda, B; Karuranga, S; Unwin, N; Colagiuri, S; Guariguata, L; Motala, AA and Ogurtsova, K (2019). Global and regional diabetes prevalence estimates for 2019 and projections for 2030 and 2045: Results from the International Diabetes Federation Diabetes Atlas. Diabetes Res. Clin. Pract., 157: 107843.
Samie, KA; Tabandeh, MR and Afrough, M (2020). Betaine ameliorates impaired steroidogenesis and apoptosis in mice granulosa cells induced by high glucose concentration. Systems Biol. Reprod. Med., 66: 400–409.
Sattar, AA; Matin, AA; Hadwan, MH; Hadwan, AM and Mohammed, RM (2024). Rapid and effective protocol to measure glutathione peroxidase activity. Bull. Nat. Res., Centre, 48: 100.
Shihab, EM and Kadhim, HM (2023). The impact of carvedilol on organ index, inflammatory mediators, oxidative stress parameters and skin markers in Dgalactose-induced aging mice. Int. J. Drug Delivery Technol., 13: 1017–1023.
Temidayo, SO and Du Plessis, SS (2018). Diabetes mellitus and male infertility. Asian Pacific J. Reprod., 7: 6–14.
Wang, P; Zhang, S; Lin, S and Lv, Z (2022). Melatonin ameliorates diabetic hyperglycaemia-induced impairment of Leydig cell steroidogenic function through activation of SIRT1 pathway. Reprod. Biol. Endocrinol., 20: 117.
Wikoff, WR; Anfora, AT; Liu, J; Schultz, PG; Lesley, SA; Peters, EC and Siuzdak, G (2009). Metabolomics analysis reveals large effects of gut microflora on mammalian blood metabolites. Proceed. Nat. Acad. Sci., 106: 3698–3703.
World Health Organization (2025). The cost of diabetes. Fact sheet No. 236 [Internet]. Geneva: WHO; 2002 [cited 2025 Sep 11]. Available from: https://www.who.int/mediacentre/factsheets/fs236/en/.
Wu, G; Lupton, JR; Turner, ND; Fang, YZ and Yang, S (2004). Glutathione metabolism and its implications for health. J. Nutr., 134: 489–492.
Zhang, B; Jiang, M; Zhao, J; Song, Y; Du, W and Shi, J (2022). The mechanism underlying the influence of indole-3-propionic acid: a relevance to metabolic disorders. Front. Endocrinol., 13: 841703.
Zhao, ZH; Xin, FZ; Xue, Y; Hu, Z; Han, Y; Ma, F; Zhou, D; Liu, XL; Cui, A and Liu, Z (2019). Indole-3-propionic acid inhibits gut dysbiosis and endotoxin leakage to attenuate steatohepatitis in rats. Exp. Mol. Med., 51: 1–14.
Zheng, Y; Ley, SH and Hu, FB (2018). Global aetiology and epidemiology of type 2 diabetes mellitus and its complications. Nature Rev. Endocrinol., 14: 88–98.
Caspian Journal of Veterinary Sciences
Volume 3, Issue 1
June 2026
Pages 33-43

  • Receive Date 13 September 2025
  • Revise Date 27 November 2025
  • Accept Date 06 January 2026