Regulation of STAT3 and NF-κB signaling pathways by trans-Anethole in testicular ischemia-reperfusion injury and its gonadoprotective effect

Regulación de las vías de señalización de STAT3 y NF-κB por trans-Anetol en la lesión por isquemia-reperfusión testicular y su efecto gonadoprotector

Testicular ischemia reperfusion (I/R) injury is a significant urological problem where clinical interventions may be inadequate, and the antioxidants might be potential co-treatment modalities. This study examined the gonadoprotective effect of trans-Anethole in testicular I/R injury. Twenty-eight male rats were divided into four groups. Rats in the I/R, I/R + t100, I/R + t200 groups underwent bilateral testicular I/R injury. The I/R + t100 and I/R + t200 groups received 100 or 200 mg/kg trans-Anethole at the 2nd hour of ischemia. Microscopic evaluations demonstrated that testicular I/R injury leads to severe testicular degeneration. Tissue oxidative stress, pro-apoptotic Bcl-2 associated X (Bax) and Caspase 3, pro-inflammatory Tumor necrosis factor-alpha (TNF-α), Interleukin-1 beta (IL-1β) and Interleukin 6 (IL-6) cytokines levels were significantly (p < 0.05) upregulated when compared to the Control group. Additionally, transcription factors Signal transducer and activator of transcription 3 (STAT3) and Nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB) levels increased significantly (p < 0.05) compared to the Control group. Tissue disrupted parameters in the I/R + t200 group were significantly different (p < 0.05) from the I/R group, contrasting with the slight improvement in the I/R + t100 group. The STAT3 and NF-κB expression levels in the I/R + t200 group were significantly suppressed (p < 0.05) compared to the I/R group. In conclusion, our study indicates that trans-Anethole could enhance gonadoprotective activity in testicular I/R injury, potentially involving transcription factors STAT3 and NF-κB. However, before the consumption of trans-Anethole-containing natural or manufactured goods, the potential benefits and side effects should be carefully evaluated.

Resumen

La lesión por isquemia-reperfusión (I/R) testicular es un problema urológico importante en el que las intervenciones clínicas pueden resultar inadecuadas y los antioxidantes podrían ser posibles modalidades de cotratamiento. Este estudio examinó el efecto gonadoprotector del trans-Anetol en la lesión por I/R testicular. Se dividieron veintiocho ratas macho en cuatro grupos. Las ratas en los grupos I/R, I/R + t100, I/R + t200 sufrieron una lesión por I/R testicular bilateral. Los grupos I/R + t100 e I/R + t200 recibieron 100 o 200 mg/kg de trans-Anetol en la segunda hora de isquemia. Las evaluaciones microscópicas demostraron que la lesión por I/R testicular conduce a una degeneración testicular grave. El estrés oxidativo tisular, Bcl-2 associated X (Bax) y Caspase 3 proapoptóticas, las citocinas proinflamatorias Tumor necrosis factor-alpha (TNF-α), Interleukin-1 beta (IL-1β) e Interleukin 6 (IL-6) aumentaron significativamente (p < 0.05) en comparación con el grupo de control. Además, los niveles de los factores de transcripción Signal transducer and activator of transcription 3 (STAT3) y Nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB) aumentaron significativamente (p < 0.05) en comparación con el grupo de control. Los parámetros de tejido alterado en el grupo I/R + t200 fueron significativamente diferentes (p < 0.05) de los del grupo I/R, en contraste con la ligera mejoría en el grupo I/R + t100. Los niveles de expresión de STAT3 y NF-κB en el grupo I/R + t200 se suprimieron significativamente (p < 0.05) en comparación con el grupo I/R. En conclusión, nuestro estudio indica que el trans-Anetol podría mejorar la actividad gonadoprotectora en la lesión por I/R testicular, involucrando potencialmente a los factores de transcripción STAT3 y NF-κB. Sin embargo, antes de consumir productos naturales o manufacturados que contengan trans-Anetol, se deben evaluar cuidadosamente los posibles beneficios y efectos secundarios.

Testis; Ischemia reperfusion injury; trans-Anethole; Apoptosis; Inflammation

Palabras Clave

Testículo; Lesión por isquemia-reperfusión; trans-Anetol; Apoptosis; Inflamación

[1] Laher A, Ragavan S, Mehta P, Adam A. Testicular torsion in the emergency room: a review of detection and management strategies. Open Access Emergency Medicine. 2020; 12: 237–246.

[2] Raisi A, Davoodi F. Testicular torsions in veterinary medicine. Veterinary Research Communications. 2022; 46: 301–313.

[3] Greear GM, Romano MF, Katz MH, Munarriz R, Rague JT. Testicular torsion: epidemiological risk factors for orchiectomy in pediatric and adult patients. International Journal of Impotence Research. 2021; 33: 184–190.

[4] Lacy A, Smith A, Koyfman A, Long B. High risk and low prevalence diseases: testicular torsion. The American Journal of Emergency Medicine. 2023; 66: 98–104.

