Safranal ameliorates testicular ischemia-reperfusion injury in testicular torsion-detorsion rat model

Safranal mejora la lesión por isquemia-reperfusión testicular en un modelo de rata con detorsión de torsión testicular

Background: Testicular torsion-detorsion damage is a common ischemia-reperfusion injury brought on by an excess of reactive oxygen species. Reactive oxygen species may affect cellular differentiation by regulating gene expression. The heat shock protein 70-2 (HSP70-2) gene expression in the testis is essential for spermatogenesis. Safranal, the main bioactive ingredient isolated from Crocus sativus L., has potent antioxidant properties. Our current investigation examined the potential mechanism by which safranal could shield the testis from ischemia-reperfusion damage. Methods: Sixty Sprague-Dawley male rats were randomly assigned into the sham-operated control group, testicular ischemia-reperfusion group, and safranal-treated group. Testicular ischemia was achieved by twisting the left testis 720◦ counterclockwise and maintained for two hours. Reperfusion was created by counter-rotating the torsional left testis to its natural position. Rats in the safranal-treated group received an intraperitoneal injection of safranal at the onset of reperfusion. Testes were excised to examine the quantity of malondialdehyde (a sensitive indicator of reactive oxygen species), expression of the HSP70-2 protein, and the testicular spermatogenic activity. Results: Unilateral testicular ischemia-reperfusion significantly increased the levels of malondialdehyde in the ipsilateral testes compared to the control group. It also significantly reduced the expression of HSP70-2 protein and spermatogenic activity (p < 0.001). In addition, our investigation revealed that, in comparison to the testicular ischemia-reperfusion group, the ipsilateral testes of the safranal-treated group had significantly lower levels of malondialdehyde and had significantly higher HSP70-2 expression and levels of spermatogenic function (p < 0.01). Conclusions: These results suggest that via lowering reactive oxygen species levels and increasing HSP70-2 expression, safranal protects against testicular torsion/detorsion-induced ischemia/reperfusion injury.

Resumen

Antecedentes: La lesión después de la detorsión de la torsión testicular es típica de isquemia-reperfusión causada por la producción excesiva de especies reactivas de oxígeno. Las especies reactivas de oxígeno pueden afectar la diferenciación celular regulando la expresión génica. La expresión del gen de la proteína de choque térmico 70-2 (HSP70-2) en los testículos es crucial para la espermatogénesis. El azafrán es el principal activo biológico extraído del Crocus sativus L. y tiene fuertes propiedades antioxidantes. En este estudio, probamos si safranal protege los testículos de lesiones por isquemia-reperfusión, así como posibles mecanismos. Métodos: Sesenta ratas macho Sprague-Dawley fueron divididas aleatoriamente en tres grupos: grupo de control de cirugía falsa, grupo de isquemia-reperfusión testicular y grupo de tratamiento con safranal. La isquemia testicular se logra girando 720◦ del testículo izquierdo en sentido contrario a las agujas del reloj y manteniéndolo durante dos horas. La reinyección se produce girando el testículo izquierdo invertido a su posición natural. En el grupo de tratamiento con safranal, las ratas se inyectaron safranal en la cavidad abdominal al comienzo de la reinyección. Se extirparon los testículos para analizar la concentración de malondialdehído (biomarcadores sensibles al oxígeno activo), la expresión de la proteína HSP70-2 y la función espermatogénica testicular. Resultados: En comparación con el grupo control del grupo, la espasmos-reinyección estimulados por los testículos unilaterales tuvo un efecto significativo en la concentración de malondialdehído, la expresión de la proteína HSP70-2 y la función espermática en sujetos del mismo lado (p < 0.001). Además, la expresión de HSP70-2 y la concentración de malondialdehído en la prueba ipsilateral se estudiaron significativamente en el grupo de tratamiento de reinyección testicular en comparación con el grupo de reinyección testicular (p < 0.01). Conclusiones: Estos resultados sugieren que al reducir los niveles de especies reactivas de oxígeno y aumentar la expresión de HSP70-2, el guayanal protege los testículos de lesiones por isquemia/reperfusión inducidas por torsión/detorsión.

Safranal; Testicular ischemia-reperfusion injury; Testicular torsion-detorsion; Malondi-aldehyde; Heat shock protein 70-2; Testicular spermatogenic functio

Palabras Clave

Safranal; Lesión por isquemia-reperfusión testicular; Torsión testicular; Malondialdehído; Proteína de choque térmico 70-2; Función espermatogénica testicular

[1] Abu-Baih RH, Abu-Baih DH, Abdel-Hafez SMN, Fathy M. Activation of SIRT1/Nrf2/HO-1 and Beclin-1/AMPK/mTOR autophagy pathways by eprosartan ameliorates testicular dysfunction induced by testicular torsion in rats. Scientific Reports. 2024; 14: 12566.

[2] Cigsar Kuzu EB, Oztan MO, Guney N, Koyluoglu G. Reassessment of a decade of experience on testicular torsion: are we making a mistake? Urologia Internationalis. 2022; 106: 1100–1106.

