Postnatal Gelişme Dönemlerinde Capsaicin Uygulanan Fare Testislerinde TGF Beta-1’in İmmunohistokimyasal Lokalizasyonu

Transforme edici büyüme faktörü beta-1 (TGF Beta-1) testiküler fonksiyonların parakrin ya da otokrin düzenleyicilerinden biridir. Bu çalışmada, postnatal gelişme sürecinde Capsaicin (CAP) uygulanan farelerin testislerinde TGF Beta-1’in lokalizasyonu incelendi. Çalışmada 60 adet Swiss albino soyu erkek fare kullanıldı. Deney grubundaki 21 günlük farelere(n:30)75 gün boyunca her gün deri altı yolla 1mg/kg dozunda CAP enjekte edildi. Deney ve kontrol gruplarından 10’ar hayvan, 35. günü (puberte dönemi), 50. günü (puberte sonrası dönem) ve 75. günü (erişkin dönem) temsil edecek şekilde gruplara ayrıldı. Örneklenen testislerin tartımı sonrası rutin histolojik prosedür uygulandı. Çalışma süresince her gün kontrol ve deney grubu hayvanların canlı ağırlıkları tartıldı. Tüm deney gruplarındaki hayvanların canlı ağırlık ortalamalarının kontrol gruplarına oranla daha fazla olduğu ve bu iki grup arasında p<0,05 düzeyinde istatistiki önem saptandı. 35 ve 50 günlük deney gruplarının testis ağırlıklarının kontrol gruplarına oranla daha fazla olduğu gözlendi. Gözlenen farklılıklar arasında istatistiki bir öneme rastlanmadı.TGF Beta-1 immunreaksiyonu kontrol ve deney gruplarında, farklı boyanma yoğunluklarında, Leydig hücrelerinde ve genç spermatidlerde gözlenirken Sertoli hücrelerinde, spermatogonyumlarda ve erişkin spermatidlerde immunreaksiyon gözlenmedi. Genç spermatidlerdeki boyanma yoğunluğu her iki grupta da hemen hemen benzerdi, fakat kontrol gruplarında boyanma yoğunluğu yaş ile artarken deney gruplarındaki bu artış 75 günlük grupta gözlenmedi. Leydig hücrelerindeki boyanma yoğunluğu ise deney gruplarında yaş ile artış gösterirken kontrol gruplarında farklı boyanma yoğunlukları saptandı. Sonuç olarak CAP’ın, spermatogenez sırasında Leydig hücrelerindeki reseptörüne bağlanarak TGF Beta-1 immunreaksiyonunu arttırdığı sonucuna varıldı.

The Immunohistochemical Localization of TGF Beta-1 in Capsaicin Injected Mice Testis at the Postnatal Development Period

Transforming growth factor beta-1 (TGF Beta-1) is one of the paracrine or autocrine regulators of testicular function. In this study, we investigated the effect of Capsaicin (CAP) on the localization of TGF Beta-1 in the testis of mice treated with CAP during postnatal development. In the study, 60 Swiss albino male mice were used. The 21-day-old mice (n:30) in the experimental group were injected with CAP at 1 mg/kg subcutaneously daily for 75 days. Ten animals, each from the experimental and control groups, were divided into groups representing the 35th day (puberty), 50th day (post-puberty), and 75th day (adult). After weighing the testis, a routine histological procedure was performed. The body weights of the control and experimental group animals were measured daily during the study period. The mean body weights of all experimental groups were higher than the control groups, and we determined statistical significance at p<0.05 level between the groups. The mean testicular weights of the 35th and 50th experimental groups were higher than the 35th and 50th experimental groups, except for the 75th experimental group. No statistical significance was found between the groups. TGF Beta-1 immunoreaction was observed in Leydig cells and round spermatids at different intensities in all experimental and control groups. In contrast, no immunoreaction was observed in Sertoli cells, spermatogonia, and elongated spermatids. The staining intensity in round spermatids was almost similar in both groups. Still, the staining intensity increased with age in the control groups, while this increase in the 75th experimental groups was not observed. The staining intensity in Leydig cells increased with age in the experimental groups, while different staining intensities were found in the control groups. In conclusion, CAP increased TGF beta-1 immunoreaction by binding to its receptor in Leydig cells during spermatogenesis.