[5] Abdelzaher WY, Mostafa-Hedeab G, Sayed AboBakr Ali AH, Fawzy MA, Ahmed AF, Bahaa El-deen MA, et al. Idebenone regulates sirt1/Nrf2/TNF-α pathway with inhibition of oxidative stress, inflammation, and apoptosis in testicular torsion/detorsion in juvenile rats. Human & Experimental Toxicology. 2022; 41: 09603271221102515.

[6] Wei S, Huang Y. Baicalein alleviates testicular ischemia-reperfusion injury in a rat model of testicular torsion-detorsion. Oxidative Medicine and Cellular Longevity. 2022; 2022: 1603469.

[7] Seker U, Kavak DE, Guzel BC, Baygeldi SB, Yuksel M, Unay Demirel O, et al. Targeting soluble guanylate cyclase with Riociguat has potency to alleviate testicular ischaemia reperfusion injury via regulating various cellular pathways. Andrologia. 2022; 54: e14616.

[8] Onishi A, Chiba K, Kawamura S, Sato K, Kaku Y, Okada K, et al. Dimethyl fumarate protects rats against testicular ischemia–reperfusion injury. Andrologia. 2023; 2023: 7086044.

[9] Yuksel S, Erginel B, Bingul I, Ozluk Y, Karatay H, Aydın F, et al. The effect of hydrogen sulfide on ischemi̇a/reperfusion injury in an experimental testicular torsion model. Journal of Pediatric Urology. 2022; 18: 16.e1–16.e7.

[10] Mohamed MZ, Morsy MA, Mohamed HH, Hafez HM. Paeonol protects against testicular ischaemia‐reperfusion injury in rats through inhibition of oxidative stress and inflammation. Andrologia. 2020; 52: e13599.

[11] Ghasemi-Dehnoo M, Safari AA, Rahimi-Madiseh M, Lorigooini Z, Moradi MT, Amini-Khoei H. Anethole ameliorates acetic acid-induced colitis in mice: anti-inflammatory and antioxidant effects. Evidence-Based Complementary and Alternative Medicine. 2022; 2022: 9057451.

[12] Sharafan M, Jafernik K, Ekiert H, Kubica P, Kocjan R, Blicharska E, et al. Illicium verum (star anise) and trans-Anethole as valuable raw materials for medicinal and cosmetic applications. Molecules. 2022; 27: 650.

[13] Younis NS, Mohamed ME. Anethole pretreatment modulates cerebral ischemia/reperfusion: the role of JNK, p38, MMP-2 and MMP-9 pathways. Pharmaceuticals. 2023; 16: 442.

[14] Mohamed ME, Kandeel M, Abd El-Lateef HM, El-Beltagi HS, Younis NS. The protective effect of anethole against renal ischemia/reperfusion: the role of the TLR2,4/MYD88/NFκB pathway. Antioxidants. 2022; 11: 535.

[15] Ileriturk M, Kandemir O, Akaras N, Simsek H, Genc A, Kandemir FM. Hesperidin has a protective effect on paclitaxel-induced testicular toxicity through regulating oxidative stress, apoptosis, inflammation and endoplasmic reticulum stress. Reproductive Toxicology. 2023; 118: 108369.

[16] He B, Chen D, Zhang X, Yang R, Yang Y, Chen P, et al. Oxidative stress and ginsenosides: an update on the molecular mechanisms. Oxidative Medicine and Cellular Longevity. 2022; 2022: 9299574.

[17] Qin C, Yang S, Chu Y-H, Zhang H, Pang X-W, Chen L, et al. Signaling pathways involved in ischemic stroke: molecular mechanisms and therapeutic interventions. Signal Transduction and Targeted Therapy. 2022; 7: 215.

[18] Alnajem A, Al-Maghrebi M. The regulatory effects of JAK2/STAT3 on spermatogenesis and the redox keap1/nrf2 Axis in an animal model of testicular ischemia reperfusion injury. Cells. 2023; 12: 2292.

[19] Xu Y, Zhang J, Gao F, Cheng W, Zhang Y, Wei C, et al. Engeletin alleviates cerebral ischemia reperfusion‐induced neuroinflammation via the HMGB1/TLR4/NF‐κB network. Journal of Cellular and Molecular Medicine. 2023; 27: 1653–1663.

[20] Keskin E, Erdogan A, Suleyman H, Yazici GN, Sunar M, Gul MA. Effect of sunitinib on testicular oxidative and proinflammatory damage induced by ischemia–reperfusion in rats. Revista Internacional de Andrología. 2022; 20: S17–S23.

[21] Fagundes VH, Pinho RJ, Wiirzler LA, Kimura E, Bersani-Amado CA, Cuman RK. High performance liquid chromatography method for the determination of anethole in rat plasma. Tropical Journal of Pharmaceutical Research. 2014; 13: 793–799.

[22] Sağır S, Şeker U, Pekince Özener M, Yüksel M, Demir M. Oxidative stress, apoptosis, inflammation, and proliferation modulator function of visnagin provide gonadoprotective activity in testicular ischemia-reperfusion injury. European Review for Medical and Pharmacological Sciences. 2023; 27: 9968–9977.