[3] Zvizdic Z, Aganovic A, Milisic E, Jonuzi A, Zvizdic D, Vranic S. Duration of symptoms is the only predictor of testicular salvage following testicular torsion in children: a case-control study. American Journal of Emergency Medicine. 2021; 41: 197–200.

[4] He KX, Ning JZ, Li W, Cheng F. Emodin alleviates testicular ischemia-reperfusion injury through the inhibition of NLRP3-mediated pyroptosis. Tissue & Cell. 2023; 82: 102069.

[5] Demir EA, Demir S, Kazaz IO, Kucuk H, Alemdar NT, Buyuk A, et al. Arbutin abrogates testicular ischemia/reperfusion injury in rats through repression of inflammation and ER stress. Tissue & Cell. 2023; 82: 102056.

[6] Ayobami Afolabi O, Adebola Alabi B, Adedamola Ajike R, Simeon Oyekunle O, Adegoke W, Adebayo Ojetola A. Evaluation of testicular torsion management in Ogbomoso, South-Western Nigeria and surgical detorsion-augmented treatment with phytochemical fractions of Corchorus olitorius leaf in experimental rats. Saudi Journal of Biological Sciences. 2023; 30: 103495.

[7] Demir S, Kazaz IO, Kerimoglu G, Alemdar NT, Colak F, Arici T, et al. Gallic acid attenuates torsion/detorsion-induced testicular injury in rats through suppressing of HMGB1/NF-κB axis and endoplasmic reticulum stress. Revista Internacional de Andrologia. 2024; 22: 1–7.

[8] Bej E, Volpe AR, Cesare P, Cimini A, d’Angelo M, Castelli V. Therapeutic potential of saffron in brain disorders: from bench to bedside. Phytotherapy Research. 2024; 38: 2482–2495.

[9] Yang W, Wei Y, Sun J, Yao C, Ai F, Ding H. Safranal exerts a neuroprotective effect on Parkinson’s disease with suppression of NLRP3 inflammation activation. Molecular Biology Reports. 2024; 51: 593.

[10] Abdalla Y, Abdalla A, Hamza AA, Amin A. Safranal prevents liver cancer through inhibiting oxidative stress and alleviating inflammation. Frontiers in Pharmacology. 2022; 12: 777500.

[11] Sadeghnia HR, Shaterzadeh H, Forouzanfar F, Hosseinzadeh H. Neuroprotective effect of safranal, an active ingredient of Crocus sativus, in a rat model of transient cerebral ischemia. Folia Neuropathologica. 2017; 55: 206–213.

[12] Ozkececi ZT, Gonul Y, Yuksel Y, Karavelioglu A, Tunay K, Gulsari Y, et al. Investigation of the effect of safranal and crocin pre-treatment on hepatic injury induced by infrarenal aortic occlusion. Biomedicine & Pharmacotherapy. 2016; 83: 160–166.

[13] Bharti S, Golechha M, Kumari S, Siddiqui KM, Arya DS. Akt/GSK-3β/eNOS phosphorylation arbitrates safranal-induced myocardial protection against ischemia-reperfusion injury in rats. European Journal of Nutrition. 2012; 51: 719–727.

[14] Hosseinzadeh H, Modaghegh MH, Saffari Z. Crocus sativus L. (Saffron) extract and its active constituents (crocin and safranal) on ischemia-reperfusion in rat skeletal muscle. Evidence-based Complementary and Alternative Medicine. 2009; 6: 343–350.

[15] Wei SM, Huang YM. Attenuation effect of salvianolic acid B on testicular ischemia-reperfusion injury in rats. Oxidative Medicine and Cellular Longevity. 2022; 2022: 7680182.

[16] Ohkawa H, Ohishi N, Yagi K. Assay for lipid peroxides in animal tissues by thiobarbituric acid reaction. Analytical Biochemistry. 1979; 95: 351–358.

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

[18] Wei SM, Huang YM, Qin ZQ. Salidroside exerts beneficial effect on testicular ischemia-reperfusion injury in rats. Oxidative Medicine and Cellular Longevity. 2022; 2022: 8069152.

[19] 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.

[20] Sharp VJ, Kieran K, Arlen AM. Testicular torsion: diagnosis, evaluation, and management. American Family Physician. 2013; 88: 835–840.

[21] Sessions AE, Rabinowitz R, Hulbert WC, Goldstein MM, Mevorach RA. Testicular torsion: direction, degree, duration and disinformation. Journal of Urology. 2003; 169: 663–665.

[22] Xu LZ, He KX, Ning JZ, Cheng F. Oleuropein attenuates testicular ischemia-reperfusion by inhibiting apoptosis and inflammation. Tissue & Cell. 2022; 78: 101876.

[23] Dix DJ, Allen JW, Collins BW, Poorman-Allen P, Mori C, Blizard DR, et al. HSP70-2 is required for desynapsis of synaptonemal complexes during meiotic prophase in juvenile and adult mouse spermatocytes. Development. 1997; 124: 4595–4603.

