open access

Vol 59, No 4 (2021)
Original paper
Submitted: 2021-01-07
Accepted: 2021-11-06
Published online: 2021-11-30
Get Citation

Transcriptional activation and regulation of urokinase plasminogen activator inducted by LPS through MyD88 independent pathway in rat Sertoli cells

Yan Liu12, Kai Zhao1, Zhe Xiong3, Yuanxiao Yi1, Chengliang Xiong1, Donghui Huang1, Hu Zhao4
DOI: 10.5603/FHC.a2021.0026
·
Pubmed: 34852179
·
Folia Histochem Cytobiol 2021;59(4):236-244.
Affiliations
  1. Institute of Reproduction Health, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, China
  2. Puren Hospital Affiliated to Wuhan University of Science and Technology, Wuhan, China
  3. Department of Physiology, Medical School of Jianghan University, Wuhan, China
  4. Department of Human Anatomy, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, China

open access

Vol 59, No 4 (2021)
ORIGINAL PAPERS
Submitted: 2021-01-07
Accepted: 2021-11-06
Published online: 2021-11-30

Abstract

Introduction. Urokinase plasminogen activator (uPA) is a serine protease and it also demonstrates proinflammatory properties. We thus hypothesized that uPA is involved in immunity of Sertoli cells in the rat testis.

Materials and methods. The uPA gene(PLAU) promoter was cloned by RT-PCR and the transcriptional activation of core domain was screened by using Dual-Luciferase Reporter Assay System. The Sertoli cells were harvested from 20-day-old Sprague-Dawley male rats and total RNA was isolated. The uPA mRNA levels and MyD88 pathway were tested by qPCR.

Results. We successfully cloned the 1517-bp rat uPA gene and screened the core domain (-455/+40) in five
different fragments of uPA promoter. The uPA expression and uPA promoter activity were upregulated in
lipopolysaccharide (LPS)-stimulated Sertoli cells. Furthermore, the uPA expression was regulated through the
MyD88-independent pathway by interdicting IRF3 and interferon b.

Conclusion. uPA expression is likely induced by LPS through the core promoter domain of uPA. This finding
implied that uPA played a role in the immune function of Sertoli cells in rat testis, which might provide the development of new treatments for male infertility.

Abstract

Introduction. Urokinase plasminogen activator (uPA) is a serine protease and it also demonstrates proinflammatory properties. We thus hypothesized that uPA is involved in immunity of Sertoli cells in the rat testis.

Materials and methods. The uPA gene(PLAU) promoter was cloned by RT-PCR and the transcriptional activation of core domain was screened by using Dual-Luciferase Reporter Assay System. The Sertoli cells were harvested from 20-day-old Sprague-Dawley male rats and total RNA was isolated. The uPA mRNA levels and MyD88 pathway were tested by qPCR.

Results. We successfully cloned the 1517-bp rat uPA gene and screened the core domain (-455/+40) in five
different fragments of uPA promoter. The uPA expression and uPA promoter activity were upregulated in
lipopolysaccharide (LPS)-stimulated Sertoli cells. Furthermore, the uPA expression was regulated through the
MyD88-independent pathway by interdicting IRF3 and interferon b.

Conclusion. uPA expression is likely induced by LPS through the core promoter domain of uPA. This finding
implied that uPA played a role in the immune function of Sertoli cells in rat testis, which might provide the development of new treatments for male infertility.

Get Citation

Keywords

rat; Sertoli cells; urokinase plasminogen activator; lipopolysaccharide; uPA promoter

About this article
Title

Transcriptional activation and regulation of urokinase plasminogen activator inducted by LPS through MyD88 independent pathway in rat Sertoli cells

Journal

Folia Histochemica et Cytobiologica

Issue

Vol 59, No 4 (2021)

Article type

Original paper

Pages

236-244

Published online

2021-11-30

DOI

10.5603/FHC.a2021.0026

Pubmed

34852179

Bibliographic record

Folia Histochem Cytobiol 2021;59(4):236-244.