[23] Seker U, Guzel BC, Sener D, Baygeldi SB, Yuksel M, Demirel OU, et al. Nephroprotective effect of aloe Vera extract with regulation of oxidative stress, apoptosis and aquaporin 3 expression levels in streptozotocin induced diabetic rats. Journal of Faculty of Pharmacy of Ankara University. 2023; 47: 438–449.

[24] Seker U, Nergiz Y, Aktas A, Akkus M, Ozmen MF, Uyar E, et al. Trolox is more successful than allopurinol to reduce degenerative effects of testicular ischemia/reperfusion injury in rats. Journal of Pediatric Urology. 2020; 16: 465.e461–465.e8.

[25] Ito Y, Unagami M, Yamabe F, Mitsui Y, Nakajima K, Nagao K, et al. A method for utilizing automated machine learning for histopathological classification of testis based on Johnsen scores. Scientific Reports. 2021; 11: 9962.

[26] Johnsen SG. Testicular biopsy score count—a method for registration of spermatogenesis in human testes: normal values and results in 335 hypogonadal males. Hormones. 1970; 1: 2–25.

[27] Hirata K, Ohira S, Tone S, Nakatsuka T, Morinaka H, Takasaki H, et al. Pathological analysis of spermatic dysfunction in testicular ischemia-reperfusion injury. The Journal of Sexual Medicine. 2022; 19: S207.

[28] Arena S, Iacona R, Antonuccio P, Russo T, Salvo V, Gitto E, et al. Medical perspective in testicular ischemia‑reperfusion injury. Experimental and Therapeutic Medicine. 2017; 13: 2115–2122.

[29] Erdem AO, Ozkisacik S, Mersinli B, Şirinyildiz F, Ek R, Culhaci N, et al. Long-term protective effects of the combination of intermittent reperfusion and hypothermia on reperfusion injury in an experimental testicular torsion model. Journal of Pediatric Surgery. 2021; 56: 2037–2044.

[30] Karaguzel E, Kadihasanoglu M, Kutlu O. Mechanisms of testicular torsion and potential protective agents. Nature Reviews Urology. 2014; 11: 391–399.

[31] Cho H-I, Kim K-M, Kwak JH, Lee SK, Lee S-M. Protective mechanism of anethole on hepatic ischemia/reperfusion injury in mice. Journal of Natural Products. 2013; 76: 1717–1723.

[32] Lu J, Hou W, Yang S, Chen D, Wang F, Liu L, et al. Trans-anethole pretreatment ameliorates hepatic ischemia–reperfusion injury via regulation of soluble epoxide hydrolase. International Immunopharmacology. 2023; 124: 110809.

[33] Ryu S, Seol GH, Park H, Choi I-Y. Trans-anethole protects cortical neuronal cells against oxygen–glucose deprivation/reoxygenation. Neurological Sciences. 2014; 35: 1541–1547.

[34] Vastegani SM, Khoshnam SE, Mansouri E, Hajipour S, Ghafouri S, Bakhtiari N, et al. Neuroprotective effect of anethole against rotenone induced non-motor deficits and oxidative stress in rat model of Parkinson’s disease. Behavioural Brain Research. 2023; 437: 114100.

[35] Tehranipour M, Gheysari AH, Dahrazma R, Hoseini S, Rajabniya H, Amghani MK. The angiogenic-antioxidant activity and anti-diabetic histological impacts of chitosan-coated anethole Nano-phytosomes (CCA-NPs) on the diabetic rats’ pancreatic tissue and blood biochemistry indices. BioNanoScience. 2024; 14: 1473–1481.

[36] Matboli M, Hasanin AH, Hamady S, Khairy E, Mohamed RH, Aboul-Ela YM, et al. Anti-inflammatory effect of trans-anethol in a rat model of myocardial ischemia-reperfusion injury. Biomedicine & Pharmacotherapy. 2022; 150: 113070.

[37] Younis NS, Mohamed ME. Anethole’s effects against myocardial infarction: the role of TLR4/NFκB and Nrf2/HO1 pathways. Chemico-Biological Interactions. 2022; 360: 109947.

[38] Yildizhan E, Akkus M, Ulger B, Asır F, Söker S, Gündüz E, et al. Protective effectiveness of anise against testicular ischemia and reperfusion injury: an experimental study in rats. Polish Journal of Veterinary Sciences. 2023; 26: 249–255.

[39] Chainy GB, Manna SK, Chaturvedi MM, Aggarwal BB. Anethole blocks both early and late cellular responses transduced by tumor necrosis factor: effect on NF-κB, AP-1, JNK, MAPKK and apoptosis. Oncogene. 2000; 19: 2943–2950.

[40] Sung Y-Y, Kim YS, Kim HK. Illicium verum extract inhibits TNF-α- and IFN-γ-induced expression of chemokines and cytokines in human keratinocytes. Journal of Ethnopharmacology. 2012; 144: 182–189.