[24] Feng HL, Sandlow JI, Sparks AE. Decreased expression of the heat shock protein hsp70-2 is associated with the pathogenesis of male infertility. Fertility and Sterility. 2001; 76: 1136–1139.

[25] Dix DJ, Allen JW, Collins BW, Mori C, Nakamura N, Poorman-Allen P, et al. Targeted gene disruption of Hsp70-2 results in failed meiosis, germ cell apoptosis, and male infertility. Proceedings of the National Academy of Sciences of the United States of America. 1996; 93: 3264–3268.

[26] Dix DJ. Hsp70 expression and function during gametogenesis. Cell Stress & Chaperones. 1997; 2: 73–77.

[27] Dix DJ, Hong RL. Protective mechanisms in germ cells: stress proteins in spermatogenesis. Advances in Experimental Medicine and Biology. 1998; 444: 137–144.

[28] Eddy EM. Role of heat shock protein HSP70-2 in spermatogenesis. Reviews of Reproduction. 1999; 4: 23–30.

[29] Tien Kuo M, Savaraj N. Roles of reactive oxygen species in hepatocarcinogenesis and drug resistance gene expression in liver cancers. Molecular Carcinogenesis. 2006; 45: 701–709.

[30] Assimopoulou AN, Sinakos Z, Papageorgiou VP. Radical scavenging activity of Crocus sativus L. extract and its bioactive constituents. Phytotherapy Research. 2005; 19: 997–1000.

[31] Pouchieu C, Pourtau L, Brossaud J, Gaudout D, Corcuff JB, Capuron L, et al. Acute effect of a saffron extract (Safr’InsideTM) and its main volatile compound on the stress response in healthy young men: a randomized, double blind, placebo-controlled, crossover Study. Nutrients. 2023; 15: 2921.

[32] Pachikian BD, Copine S, Suchareau M, Deldicque L. Effects of saffron extract on sleep quality: a randomized double-blind controlled clinical trial. Nutrients. 2021; 13: 1473.

[33] Kell G, Rao A, Beccaria G, Clayton P, Inarejos-García AM, Prodanov M. affron® a novel saffron extract (Crocus sativus L.) improves mood in healthy adults over 4 weeks in a double-blind, parallel, randomized, placebo-controlled clinical trial. Complementary Therapies in Medicine. 2017; 33: 58–64.

[34] Bahadir GB, Gollu G, Ilkay H, Bagriacik U, Hasirci N, Bingol-Kologlu M. LOCAL-IGF-1 and GH application improves germ cell histology, spermatogenesis and fertility after experimental testicular torsion and detorsion. Journal of Pediatric Urology. 2022; 18: 410.e1–410.e8.

[35] Antonuccio P, Pallio G, Marini HR, Irrera N, Romeo C, Puzzolo D, et al. Involvement of hypoxia-inducible factor 1-α in experimental testicular ischemia and reperfusion: effects of polydeoxyribonucleotide and selenium. International Journal of Molecular Sciences. 2022; 23: 13144.

[36] Almarzouq D, Al-Maghrebi M. NADPH oxidase-mediated testicular oxidative imbalance regulates the TXNIP/NLRP3 inflammasome axis activation after ischemia reperfusion injury. Antioxidants. 2023; 12: 145.

[37] Jafarova Demirkapu M, Karabag S, Akgul HM, Mordeniz C, Yananli HR. The effects of etomidate on testicular ischemia reperfusion injury in ipsilateral and contralateral testes of rats. European Review for Medical and Pharmacological Sciences. 2022; 26: 211–217.

[38] Akhigbe RE, Odetayo AF, Akhigbe TM, Hamed MA, Ashonibare PJ. Pathophysiology and management of testicular ischemia/reperfusion injury: lessons from animal models. Heliyon. 2024; 10: e27760.

[39] Yilmaz N, Yildiz A, Tanbek K, Kisaoglu A, Yilmaz U, Kose E. Protective effect of astaxanthin on testis torsion/detorsion injury through modulation of autophagy. Revista Internacional de Andrologia. 2024; 22: 29–37.

[40] Azizoğlu M, Arslan S, Gökalp-Özkorkmaz E, Aşır F, Basuguy E, Okur MH, et al. Protective effects of Passiflora Incarnata on ischemia-reperfusion injury in testicular torsion: an experimental study in a rat model. Cirugia y Cirujanos. 2024; 92: 165–173.

[41] Yilmaz N, Hudaykulıyeva J, Gul S. Phoenixin-14 may ameliorate testicular damage caused by torsion-detorsion by reducing oxidative stress and inflammation in prepubertal rats. Tissue & Cell. 2024; 88:102405.

[42] Ebrahimi M, Abtahi-Eivary SH, Brazvan B, Shokoohi M, Soltani M, Rostamian M, et al. Safranal ameliorates ischemic/reperfusion injury induced by testicular torsion in rat. Physiology and Pharmacology. 2023; 27: 403–416.