Keywords

rat
Sertoli cells
urokinase plasminogen activator
lipopolysaccharide
uPA promoter

Authors

Yan Liu
Kai Zhao
Zhe Xiong
Yuanxiao Yi
Chengliang Xiong
Donghui Huang
Hu Zhao

References (39)
  1. Foley JH. Plasmin(ogen) at the Nexus of Fibrinolysis, Inflammation, and Complement. Semin Thromb Hemost. 2017; 43(2): 135–142.
  2. Draxler DF, Sashindranath M, Medcalf RL. Plasmin: A Modulator of Immune Function. Semin Thromb Hemost. 2017; 43(2): 143–153.
  3. Coutts SB, Dubuc V, Mandzia J, et al. TEMPO-1 Investigators. Tenecteplase-tissue-type plasminogen activator evaluation for minor ischemic stroke with proven occlusion. Stroke. 2015; 46(3): 769–774.
  4. Wang C, Huang R, Li C, et al. Vepoloxamer Enhances Fibrinolysis of tPA (Tissue-Type Plasminogen Activator) on Acute Ischemic Stroke. Stroke. 2019; 50(12): 3600–3608.
  5. Mekkawy AH, Pourgholami MH, Morris DL. Involvement of urokinase-type plasminogen activator system in cancer: an overview. Med Res Rev. 2014; 34(5): 918–956.
  6. Liu YX. Involvement of plasminogen activator and plasminogen activator inhibitor type 1 in spermatogenesis, sperm capacitation, and fertilization. Semin Thromb Hemost. 2007; 33(1): 29–40.
  7. Martinez-Soto JC, Landeras J, Mollá M, et al. Total urokinase-type plasminogen activator (uPA) levels in seminal plasma are associated with positive assisted reproductive technology outcomes. J Assist Reprod Genet. 2018; 35(6): 1091–1101.
  8. Penttilä TL, Kaipia A, Toppari J, et al. Localization of urokinase- and tissue-type plasminogen activator mRNAs in rat testes. Mol Cell Endocrinol. 1994; 105(1): 55–64.
  9. Zhang T, Zhou HM, Liu YX. Expression of plasminogen activator and inhibitor, urokinase receptor and inhibin subunits in rhesus monkey testes. Mol Hum Reprod. 1997; 3(3): 223–231.
  10. Huang DH, Zhao Hu, Tian YH, et al. Gene expression changes of urokinase plasminogen activator and urokinase receptor in rat testes at postnatal stages. Asian J Androl. 2007; 9(5): 679–683.
  11. Zhao K, Liu Y, Xiong Z, et al. Tissue-specific inhibition of urokinase-type plasminogen activator expression in the testes of mice by inducible lentiviral RNA interference causes male infertility. Reprod Fertil Dev. 2017; 29(11): 2149–2156.
  12. Foley JH. Plasmin(ogen) at the Nexus of Fibrinolysis, Inflammation, and Complement. Semin Thromb Hemost. 2017; 43(2): 135–142.
  13. Draxler DF, Sashindranath M, Medcalf RL. Plasmin: A Modulator of Immune Function. Semin Thromb Hemost. 2017; 43(2): 143–153.
  14. Ding X, Li H, Li Y, et al. Two B-cell epitope vaccines based on uPA effectively inhibit fertility in male mice. Vaccine. 2018; 36(19): 2612–2618.
  15. Li X, Egervari G, Wang Y, et al. Regulation of chromatin and gene expression by metabolic enzymes and metabolites. Nat Rev Mol Cell Biol. 2018; 19(9): 563–578.
  16. Vernimmen D, Bickmore WA. The Hierarchy of Transcriptional Activation: From Enhancer to Promoter. Trends Genet. 2015; 31(12): 696–708.
  17. LIU Y, XIONG Jw, CHEN Lg, et al. Cloning and Analysis of the Promoter Region of Rat uPA Gene. Journal of Reproduction and Contraception. 2007; 18(1): 1–10.
  18. Haile WB, Coleman JL, Benach JL. Reciprocal upregulation of urokinase plasminogen activator and its inhibitor, PAI-2, by Borrelia burgdorferi affects bacterial penetration and host-inflammatory response. Cell Microbiol. 2006; 8(8): 1349–1360.
  19. Hoenigl M, Raggam RB, Wagner J, et al. Diagnostic accuracy of soluble urokinase plasminogen activator receptor (suPAR) for prediction of bacteremia in patients with systemic inflammatory response syndrome. Clin Biochem. 2013; 46(3): 225–229.
  20. Yeh YH, Wang PH, Lin LY, et al. Significantly increased concentration of soluble urokinase-type plasminogen activator receptor in the blood of patients with pelvic inflammatory disease. Clin Chim Acta. 2013; 415: 138–144.
  21. Xia Y, Yuan M, Li S, et al. Apigenin Suppresses the IL-1β-Induced Expression of the Urokinase-Type Plasminogen Activator Receptor by Inhibiting MAPK-Mediated AP-1 and NF-κB Signaling in Human Bladder Cancer T24 Cells. J Agric Food Chem. 2018; 66(29): 7663–7673.
