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Effects of Progesterone and 17β-Estradiol under Presence or Absence of FBS on Plasminogen Activators Activity in Porcine Uterine Epithelial Cells

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초록

The present study was conducted to investigate the regulatory mechanism of plasminogen activators (PAs) ac-tivation by 17β-estradiol (E2) and progesterone (P4) in porcine uterine epithelial cells (pUECs). pUECs were collected from porcine uterine horn and cultured at 80% confluence. Then, 0.1% (v/v) DMSO, 20 ng/mL E2, and P4 with or without fetal bo-vine serum (FBS) treated to cultured cells for 24 hours. The supernatants were used for measurement of PAs activity and ex-pression of urokinase-type PA (uPA), tissue-type PA (tPA), uPA specific receptor (uPAR), and type-1 PA inhibitor (PAI-1) mRNA were analyzed by real-time PCR. The expression of PAs-related genes was not affect by steroid hormones in both of serum treatment groups. However, PAs activity was increased by treatment of E2 compared to 0.1% DMSO treatment in se-rum-free group (p<0.05). Then, E2 and P4 were diluted with 0.002% (v/v) DMSO for reduction of its effect and treated to cul-tured cells without FBS. Only tPA mRNA was significantly increased by E2 treatment (p<0.05). PAs activity was enhanced in E2 treated group compared to control groups (p<0.05). These results indicate that serum-free condition is more proper to eval-uate effect of steroid hormones and activation of PAs in pUECs was mainly regulated by estrogen. These regulation of PAs activation may be associated with uterine remodeling during pre-ovulatory phase in pigs, however, further studies are needed to investigate precise regulatory mechanism.

Abstract

The present study was conducted to investigate the regulatory mechanism of plasminogen activators (PAs) ac-tivation by 17β-estradiol (E2) and progesterone (P4) in porcine uterine epithelial cells (pUECs). pUECs were collected from porcine uterine horn and cultured at 80% confluence. Then, 0.1% (v/v) DMSO, 20 ng/mL E2, and P4 with or without fetal bo-vine serum (FBS) treated to cultured cells for 24 hours. The supernatants were used for measurement of PAs activity and ex-pression of urokinase-type PA (uPA), tissue-type PA (tPA), uPA specific receptor (uPAR), and type-1 PA inhibitor (PAI-1) mRNA were analyzed by real-time PCR. The expression of PAs-related genes was not affect by steroid hormones in both of serum treatment groups. However, PAs activity was increased by treatment of E2 compared to 0.1% DMSO treatment in se-rum-free group (p<0.05). Then, E2 and P4 were diluted with 0.002% (v/v) DMSO for reduction of its effect and treated to cul-tured cells without FBS. Only tPA mRNA was significantly increased by E2 treatment (p<0.05). PAs activity was enhanced in E2 treated group compared to control groups (p<0.05). These results indicate that serum-free condition is more proper to eval-uate effect of steroid hormones and activation of PAs in pUECs was mainly regulated by estrogen. These regulation of PAs activation may be associated with uterine remodeling during pre-ovulatory phase in pigs, however, further studies are needed to investigate precise regulatory mechanism.

참고문헌 (41)

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