Journal of Animal Reproduction and Biotechnology (한국동물생명공학회지)

The Korean Society of Animal Reproduction and Biotechnology (한국동물생명공학회)
권호 표지
DOI QR코드

DOI QR Code

Regulation of the plasminogen activator activity and inflammatory environment via transforming growth factor-beta regulation of sperm in porcine uterine epithelial cells

  • Kim, Su-jin (College of Animal Life Sciences, Kangwon National University) ;
  • Cheong, Hee-Tae (College of Veterinary Medicine, Kangwon National University) ;
  • Park, Choon-keun (College of Animal Life Sciences, Kangwon National University)
  • Received : 2020.11.13
  • Accepted : 2020.12.05
  • Published : 2020.12.31
Download PDF
⟨ Previous Next ⟩

Abstract

The aims of the present study were to confirm that regulation of the PA and environment via TGF-β regulation of sperm by Percoll-separated in porcine uterine epithelial cells. And, it was performed to identify the cytokines (TGF-β1, 2 and 3, TGF-β receptor1 and 2; interleukin, IL-6, IL-8) and PA-related genes (urokinase-PA, uPA; tissue-PA, tPA; PA inhibitor, PAI; uPA-receptor, uPAR) by spermatozoa. The experiment used porcine uterus epithelial cells (pUECs) and uterine tissue epithelial cells, Boar sperm were separated by discontinuous Percoll density gradient (45/90%), and tissues were co-incubated with spermatozoa, followed by real-time PCR. PA activity was measured of sperm by discontinuous Percoll density gradient (45/90%) for 24 hours. To measure viability and acrosome damage of sperm double stained propidium iodide (PI) and SYBR-14 or FITC-PNA were used. In results, binding ratio of Percoll-separated sperm was found no differences, but sperms isolated from 90% Percoll layer reduced PA activity (p < 0.05). when co-cultured sperm selected Percoll in porcine uterus tissues epithelial cells, 90% layer sperm increased TGF-β R1, contrastively tPA and PAI-1 in comparison with control (p < 0.05). 45% sperm was decreased the expression of uPA (p < 0.05). TGF-β decreased PA activity in the supernatant collected from pUECs (p < 0.05). Especially, The group including uPA, PAI-1 were induce sperm intact, while it was reduced in sperm damage when compared to control (p < 0.05). Also, there was no significant difference group of tPA and tPA+I in the dead sperm and acrosome damage compared to control. The expression of tPA and PAI showed a common response. Percoll-separated spermatozoa in 90% layer reduced tPA and IL-related gene mRNA expression. Thus, Percoll-sparated sperm in 90% layer show that it can suppress inflammation through increased expression of TGF-β and downregulation of PA and IL in epithelial cells compared to 45% layer Percoll.