  22. Firmal P, Shah VK, Chattopadhyay S. Insight Into TLR4-Mediated Immunomodulation in Normal Pregnancy and Related Disorders. Front Immunol. 2020; 11: 807.
  23. Clifton RJ, O'Donnell L, Robertson DM. Pachytene spermatocytes in co-culture inhibit rat Sertoli cell synthesis of inhibin beta B-subunit and inhibin B but not the inhibin alpha-subunit. J Endocrinol. 2002; 172(3): 565–574.
  24. Winnall WR, Muir JA, Hedger MP. Differential responses of epithelial Sertoli cells of the rat testis to Toll-like receptor 2 and 4 ligands: implications for studies of testicular inflammation using bacterial lipopolysaccharides. Innate Immun. 2011; 17(2): 123–136.
  25. Moquet-Torcy G, Tolza C, Piechaczyk M, et al. Transcriptional complexity and roles of Fra-1/AP-1 at the uPA/Plau locus in aggressive breast cancer. Nucleic Acids Res. 2014; 42(17): 11011–11024.
  26. Lu KH, Yang HW, Su CW, et al. Phyllanthus urinaria suppresses human osteosarcoma cell invasion and migration by transcriptionally inhibiting u-PA via ERK and Akt signaling pathways. Food Chem Toxicol. 2013; 52: 193–199.
  27. Alcaraz-Pérez F, Mulero V, Cayuela ML. Application of the dual-luciferase reporter assay to the analysis of promoter activity in Zebrafish embryos. BMC Biotechnol. 2008; 8: 81.
  28. Lie PPY, Cheng CY, Mruk DD. Signalling pathways regulating the blood-testis barrier. Int J Biochem Cell Biol. 2013; 45(3): 621–625.
  29. Zhao S, Zhu W, Xue S, et al. Testicular defense systems: immune privilege and innate immunity. Cell Mol Immunol. 2014; 11(5): 428–437.
  30. Li N, Wang T, Han D. Structural, cellular and molecular aspects of immune privilege in the testis. Front Immunol. 2012; 3: 152.
  31. Slomiany BL, Slomiany A. Role of LPS-elicited signaling in triggering gastric mucosal inflammatory responses to H. pylori: modulatory effect of ghrelin. Inflammopharmacology. 2017; 25(4): 415–429.
  32. Michailidis G, Anastasiadou M, Guibert E, et al. Activation of innate immune system in response to lipopolysaccharide in chicken Sertoli cells. Reproduction. 2014; 148(3): 259–270.
  33. Yeagley D, Lang CH. Endotoxin-Induced IL-6 Promoter Activation in Skeletal Muscle Requires an NF-κB Site. Int J Interferon Cytokine Mediat Res. 2010; 2010(2): 9–21.
  34. Casals-Casas C, Alvarez E, Serra M, et al. CREB and AP-1 activation regulates MKP-1 induction by LPS or M-CSF and their kinetics correlate with macrophage activation versus proliferation. Eur J Immunol. 2009; 39(7): 1902–1913.
  35. Rocha DM, Caldas AP, Oliveira LL, et al. Saturated fatty acids trigger TLR4-mediated inflammatory response. Atherosclerosis. 2016; 244: 211–215.
  36. Kawai T, Takeuchi O, Fujita T, et al. Lipopolysaccharide stimulates the MyD88-independent pathway and results in activation of IFN-regulatory factor 3 and the expression of a subset of lipopolysaccharide-inducible genes. J Immunol. 2001; 167(10): 5887–5894.
  37. Zhang H, Yin Y, Wang G, et al. Interleukin-6 disrupts blood-testis barrier through inhibiting protein degradation or activating phosphorylated ERK in Sertoli cells. Sci Rep. 2014; 4: 4260.
  38. Lei T, Moos S, Klug J, et al. Galectin-1 enhances TNFα-induced inflammatory responses in Sertoli cells through activation of MAPK signalling. Sci Rep. 2018; 8(1): 3741.
  39. Iliev DB, Sobhkhez M, Fremmerlid K, et al. MyD88 interacts with interferon regulatory factor (IRF) 3 and IRF7 in Atlantic salmon (Salmo salar): transgenic SsMyD88 modulates the IRF-induced type I interferon response and accumulates in aggresomes. J Biol Chem. 2011; 286(49): 42715–42724.

Regulations

Important: This website uses cookies. More >>

The cookies allow us to identify your computer and find out details about your last visit. They remembering whether you've visited the site before, so that you remain logged in - or to help us work out how many new website visitors we get each month. Most internet browsers accept cookies automatically, but you can change the settings of your browser to erase cookies or prevent automatic acceptance if you prefer.

By "Via Medica sp. z o.o." sp.k., ul. Świętokrzyska 73, 80–180 Gdańsk

tel.:+48 58 320 94 94, faks:+48 58 320 94 60, e-mail:  viamedica@viamedica.pl