Original Article

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



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.


Animal ethics
Isolation and culture of pUECs and uterine tissue epithelial cell
Preparation and co-incubation of boar sperm
cDNA synthesis and real-time PCR
Measurement of PA activity
Flow cytometry (FACs) method of sperm
Statistical analysis
The role of sperm in porcine uterine tissue epithelial cells
Effect of PA on sperm survival rate and acrosome


  • Su-jin Kim College of Animal Life Sciences, Kangwon National University, Chuncheon 24341, Korea
  • Hee-Tae Cheong College of Veterinary Medicine, Kangwon National University, Chuncheon 24341, Korea
  • Choon-keun Park College of Animal Life Sciences, Kangwon National University, Chuncheon 24341, Korea


자료제공 : 네이버학술정보

    함께 이용한 논문

      ※ 기관로그인 시 무료 이용이 가능합니다.

      • 4,000원

      0개의 논문이 장바구니에 담겼습니다.