___

  • Adams EJ, Green JA, Clark AH, Youngson JH, 1999: Comparison of different scoring systems for immunohistochemical staining. J Clin Pathol, 52(1), 75–77.
  • Adaszek Ł, Gadomska D, Mazurek Ł, Łyp P, Madany J, Winiarczyk S, 2019: Properties of capsaicin and its utility in veterinary and human medicine. Res Vet Sci, 123, 14-19.
  • Dickson C, Webster DR, Johnson H, Cecilia Millena A, Khan SA, 2002: Transforming growth factor-beta effects on morphology of immature rat Leydig cells. Mol Cell Endocrinol, 195(1-2), 65-77.
  • Erdost H, Özer A, Yakışık M, Özfiliz N, Zık B, 2006: FSH and LH cells in the laying hens and cocks, fed with a diet containing red hot pepper. J Food Agr Env, 4(1), 119-123.
  • Erdost H, Ozfiliz N, Ozguden C, Gunes N, Onen S, Ilhan T, Ozer A, 2007: Expression of a capsaicin receptor (VR1) in the testes of mice after an application of capsaicin. Bull Vet Inst Pulawy, 51(4), 649-653.
  • Eurell AJ ve Frappier BL, 2006: Dellman’s Texbook of Veterinary Histology. 6th ed.,Blackwell Publishing, Iowa, USA.
  • Haagmans BL, Hoogerbrugge JW, Themmen AP, Teerds KJ, 2003: Rat testicular germ cells and Sertoli cells release different types of bioactive transforming growth factor beta in vitro. Reprod Biol Endocrinol, 5, 1-3.
  • Holzer P, 1991: Capsaicin as a tool for studying sensory neuron functions. Adv Exp Med Biol, 298(1), 3-16.
  • Ilhan T, Erdost H, 2013: Effects of capsaicin on testis ghrelin expression in mice. Biotech Histochem, 88(1), 8-10.
  • Itman C, Mendis S, Barakat B, Loveland KL, 2006: Focus on TGF-β signaling. All in the family: TGF-β family action in testis development. Reproduction, 132(2), 233-246.
  • Ingman WV, Robertson SA, 2002: Defining the actions of transforming growth factor beta in reproduction. Bioessays, 24(10), 904-914.
  • Jung JC, Park GT, Kim KH, Woo JH, An JM, Kim KC, Chung HY, Bae YS, Park JW, Kang SS, Lee YS, 2004: Differential expression of transforming growth factor-beta in the interstitial tissue of testis during aging. J Cell Biochem, 92(1),92-8.
  • Lui WY, Lee WM, Cheng CY, 2003: TGF-betas: their role in testicular function and Sertoli cell tight junction dynamics. Int J Androl, 26(3), 147-60.
  • Mizrak SC, Gadella BM, Erdost H, Ozer A, van Pelt AM, van Dissel-Emiliani FM, 2008: Spermatogonial stem cell sensitivity to capsaicin: an in vitro study. Reprod Biol Endocrinol, 6(52), 1-9.
  • Mullaney BP, Skinner MK, 1993: Transforming growth factor-beta (beta 1, beta 2, and beta 3) gene expression and action during pubertal development of the seminiferous tubule: potential role at the onset of spermatogenesis. Mol Endocrinol,7(1), 67-76.
  • Oh TW, Oh TW, Ohta F, 2003: Dose-dependent effect of capsaicin on endurance capacity in rats. Br J Nutr, 90(3), 515-20.
  • Özer A, Zık B, Erdost H, Özfiliz N, 2006: Histological investigations on the effects of feeding with a diet containing red hot pepper on the reproductive system organs of the cock. Turk J Vet Anim Sci, 30, 7-15.
  • Özgüden-Akkoç CG, İlhan T, Peker S, Zık B, 2017: Düşük doz kapsaisinin rat duodenumunda substans P ekspresyonu üzerine etkileri. Ankara Üniv Vet Fak Derg, 64(1), 31-36.
  • Özgüden-Akkoç CG, Asmaz ED, İlhan T, Zık B, 2018: Düşük doz kapsaicin uygulanan sıçanların ovaryumlarında TGF-Beta 1’in immunohistokimyasal yerleşimi. Erciyes Üniv Vet Fak Derg, 15(3), 238-246.
  • Park SG, Yon JM, Lin C, Gwon LW, Lee JG, Baek IJ, Lee BJ, Yun YW, Nam SY, 2017: Capsaicin attenuates spermatogenic cell death induced by scrotal hyperthermia through its antioxidative and anti-apoptotic activities. Andrologia, 49(5), 1-8.
  • Sharma SK, Vij AS, Sharma M, 2013: Mechanisms and clinical uses of capsaicin. Eur J Pharmacol, 720(1-3), 55-62.
  • Silva VEL, Cholewa JM, Jäger R, Zanchi NE, de Freitas MC, de Moura RC, Barros EML, Antunes BM, Caperuto EC, Ribeiro SLG, Lira FS, Pereira Dos Santos MA, Rossi FE, 2021: Chronic capsiate supplementation increases fat-free mass and upper body strength but not the inflammatory response to resistance exercise in young untrained men: a randomized, placebo-controlled and double-blind study. J Int Soc Sports Nutr, 18(1), 50.
  • Srinivasan K, 2016: Biological Activities of Red Pepper (Capsicum annuum) and Its Pungent Principle Capsaicin: A Review. Crit Rev Food Sci Nutr, 56(9), 1488-500.
  • Toukan N, Kulnik ST, Lewko A, ElShaer A, 2022: Therapeutic applications of capsaicin in humans to target conditions of the respiratory system: A scoping review. Respir Med, 194(2022), 106772.
  • Tütüncü Ş, Özfiliz N, 2011: Distribution of the vanilloid (capsaicin) receptor type 1 in the capsaicin treated rat ovaries on different sexual development periods. Rev Med Vet, 163(10), 460-7.
  • Watrin F, Scotto L, Assoian RK, Wolgemuth DJ,1991: Cell lineage specificity of expression of the murine transforming growth factor beta 3 and transforming growth factor beta 1 genes. Cell Growth Differ,2(1), 77-83.