Keywords

References

  1. Alsina RB and Mukhametova LI. 2014. [Structure and functions of plasminogen/plasmin system]. Bioorg. Khim. 40:642-657. Russian.
  2. Ardon F, Markello RD, Hu L, Deutsch ZI, Tung CK, Wu M, Suarez SS. 2016. Dynamics of bovine sperm interaction with epithelium differ between oviductal isthmus and ampulla. Biol. Reprod. 95:90. https://doi.org/10.1095/biolreprod.116.140632
  3. Barranco I, Padilla L, Perez-Patino C, Vazquez JM, Martinez EA, Rodriguez-Martinez H, Roca J, Parrilla I. 2019. Seminal plasma cytokines are predictive of the outcome of boar sperm preservation. Front. Vet. Sci. 6:436. https://doi.org/10.3389/fvets.2019.00436
  4. Castellino FJ and Ploplis VA. 2005. Structure and function of the plasminogen/plasmin system. Thromb. Haemost. 93:647-654. https://doi.org/10.1267/THRO05040647
  5. Castelo Branco MA, Castelo Branco YNTC, Moraes Junior FJ, Barros FN, Barcante FPS, Carvalho GMC, Melo Evangelista LS, Abreu-Silva AL, Sousa Filho MA, Souza JAT. 2017. Plasminogen activator inhibitor 1 and Antipain preserve acrosome integrity of bovine spermatozoa during cryopreservation. Arq. Bras. Med. Vet. Zootec. 69:1114-1124. https://doi.org/10.1590/1678-4162-9252
  6. Ebisch IM, Thomas CM, Wetzels AM, Willemsen WN, Sweep FC, Steegers-Theunissen RP. 2008. Review of the role of the plasminogen activator system and vascular endothelial growth factor in subfertility. Fertil. Steril. 90:2340-2350. https://doi.org/10.1016/j.fertnstert.2007.10.026
  7. Espana F, Gilabert J, Estelles A, Romeu A, Aznar J, Cabo A. 1991. Functionally active protein C inhibitor/plasminogen activator inhibitor-3 (PCI/PAI-3) is secreted in seminal vesicles, occurs at high concentrations in human seminal plasma and complexes with prostate-specific antigen. Thromb. Res. 64:309-320. https://doi.org/10.1016/0049-3848(91)90002-E
  8. Fatehi AN, Bevers MM, Schoevers E, Roelen BA, Colenbrander B, Gadella BM. 2006. DNA damage in bovine sperm does not block fertilization and early embryonic development but induces apoptosis after the first cleavages. J. Androl. 27:176-188. https://doi.org/10.2164/jandrol.04152
  9. Grullon LA, Gadea J, Mondejar I, Matas C, Romar R, Coy P. 2013. How is plasminogen/plasmin system contributing to regulate sperm entry into the oocyte? Reprod. Sci. 20:1075-1082. https://doi.org/10.1177/1933719112473657
  10. Kotilainen T, Huhtinen M, Katila T. 1994. Sperm-induced leukocytosis in the equine uterus. Theriogenology 41:629-636. https://doi.org/10.1016/0093-691X(94)90173-G
  11. Li Q. 2014. Transforming growth factor β signaling in uterine development and function. J. Anim. Sci. Biotechnol. 5:52. https://doi.org/10.1186/2049-1891-5-52
  12. Lopez-Ubeda R, Garcia-Vazquez FA, Gadea J, Matas C. 2017. Oviductal epithelial cells selected boar sperm according to their functional characteristics. Asian J. Androl. 19:396-403. https://doi.org/10.4103/1008-682X.173936
  13. Marey MA, Matsukawa H, Sasaki M, Ezz MA, Yousef MS, Takahashi KI, Miyamoto A. 2020. Bovine oviduct epithelial cells suppress the phagocytic activity of neutrophils towards sperm but not for bacteria in vitro: immunofluorescence and electron microscopic observations. Histol. Histopathol. 35:589-597.
  14. Matas C, Coy P, Romar R, Marco M, Gadea J, Ruiz S. 2003. Effect of sperm preparation method on in vitro fertilization in pigs. Reproduction 125:133-141. https://doi.org/10.1530/reprod/125.1.133
  15. Parrish JJ, Krogenaes A, Susko-Parrish JL. 1995. Effect of bovine sperm separation by either swim-up or Percoll method on success of in vitro fertilization and early embryonic development. Theriogenology 44:859-869. https://doi.org/10.1016/0093-691X(95)00271-9
  16. Rotello RJ, Lieberman RC, Purchio AF, Gerschenson LE. 1991. Coordinated regulation of apoptosis and cell proliferation by transforming growth factor beta 1 in cultured uterine epithelial cells. Proc. Natl. Acad. Sci. U. S. A. 88:3412-3415. https://doi.org/10.1073/pnas.88.8.3412
  17. Ryan TJ, Lai L, Malik AB. 1992. Plasmin generation induces neutrophil aggregation: dependence on the catalytic and lysine binding sites. J. Cell. Physiol. 151:255-261. https://doi.org/10.1002/jcp.1041510206
  18. Sa SJ, Rhee HH, Cheong HT, Yang BK, Park CK. 2006. Effects of plasmin on sperm-oocyte interactions during in vitro fertilization in the pig. Anim. Reprod. Sci. 95:273-282. https://doi.org/10.1016/j.anireprosci.2006.01.017
  19. Sharkey DJ, Macpherson AM, Tremellen KP, Mottershead DG, Gilchrist RB, Robertson SA. 2012. TGF-β mediates proinflammatory seminal fluid signaling in human cervical epithelial cells. J. Immunol. 189:1024-1035. https://doi.org/10.4049/jimmunol.1200005
  20. Smokovitis A, Kokolis N, Alexopoulos C, Alexaki E, Eleftheriou E. 1987. Plasminogen activator activity, plasminogen activator inhibition and plasmin inhibition in spermatozoa and seminal plasma of man and various animal species-effect of plasmin on sperm motility. Fibrinolysis 1:253-257. https://doi.org/10.1016/0268-9499(87)90045-2
  21. Waberski D, Magnus F, Ardon F, Petrunkina AM, Weitze KF, Topfer-Petersen E. 2006. Binding of boar spermatozoa to oviductal epithelium in vitro in relation to sperm morphology and storage time. Reproduction 131:311-318. https://doi.org/10.1530/rep.1.00814
  22. Wada K, Nomura S, Morii E, Kitamura Y, Nishizawa Y, Miyake A, Terada N. 1996. Changes in levels of mRNAs of transforming growth factor (TGF)-beta1, -beta2, -beta3, TGF-beta type II receptor and sulfated glycoprotein-2 during apoptosis of mouse uterine epithelium. J. Steroid Biochem. Mol. Biol. 59:367-375. https://doi.org/10.1016/S0960-0760(96)00139-2
  23. Wang L, Guan HT, Tian YH, Xiong CL. 2006. [Urokinase-type plasminogen activator and urokinase-type plasminogen activator receptor in the seminal plasma and sperm of fertile and oligoasthenozoospermia males]. Zhonghua Nan Ke Xue 12:791-793. Chinese.